阅读说明：本技术 一种用于治疗女性阴道松弛及压力性尿失禁的冷敷凝胶及其制备方法 (Cold compress gel for treating female vaginal relaxation and stress urinary incontinence and preparation method thereof ) 是由 陈海云 任丽琴 林俊锦 于 2021-08-18 设计创作，主要内容包括：本发明公开了一种用于治疗女性阴道松弛及压力性尿失禁的冷敷凝胶及其制备方法。该冷敷凝胶包括以下有效成分：莪术提取物、补骨脂、铁皮石斛水提物、大豆异黄酮、乳香酸、水解交联胶原蛋白、纤维粘连蛋白。本发明中各组份合理搭配,制备过程极大保留有效成分,有效成分直接相互协同相互促进,通过阴道给药后的黏膜吸收,促进盆底肌肉组织的收缩,修复肌神经,提升肌肉的张力,实现盆底功能障碍的治疗,对改善因盆底损伤导致的压力性尿失禁具有显著的效果。(The invention discloses a cold compress gel for treating female vaginal relaxation and stress urinary incontinence and a preparation method thereof. The cold compress gel comprises the following effective components: zedoary turmeric extract, fructus Psoraleae, herba Dendrobii water extract, soybean isoflavone, Olibanum acid, hydrolyzed crosslinked collagen, and fibronectin. The components of the invention are reasonably matched, the preparation process greatly retains the effective components, the effective components are directly mutually cooperated and mutually promoted, and the contraction of pelvic floor muscle tissues is promoted, the muscular nerve is repaired, the tension of muscles is promoted through the mucous membrane absorption after the vaginal administration, the treatment of the pelvic floor dysfunction is realized, and the invention has obvious effect on improving the stress urinary incontinence caused by the pelvic floor injury.)

1. A cold compress gel for treating female vaginal relaxation and stress urinary incontinence, comprising the following active ingredients: zedoary turmeric extract, fructus Psoraleae, herba Dendrobii water extract, soybean isoflavone, Olibanum acid, hydrolyzed crosslinked collagen, and fibronectin.

2. A cold compress gel according to claim 1, comprising the following active ingredients: 0.3 to 1.6 percent of zedoary extract, 0.5 to 2 percent of malaytea scurfpea fruit, 0.5 to 2 percent of dendrobium officinale aqueous extract, 0.03 to 0.6 percent of soybean isoflavone, 0.03 to 0.6 percent of boswellic acid, 0.08 to 1.2 percent of hydrolytic cross-linked collagen and 0.01 to 0.15 percent of fibronectin.

3. A cold compress gel according to claim 1 or 2, further comprising the following adjuvants: 0.1-1% of carbomer, 1-10% of glycerol, 1-10% of propylene glycol, 0.04-0.4% of methyl paraben, 0.04-0.4% of propyl paraben, 0.15-1.5% of triethanolamine and the balance of deionized water to 100%.

4. A method of preparing a cold compress gel for treating female vaginal relaxation and stress urinary incontinence as claimed in claim 3, comprising the steps of:

(1) weighing corresponding mass of Curcumae rhizoma extract, fructus Psoraleae, herba Dendrobii hydrolysate, Olibanum acid and soybean isoflavone, and uniformly dispersing the raw materials with propylene glycol to obtain mixture A;

(2) weighing carbomer powder and hydrolyzed collagen with corresponding mass, adding into deionized water, adding glycerol, heating to 60-90 deg.C, stirring thoroughly to dissolve to obtain mixture B;

(3) when the temperature of the mixture B is reduced to 35-50 ℃, adding fibronectin, methyl paraben and propyl paraben while stirring to obtain a mixture C;

(4) a, C is mixed and stirred evenly at the temperature of 35-50 ℃, and the mixture D is obtained after homogenizing for 3-6min and 1000-1500 r/min;

(5) and adding triethanolamine into the mixture D, and fully and uniformly stirring to obtain the cold compress gel.

5. The method for preparing a cold compress gel for treating female vaginal relaxation and stress urinary incontinence as claimed in claim 4, wherein the extraction process of the zedoary turmeric extract comprises: zedoary turmeric oil is used as raw material, silica gel column chromatography is carried out, petroleum ether is used for elution, ethyl acetate elution part is collected, and brown yellow oily matter obtained after ethyl acetate is recovered in vacuum is zedoary turmeric extract.

6. The method for preparing the cold compress gel for treating female vaginal relaxation and stress urinary incontinence as claimed in claim 4, wherein the extraction process of said dendrobium water extract is: firstly, crushing the dendrobium officinale by using a crusher, leaching the dendrobium officinale by hot water at the temperature of 80 +/-10 ℃ for 1 hour according to the material-liquid ratio of 1: 30, filtering by using double-layer medical gauze, retaining filter residues, repeatedly leaching by hot water for 3 times according to the material-liquid ratio of 1: 30, 1: 20 and 1: 10, combining all filtrates, centrifuging, and taking supernatant; concentrating under reduced pressure to 1/6-1/8 volume of the original solution, and freezing at-20 deg.C in refrigerator; and (3) carrying out freeze drying on the frozen concentrated solution by using a vacuum freeze dryer to obtain a dried dendrobium officinale extract with the water content of less than 2.5%.

Technical Field

The invention relates to the technical field of medicine preparation, in particular to cold compress gel for treating female vaginal relaxation and stress urinary incontinence and a preparation method thereof.

Background

Female pelvic floor dysfunctional disease (PFD) is a common gynecological disease, is clinically manifested as Stress Urinary Incontinence (SUI), vaginal relaxation (VL), Pelvic Organ Prolapse (POP), Female Sexual Dysfunction (FSD) and the like, and seriously affects the health and the quality of life of middle-aged and old women. The incidence of SUI in China is 22.9%. The relevant united states authorities represent a recent incidence of SUI of up to 25% with patients presenting with POP and big, urinary incontinence in the order of 2.9%, 9.4% and 17.1%. PFD has recently become a concern for this disease in most countries around the world, where female health has been an indispensable and hard indicator for assessing public health levels, and where prevention of female PFD has been one of the major government tasks.

The most common symptoms of pelvic floor dysfunction are vaginal relaxation and stress urinary incontinence, and the existing treatment technology comprises three schemes of conservative treatment, drug treatment and surgical treatment.

(1) The conservative treatment is pelvic floor muscle training treatment and comprises the following three methods:

the method comprises the following steps of: by autonomous and repeated contraction and relaxation of pelvic floor muscle groups, the tension of pelvic floor muscles supporting urethra, bladder, uterus and rectum is enhanced, the resistance of urethra is increased, the function of pelvic floor muscles is recovered, and the purpose of preventing and treating urinary incontinence is achieved. Pelvic floor muscle training must be performed with regular training and long persistence before it can be effective.

Bioelectrical feedback: by means of the electronic biofeedback therapeutic equipment placed in vagina or rectum, the myoelectric activity of pelvic floor muscles is monitored, a patient is guided to carry out correct and independent pelvic floor muscle training, conditioned reflex is formed, and the curative effect is equivalent to that of pure pelvic floor muscle training.

(iii) electrical stimulation treatment: the current repeatedly stimulates pelvic floor nerves and muscles, increases the contractility of the pelvic floor muscles, feedbacks and inhibits sympathetic nerve reflex, and reduces the activity of the bladder. But cardiac pacemaker, pregnancy, severe pelvic organ prolapse, vaginitis and hemorrhage are contraindications, and the method cannot be adopted.

(2) The main action principle of the drug therapy is to increase the urethral closing pressure, and the current commonly used drugs comprise the following drugs:

selective alpha l-adrenoceptor agonists: is effective for SUI, especially when estrogen or pelvic floor muscle training is used simultaneously. The common medicines are as follows: midodrine hydrochloride tablets. The side effects include hypertension, palpitation, headache, cold extremities, and severe apoplexy.

② estrogen: promoting hyperplasia of urethral mucosa, submucosal vascular plexus and connective tissue; enhance the number and sensitivity of alpha adrenergic receptors. Oral administration of estrogen does not reduce urinary incontinence and risks inducing and aggravating urinary incontinence. Topical vaginal estrogen is beneficial for SUI.

(3) The operation treatment mainly aims at the patients who need pelvic floor reconstruction operation for POP accompanied with SUI which seriously affects the life quality, and anti-SUI operation should be carried out at the same time. The main surgical methods are:

tension-free mid-urethral suspension, posterior cupressus ligamentum suspension (Burch). Complications include: bladder perforation, bleeding, difficulty in urination, rejection or delayed healing of an inserted sling, erosion of the human urethra or vagina by the sling, intestinal perforation, infection, and the like.

However, the above methods all have certain defects, specifically:

autonomous pelvic floor muscle exercise: the pelvic floor muscle function recovery through exercise requires long-term adherence and has certain effect after variable accumulation. Therefore, this method is long in time, poor in compliance, and not ideal in effect.

Bioelectric feedback, electrical muscle stimulation: pelvic floor dysfunction is a long-term rehabilitation process, and the method needs to be treated in medical institutions, and has poor compliance and undesirable effects.

And (3) drug treatment: currently, the only drugs clinically used to treat this disease are class 2: the alpha l-adrenoceptor agonist and the estrogen have few optional medicines and large side effect, and a single action mechanism improves the function of pelvic floor muscles, so the effect is not ideal.

And (3) surgical treatment: firstly, the operation wound is large; secondly, the tightness is not easy to master, and the technical requirement is high; high complication rate; fourthly, the operation time is long; fifthly, the cost is high.

Therefore, the problem to be solved by the technical personnel in the field is how to provide a medicine for treating female vaginal relaxation and stress urinary incontinence, which has good effect, no side effect and convenient implementation.

Disclosure of Invention

In view of the above, the invention provides a cold compress gel for treating female vaginal relaxation and stress urinary incontinence and a preparation method thereof, the components of the cold compress gel are reasonably matched, the preparation process greatly retains effective components, the effective components are directly mutually cooperated and mutually promoted, the contraction of pelvic floor muscle tissues is promoted, the muscular nerve is repaired, the tension of muscles is improved through the mucous membrane absorption after vaginal administration, the treatment of pelvic floor dysfunction is realized, and the cold compress gel has a remarkable effect on improving the stress urinary incontinence caused by pelvic floor injury.

In order to achieve the purpose, the invention adopts the following technical scheme:

a cold compress gel for treating female vaginal relaxation and stress urinary incontinence comprises the following effective components: zedoary turmeric extract, fructus Psoraleae, herba Dendrobii water extract, soybean isoflavone, Olibanum acid, hydrolyzed crosslinked collagen, and fibronectin.

Adopt above-mentioned technical scheme's beneficial effect:

the main reason for pelvic floor muscle relaxation is that fibroblasts are rapidly apoptotic under the influence of mechanical force, so that the number of the fibroblasts is continuously reduced, related collagen, enzyme, EGFR growth factor, estrogen and the like are secreted, and the structure and function of supporting tissues are greatly weakened. The cold compress gel of the invention comprises the following main components: zedoary turmeric extract, fructus Psoraleae, herba Dendrobii water extract, soybean isoflavone, Olibanum acid, hydrolyzed crosslinked collagen, and fibronectin. The technical key points are that the pelvic floor muscle function is repaired from the following aspects aiming at the relaxation of the pelvic floor muscle:

(1) promoting the division and proliferation of fibroblasts, accelerating the secretion of self collagen, and restoring the elasticity of the pelvic floor muscle so as to restore the tissue structure and the function of the pelvic floor muscle;

(2) the expression of EGFR protein is effectively regulated, so that the C2C12 cell proliferation promoting effect is achieved, and the tension-lost pelvic floor muscle group is repaired;

(3) the nerve injury is protected and repaired by promoting the secretion of factors such as NGF/BDNF of Schwann cells and glial cells;

(4) promote the contraction of the vaginal wall and enter the pelvic floor microcirculation through transdermal absorption to ensure the contraction of pelvic floor muscles;

(5) the invention adopts phytoestrogen to replace estrogen: as an important hormone, estrogen has wide and important physiological effects, and can be used for participating in the occurrence and development of various diseases and also can be used as a means for treating the diseases. However, the estrogen is just like a double-edged sword, and when the estrogen is selected as a therapeutic drug, human beings have to bear a large amount of adverse reactions brought by the estrogen. Therefore, phytoestrogens which have estrogenic effects and do not cause serious adverse reactions bring new hopes for the treatment of diseases. The active ingredients extracted from the traditional Chinese medicine for supplementing the bone fat, such as psoralen, soybean isoflavone and the like, have estrogen-like effect, have less obvious effect than estrogen in pelvic floor muscle repair and stress urinary incontinence improvement, but have good prospect when being used as a treatment medicine for mild symptoms or being combined with other medicines.

The cold compress gel of the invention has the following action principle:

(1) the zedoary turmeric extract has the functions of promoting the contraction of the vaginal wall and contracting the pelvic floor muscles through percutaneous absorption and entering the pelvic floor microcirculation, and is beneficial to recovering the functions of the elastic supporting structure of the vaginal wall and the pelvic floor muscles. The zedoary turmeric extract has alpha adrenergic receptor stimulant function, can promote the contraction of vaginal wall and pelvic floor muscle, and achieves the effect of improving the functions of the vagina and the pelvic floor muscle. Meanwhile, the zedoary turmeric extract has M choline receptor agonism function, and further strengthens the contraction effect of vagina and pelvic floor muscles.

(2) Fructus Psoraleae contains various active components, such as coumarin-like psoralen, isobavachin, fructus Psoraleae tincture, flavonoid component psoralen B, and monopegenin component bakuchiol, etc., and has estrogen-like effect. Radix astragali contains radix astragali, rhizoma corydalis, B-sitosterol, etc., and has effects of promoting estrogen. It can promote pituitary-gonad function and increase organic estrogen level. Thereby promoting the repair of vagina and urethra mucous membrane, improving the internal contracting function of the vagina and urethra, increasing the vaginal and urethra closing force and achieving the effects of vagina contraction and urinary incontinence.

(3) The dendrobium officinale aqueous extract can effectively regulate the expression of EGFR protein, thereby playing a role in promoting the proliferation of C2C12 cells. By utilizing the pelvic floor skeletal muscle tissue in-vitro incubation experiment system, the pelvic floor skeletal muscle tissue is incubated by using a buffer solution containing a dendrobium officinale aqueous extract. Through experiments, the EGFR expression level of skeletal muscle tissues treated by the dendrobium officinale aqueous extract is remarkably increased compared with that of a control group. The herba Dendrobii extract enters microcirculation through transdermal absorption and acts on pelvic floor muscle group, and has repairing effect on pelvic floor muscle group with tensile loss.

(4) Soy isoflavone (soyaisoflavanes. sif) is a natural Phytoestrogen (Phytoestrogen PE). The soybean isoflavone has a similar molecular structure with estrogen, and competes with estrogen in women to bind to estrogen receptor, so that the estrogen level in the body is kept balanced. When women over the age of 35 years experience a decline in estrogen levels in the body and adolescent activity is affected. And proper amount of exogenous soybean isoflavone can improve the symptoms of vaginal and pelvic floor muscle relaxation and the like.

(5) The boswellic acid in the frankincense has a nerve protection effect, and the nerve protection effect is realized by secreting neurotrophic factor G through thorn number cells; olibanum has the effect of repairing nerve injury by promoting axon growth.

(6) The fibronectin and the hydrolyzed collagen are extracellular matrix components and exist in interstitial substance of cells, and the fibronectin and the collagen are used together, so that the myocytes can be promoted to secrete collagen and other extracellular matrix proteins, the elasticity of vagina and pelvic floor muscles is restored, and the tissue structure and the function of the vagina and the pelvic floor muscles are restored.

The above components can play a synergistic role, such as: the sensitivity of alpha adrenergic receptor can be further improved by the action of the soybean isoflavone estrogen-like substance, and the stimulation of the zedoary turmeric extract on the contraction of vagina and pelvic floor muscles can be enhanced. The dendrobium officinale aqueous extract can effectively regulate the expression of EGFR protein and further promote muscle cells to secrete extracellular matrix proteins such as collagen and the like. The fructus psoraleae contains various active ingredients and soybean isoflavone, and can play a synergistic effect and the like for different types of phytoestrogens.

Preferably, the cold compress gel comprises the following effective components: 0.3 to 1.6 percent of zedoary extract, 0.5 to 2 percent of malaytea scurfpea fruit, 0.5 to 2 percent of dendrobium officinale aqueous extract, 0.03 to 0.6 percent of soybean isoflavone, 0.03 to 0.6 percent of boswellic acid, 0.08 to 1.2 percent of hydrolytic cross-linked collagen and 0.01 to 0.15 percent of fibronectin.

Preferably, the cold compress gel further comprises the following auxiliary materials: 0.1-1% of carbomer, 1-10% of glycerol, 1-10% of propylene glycol, 0.04-0.4% of methyl paraben, 0.04-0.4% of propyl paraben, 0.15-1.5% of triethanolamine and the balance of deionized water to 100%.

The invention adopts propylene glycol as a penetration enhancer. Penetration enhancers refer to chemical substances that reversibly alter the barrier function of the stratum corneum of the skin without damaging any active cells. The ideal penetration enhancer should have the following characteristics: the composition has chemical inertia, chemical stability and no pharmacological activity; ② reversibly change the skin characteristics, the onset is fast; thirdly, no incompatibility exists between the medicine and the matrix; fourthly, the medicine is non-toxic, non-irritant, non-allergic and non-allergic; no color, no smell and no smell; sixthly, the skin-care product is easy to spread on the skin, has no discomfort and has good compatibility with the skin. The present inventors have also studied the release of propylene glycol, a penetration enhancer in a transdermal drug delivery system, and have found that the release of the penetration enhancer conforms to the Higuchi formula and that the release rate is inversely proportional to the cubic root of the molar volume of the penetration enhancer and the logarithm of the partition coefficient between the matrix and the receiving liquid, the kind of matrix has less influence on the release rate, and it is most preferable to use propylene glycol as the penetration enhancer of the present invention.

The invention also provides a preparation method of the cold compress gel for treating female vaginal relaxation and stress urinary incontinence, which comprises the following steps:

(1) weighing corresponding mass of Curcumae rhizoma extract, fructus Psoraleae, herba Dendrobii hydrolysate, Olibanum acid and soybean isoflavone, and uniformly dispersing the raw materials with propylene glycol to obtain mixture A;

(2) weighing carbomer powder and hydrolyzed collagen with corresponding mass, adding into deionized water, adding glycerol, heating to 60-90 deg.C, stirring thoroughly to dissolve to obtain mixture B;

(3) when the temperature of the mixture B is reduced to 35-50 ℃, adding fibronectin, methyl paraben and propyl paraben while stirring to obtain a mixture C;

(4) a, C is mixed and stirred evenly at the temperature of 35-50 ℃, and the mixture D is obtained after homogenizing for 3-6min and 1000-1500 r/min;

(5) and adding triethanolamine into the mixture D, and fully and uniformly stirring to obtain the cold compress gel.

Preferably, the extraction process of the curcuma zedoary extract comprises the following steps: zedoary turmeric oil is used as raw material, silica gel column chromatography is carried out, petroleum ether is used for elution, ethyl acetate elution part is collected, and brown yellow oily matter obtained after ethyl acetate is recovered in vacuum is zedoary turmeric extract.

Preferably, the extraction process of the dendrobium water extract comprises the following steps: firstly, crushing the dendrobium officinale by using a crusher, leaching the dendrobium officinale by hot water at the temperature of 80 +/-10 ℃ for 1 hour according to the material-liquid ratio of 1: 30, filtering by using double-layer medical gauze, retaining filter residues, repeatedly leaching by hot water for 3 times according to the material-liquid ratio of 1: 30, 1: 20 and 1: 10, combining all filtrates, centrifuging, and taking supernatant; concentrating under reduced pressure to 1/6-1/8 volume of the original solution, and freezing at-20 deg.C in refrigerator; and (3) carrying out freeze drying on the frozen concentrated solution by using a vacuum freeze dryer to obtain a dried dendrobium officinale extract with the water content of less than 2.5%.

Compared with the prior art, the invention solves the following problems:

(1) the optimization of phytoestrogen solves the problem of side effect of the existing clinical common medicines. The invention adopts phytoestrogen to replace estrogen, and the medication route is vaginal administration and transdermal absorption through vaginal mucosa. Estrogens, an important hormone, have a wide range of important physiological effects. It can be used for the development of various diseases and as a means for treating the diseases. However, the estrogen is just like a double-edged sword, and when the estrogen is selected as a therapeutic drug, human beings have to bear a large amount of adverse reactions brought by the estrogen. Therefore, phytoestrogens which have estrogenic effects and do not cause serious adverse reactions bring new hopes for the treatment of diseases. The active ingredients extracted from the traditional Chinese medicine for supplementing the bone fat, such as psoralen, soybean isoflavone and the like, have estrogen-like effect, have less obvious effect than estrogen in pelvic floor muscle repair and stress urinary incontinence improvement, but have good prospect when being used as a treatment medicine for mild symptoms or being combined with other medicines.

(2) The preferred penetration enhancers are those that reversibly alter the barrier function of the stratum corneum of the skin without damaging any active cellular chemicals: the composition has chemical inertia, chemical stability and no pharmacological activity; ② reversibly change the skin characteristics, the onset is fast; thirdly, no incompatibility exists between the medicine and the matrix; fourthly, the medicine is non-toxic, non-irritant, non-allergic and non-allergic; no color, no smell and no smell; sixthly, the skin-care product is easy to spread on the skin, has no discomfort and has good compatibility with the skin.

(3) Solves the problem that the curative effect of the current clinical commonly used medicines is not ideal: selective α l adrenoceptor agonists: by activating the smooth muscle alpha l adrenergic receptor of urethra and the motor neuron of body, the resistance of urethra is increased, and the effective rate is about 30%. ② estrogen: promoting hyperplasia of urethral mucosa, submucosal vascular plexus and connective tissue; enhance the number and sensitivity of alpha adrenergic receptors. Oral administration of estrogen does not reduce urinary incontinence and risks inducing and aggravating urinary incontinence.

(4) The compliance problem is solved: firstly, the treatment mode of pelvic floor muscle training can be effective only when a considerable amount of training is achieved, and the defects of poor compliance and difficulty in mastering training skills exist; ② the clinical commonly used medicament alpha 1 adrenoceptor agonist and estrogen have larger adverse reaction, and the compliance of patients is not high after long-term mass use is not recommended. The cold compress gel is brought back to home under the guidance of doctors, and has the advantages of simple and convenient operation, safety, effectiveness and high compliance.

Compared with the prior art, the invention has the following advantages:

(1) different treatment modes and ways can achieve better treatment effect by combining multiple modes clinically. The cold compress gel administration route is vaginal injection administration, and can be used for combined treatment with other treatment modes such as pelvic rehabilitation training, muscle electrical stimulation, biological point feedback and the like, so that a better treatment effect is achieved.

(2) Strong transdermal absorption and improved therapeutic effect. Propylene glycol as a penetration enhancer can achieve ideal penetration characteristics: the composition has chemical inertia, chemical stability and no pharmacological activity; ② reversibly change the skin characteristics, the onset is fast; thirdly, no incompatibility exists between the medicine and the matrix; fourthly, the medicine is non-toxic, non-irritant, non-allergic and non-allergic; no color, no smell and no smell; sixthly, the skin-care product is easy to spread on the skin, has no discomfort and has good compatibility with the skin.

(3) Progress from symptomatic treatment to causal treatment: through absorption of vaginal mucosa, collagen and estrogen are supplemented exogenously, and secretion of tissue EGFR growth factor and estrogen is promoted to promote and support tissue structure and function, so that pelvic floor muscle tissue is repaired, and pelvic floor dysfunction diseases such as stress urinary incontinence and the like are fundamentally treated.

(4) The cold compress gel is injected into vagina for administration, can be brought back to home under the guidance of doctors in professional medical institutions, and is safe, effective and high in compliance.

(5) The cold compress gel adopts the plant estrogen which has estrogen-like effect, has no adverse reaction and high safety, and has the same effect as the cold compress gel which replaces the estrogen for treatment.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the provided drawings without creative efforts.

FIG. 1 is a graph of scratch test conducted on different groups of treated C2C12 myoblasts;

FIG. 2 is a graph showing the expression level of EGFR in C2C12 cells after 2h treatment of different groups; p <0.05, P <0.01 compared to placebo;

FIG. 3 shows the mobility of C2C12 cells after 2.5h of treatment for different groups; p <0.05 compared to placebo;

FIG. 4 shows Western blot to detect the expression of TGF-beta 1, Smad2/3, p-Smad2/3 proteins in the vaginal tissues of rats in each group;

in fig. 1:

a is scratch of a blank control group of 0 h; b is 2h blank control group scratch mark; c is 0h cold compress gel (1g) scratch; d is 2h cold compress gel (1g) scratch; e is 0h cold compress gel (3.5g) scratch; f is the scratch of cold compress gel (3.5g) for 2 h; g is 0h cold compress gel (7G) scratch; h is 2H cold compress gel (7g) scratch;

in fig. 4:

a is a Western blot detection result; b is semi-quantitative analysis;

FIG. A, 1, shows a sham group; 2 is a model group; 3 is a collagen group; and 4 is an estrogen group.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1

Weighing 1g of curcuma zedoary extract, 1g of fructus psoraleae, 1g of dendrobium officinale hydrolysate, 0.4g of boswellic acid and 0.4g of soybean isoflavone, and uniformly dispersing the five raw materials by using 8g of propylene glycol to obtain a mixture A;

weighing 0.5g of carbomer U20 powder and 0.4g of hydrolyzed collagen, adding into 100g of deionized water, adding 6g of glycerol, heating to 80 ℃, and fully stirring for dissolving to obtain a mixture B;

when the temperature of the mixture B is reduced to 45 ℃, adding 0.1g of fibrous adhesion egg yolk, 0.13g of methyl hydroxybenzoate and 0.13g of propyl hydroxybenzoate while stirring to obtain a mixture C;

mixing and stirring the mixture A, C uniformly at 45 ℃, homogenizing for 5min at the rotation speed of 1500 revolutions per minute to obtain a mixture D;

and adding triethanolamine into the mixture D, stirring uniformly, filling the gel final product into a vaginal push tube, and filling 3.5g of gel into each tube to obtain the cold compress gel which can effectively treat stress urinary incontinence caused by pelvic floor muscle relaxation.

Example 2

Weighing 0.3g of curcuma zedoary extract, 0.5g of fructus psoraleae, 0.5g of dendrobium officinale hydrolysate, 0.03g of boswellic acid and 0.03g of soybean isoflavone, and uniformly dispersing the five raw materials by using 1g of propylene glycol to obtain a mixture A;

weighing 0.1g of carbomer U20 powder and 0.1g of hydrolyzed collagen, adding into 70g of deionized water, adding 1g of glycerol, heating to 70 ℃, and fully stirring for dissolving to obtain a mixture B;

when the temperature of the mixture B is reduced to 40 ℃, adding 0.1g of fibrous adhesion egg yolk, 0.02g of methyl hydroxybenzoate and 0.06g of propyl hydroxybenzoate while stirring to obtain a mixture C;

mixing and stirring the mixture A, C uniformly at 40 ℃, homogenizing for 5min at the rotation speed of 1500 rpm to obtain a mixture D;

and adding triethanolamine into the mixture D, stirring uniformly, filling the gel final product into a vaginal push tube, and filling 3.5g of gel into each tube to obtain the cold compress gel which can effectively treat stress urinary incontinence caused by pelvic floor muscle relaxation.

Example 3

Weighing 1.6g of curcuma zedoary extract, 2g of fructus psoraleae, 2g of dendrobium officinale hydrolysate, 0.6g of boswellic acid and 0.6g of soybean isoflavone, and uniformly dispersing the five raw materials by using 10g of propylene glycol to obtain a mixture A;

weighing 1g of carbomer U20 powder and 1.2g of hydrolyzed collagen, adding into 120g of deionized water, adding 10g of glycerol, heating to 80 ℃, and fully stirring for dissolving to obtain a mixture B;

when the temperature of the mixture B is reduced to 45 ℃, adding 0.15g of fibrous adhesion egg yolk, 0.3g of methyl hydroxybenzoate and 0.3g of propyl hydroxybenzoate while stirring to obtain a mixture C;

mixing and stirring the mixture A, C uniformly at 45 ℃, homogenizing for 5min at the rotation speed of 1500 revolutions per minute to obtain a mixture D;

and adding triethanolamine into the mixture D, stirring uniformly, filling the gel final product into a vaginal push tube, and filling 3.5g of gel into each tube to obtain the cold compress gel which can effectively treat stress urinary incontinence caused by pelvic floor muscle relaxation.

The use method of the cold compress gel prepared by the invention comprises the following steps: before use, the vulva part is cleaned, the hands are repeatedly cleaned, the gel packaging bag is torn open, the injector with the gel is taken out, the front cover at the front end of the injector is taken down and inserted into the injector to be used as a piston push rod; the injector with the gel is slowly inserted into the vagina by adopting a supine position and bending the knee, the piston push rod is pushed by the forefinger to push the gel into the vagina, the injector is drawn out after the injection is finished, and the injector is laid for 10 to 15 minutes to be fully absorbed.

And simultaneously tearing the second gel packaging bag, and applying the gel on Shenque acupoint, Guanyuan acupoint, Zusanli acupoint and Sanyinjiao acupoint to achieve better effect.

The use amount is as follows: vaginal bolus administration: one per day or one every other day; coating acupuncture points: one branch per day; effectively treat the stress urinary incontinence caused by the relaxation of pelvic floor muscles.

Test examples

There is no corresponding animal model for pelvic floor muscle relaxation:

(1) we used the effect of isolated rat colon smooth muscle contraction movement as the observation object and initially studied its mechanism of action.

The method comprises the following steps: rat colon smooth muscle strips were prepared, and the contraction effect of the cold compress gels prepared in example 1 at different concentrations on colon smooth muscle was observed with 10g/L NaCl Solution (NS) as a control group; the contraction effect of the cold compress gel on the muscle strips after the muscle strips are incubated at 1.0g, 3.5g and 7g of 3 doses by taking the cold compress gel as a control group is observed. The results are shown in Table 1:

TABLE 1 Effect of different doses of cold-applied gel on spontaneous contraction of rat colon smooth muscle

As a result: the cold compress gel large dose group has obvious promotion effect on isolated rat colon smooth muscle, and compared with NS control group, the cold compress gel 3.5gm and 7gm induced muscle maximum contraction amplitude and area under the curve have statistical significance. And (4) conclusion: the cold compress gel prepared in example 1 has an excitatory effect on the contractile activity of the rat colon smooth muscles and is positively correlated to the dose.

(2) The cold compress gel prepared in example 1 was used to study the effect of promoting muscle injury repair using C2C12 myoblasts and rat skeletal muscle in vitro experimental system.

The method comprises the following steps: C2C12 cells were treated with 3 different doses of cold compress gel, 1.0g, 3.5g, 7g, and scored and cell proteins were extracted to detect EGFR expression levels. Rat supratrochlear muscles were surgically removed, cultured in vitro with cold-compress gel and tested for EGFR expression levels.

Effect on C2C12 cell mobility: C2C12 cells were treated with 3 different doses of cold compress gel, 1.0g, 3.5g, 7g, for BCA protein quantification, chemiluminescence reagent development, and exposure in an exposure machine. All statistical data were analyzed for significance using SPSS software ANOVA.

FIG. 1 is a graph of scratch test conducted on different groups of treated C2C12 myoblasts; the results of the cell scratching experiments in fig. 1C2C12 show that the cell migration in the cold compress gel (3.5g) group is significantly increased (fig. 1.F), and the difference is statistically significant compared with the blank control group. The cell migration was slightly higher in the 1g and 7g cold-compressed gel groups than in the blank control group, but the difference was not significant.

FIG. 2 is a graph showing the expression level of EGFR in C2C12 cells after 2h of treatment in different groups, and FIG. 2 shows that there is a certain tendency of increasing EGFR expression in C2C12 cells after cold compress gel treatment prepared in example 1. The results of culturing EGFR expression level in muscle tissue in vitro in C2C12 show that the cold compress gel prepared in example 1 can significantly increase EGFR expression level in muscle tissue.

FIG. 3 shows the migration rate of C2C12 cells after 2.5h treatment of different groups, and FIG. 3 shows that the cold compress gel prepared in example 1 has certain promotion effect on migration and proliferation of C2C12 after 2.5h treatment, and the effect is related to concentration.

And (4) conclusion: the experimental result shows that the cold compress gel prepared in the example 1 plays a role in promoting the proliferation of skeletal muscle cells by positively regulating the EGFR expression level of C2C12 cells and skeletal muscle tissues, and can play a certain role in repairing the motor injury of the skeletal muscle.

(3) The invention adopts a castration method to establish a rat VA model, and 40 rats are divided into a pseudo-operation group and an experimental group (10 models, 10 collagen (cold compress gel) groups and 10 estrogens) by a random digital table method. The sham operation group: opening abdominal cavity, exposing in air for 30min, and closing abdomen; the collagen group is administered to vagina at 28 days after operation for 3.5g of cold compress gel once a day for 14d continuously; the estrogen group was administered once daily for 14 days, 0.5mL of estrogen per day vaginally, starting on day 28 after surgery. Experimental groups: the cold compress gel group prepared in example 1 and the estrogen only group were administered to the vagina continuously for 14d at 4 weeks after castration. Measuring the change of pH value of rats, the uterine mass and the vaginal epithelial thickness, and detecting the expression of TGF-beta 1, Smad2/3 and p-Smad2/3 proteins by Western blot. The Western blot detection method comprises the following steps: adding 1mL of RIPA and 1 μ L of PMSF lysis solution into each group of tissue, cutting tissue, homogenizing under tissue grinder, standing on ice for 5min, and centrifuging at 13000r/min for 15 min. Taking supernatant, then measuring the protein concentration by using a BCA kit, then, balancing and adding a total volume of 1/4 sample buffer solution, heating and boiling to inactivate the protein, carrying out electrophoresis and membrane transfer on 10% gel by using the protein, carrying out room-temperature sealing on skim milk powder, carrying out primary antibody 4T incubation overnight, washing the membrane for 3 times by using TBST, then carrying out secondary antibody incubation, and developing.

Table 2: vaginal pH of rats of different groups (n ═ 10, x- ± s)

a: p <0.05, compared to week 0

Table 3 uterine mass and vaginal epithelium thickness (n ═ 10, x ± 5) for each group of rats

a: p <0.05, compared to sham group; b: p <0.05, compared to estrogen group; c: p <0.001, compared to model group

As a result: according to the evaluation of the vaginal pH value, the pH values of the collagen group (cold compress gel) and the estrogen group after treatment have a descending trend (Table 2), the difference is statistically significant (P is less than 0.05) compared with the pH value before treatment, and the uterus quality measurement result of rats shows that the uterus quality of the model group and the collagen group is obviously reduced (P is less than 0.05) compared with the sham operation group; the uterine mass was significantly increased in the estrogen group compared to the model group, the collagen group (table 4, P < 0.05). After treatment with the cold compress gel prepared in example 1, the vaginal pH of castrated rats decreased; increased epithelial thickness, increased proliferative capacity; the synthesis of extracellular matrix components is increased, and the degradation is reduced; phosphorylated Smad2/3 protein expression is increased in vaginal tissue. The cold compress gel group prepared in example 1 has higher uterine safety compared to the group treated with estrogen alone.

And (4) conclusion: the cold compress gel prepared in example 1 can effectively relieve vaginal atrophy, increase extracellular matrix components in vaginal tissues and promote repair of vaginal epithelium.

(4) The actual use of the cold compress gel prepared in example 1 was confirmed in the present invention using pilot research (also known as "preliminary research"). Is a preliminary study for solving the problems of the method and procedure in the formal study. The entire procedure required for the full study, including the final data analysis, is performed. The total number of subjects was much less than that of the official study. The general effects are as follows: 1) the data collection method is improved, and control variables and dependent variables which need to be added are provided. The existing measurement techniques are partially modified. 2) The hypothesis is pre-checked to make the hypothesis more accurate and appropriate. If necessary, the original assumptions can be removed and new assumptions can be made. 3) Clues or insights are obtained that are useful in the point of view, method, etc. of the official experiment. 4) The planned statistical and analytical steps are examined to see if they are useful for solving the subject of the study. 5) And the manpower and material resources required to be spent according to the original plan are inspected, so that the formal experiment is more economical. A decision can even be made on the feasibility problem of the experimental plan. 6) Feedback is obtained from the subject regarding the indicators, strength of the independent variables, etc.

Pilot test:

the method comprises the following steps: 46 patients are selected, the average age is 47 years (38-65 years), the course of disease is 2-15 years, the average 7 years are all women who have passed through the woman, the number of times of delivery is 2-4, and the induction test and the bladder neck elevation test are positive. Before treatment, urodynamics and urine routine are checked, and urination abnormality caused by lower urinary tract obstruction and urinary system infection is eliminated.

Diagnostic criteria:

1. the SUI clinical diagnostic criteria refer to the Gullen scale. (1) I degree: when abdominal pressure suddenly increases, such as cough, urinary incontinence occasionally occurs; (2) II degree: urinary incontinence occurs with each cough, breath hold or exertion; (3) and (3) III degree: incontinence of urine occurs when the patient walks and stands; (4)1V degree: urinary incontinence also occurs when the patient is in bed.

2. During the treatment period, advising the patient to record a urination diary and recording the daily drinking water, urination and urine leakage conditions; questionnaires are filled in during the first and second visits. Performing a urine pad test, wherein the patient is ordered to drink 500ml of water 15min before the test, the patient is ordered to walk or climb stairs for 30min, stand up for 10 times, cough for 1O times, run for 1min, bend down for 5 times and wash hands for 1min during the test, the increased weight of the urine pad of the patient is weighed, and the urine leakage amount is more than or equal to 2g in 1h and is positive; mild to moderate: the urine leakage is less than 10g when the urine leakage is less than or equal to 1h after 2 g; and (3) severe degree: the urine leakage amount is less than 50g when the 10g is less than or equal to 1 h; extremely severe: the urine leakage is more than or equal to 50g in 1 h.

Treatment: vaginal bolus administration of the cold compress gel prepared in example 1: one treatment daily for 14 days followed by one treatment every other day for 14 days; it can be applied to SHENQUE acupoint, GUANYUAN acupoint, Zusanli acupoint, and SANYIJIAO acupoint once a day.

Evaluation:

(1) and (5) detecting the contraction condition of the pelvic floor muscles. The examiner lightly presses the deep muscles from the posterior vaginal wall to the pelvic floor with the index and middle fingers to evaluate the contraction thereof, thereby judging the strength of the muscles and the degree of fatigue. And carrying out manual measurement on the pelvic floor muscle strength by adopting Oxford scoring, wherein the detection result is divided into six grades of O-5, wherein the grades of 0, 1 and 2 are patients with pelvic floor muscle relaxation, and the grades of 3, 4 and 5 are normal patients. The method has the following advantages: the grade of the patient is raised by one grade after treatment; and (4) invalidation: patient grade did not change after treatment. The statistical method comprises the following steps: SPSS16.0 software is used for data analysis, the measurement data is expressed by x-s, the comparison before and after treatment is tested by x2, and the statistical significance is achieved when P is less than O.05.

(2) The urine leakage is more than or equal to 2g after 1 hour, and the urine is positive; mild to moderate: the urine leakage is less than 10g when the urine leakage is less than or equal to 1h after 2 g; and (3) severe degree: the urine leakage amount is less than 50g when the 10g is less than or equal to 1 h; extremely severe: the urine leakage is more than or equal to 50g in 1 h. And (3) curing: the urine leakage amount is less than or equal to 2g after 1 hour, and the effect is as follows: the urine leakage is reduced and is converted to the first level; and (4) invalidation: there was no change.

As a result:

(1) after a treatment course, the detection result of the comprehensive muscle strength of the pelvic floor muscles of 38 patients in 46 patients reaches more than 3 grades. The normal rate reaches 82.60%, 43 patients are effective, the effective rate is 93.47%, only 3 patients are not improved, and the difference between before and after treatment is obvious (P <0.05)

(2) Urine pad test results: 33 cases were cured (71.74%), 10 cases were improved (21.74%), 3 cases were not effective (6.52%); the total effective rate is 93.48% (43/46).

For discussion of this study:

stress incontinence is a common group of disorders in which the pelvic floor function of the body is impaired, with a high incidence in adult women. The occurrence of the diseases shows a rising trend along with the age of the patients, and the serious influence is brought to the physical and psychological health of the patients. The development of the disease is related to factors such as prolonged labor process, obstetric forceps assisted delivery, urgent labor and the like in the process of delivery, ligament, levator ani and other pelvic floor tissues are lost in the process of delivery, the pelvic floor tissues are loosened, the bladder neck moves downwards towards the direction close to the urethra, the urethra is shortened, and urine leakage occurs when cough or other pressure in the abdominal cavity is increased.

The clinical treatment is mainly to recover the functions of pelvic floor muscles and improve the stability of detrusor. The cold compress gel prepared by the invention is used for active treatment and guiding a patient to actively carry out Kegel exercise, and the contraction function of pelvic floor muscles is improved. Through pelvic floor rehabilitation, the utility model is helpful to relax and contract the muscles around the anus, improve the elasticity of the muscles at the part or far away from the anus, effectively improve the closing pressure of the urethra and improve stress urinary incontinence.

The embodiments in the present description are described in a progressive manner, each embodiment focuses on differences from other embodiments, and the same and similar parts among the embodiments are referred to each other. The device disclosed by the embodiment corresponds to the method disclosed by the embodiment, so that the description is simple, and the relevant points can be referred to the method part for description.

The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.