阅读说明：本技术 一种地高辛在防治运动病、梅尼埃病药物中的应用 (Application of digoxin in preventing and treating motion sickness and Meniere disease ) 是由 周鑫 姜正林 徐丽华 任畅 于 2021-10-18 设计创作，主要内容包括：本申请公开了一种地高辛在防治运动病、梅尼埃病药物中的应用,效果实验采用的实验动物为Sprague-Dawley(SD)大鼠,体重200-220 g,雌性,适应性饲养3天,然后进行旋转刺激诱导运动病和梅尼病两个晕模型的实验,以条件性味觉厌恶为模型；再进行地高辛的抗晕实验,以旋转刺激诱导的条件性味觉厌恶为模型,检测旋转刺激后两天大鼠对0.15%糖精钠溶液饮用量的变化；应用地高辛来拮抗旋转刺激和升高的精氨酸加压素、盐皮质激素醛固酮对内耳的作用,拮抗内耳内淋巴液生成过量,降低前庭刺激敏感性,抑制运动病的发生和抑制梅尼埃病程进展。(The application discloses application of digoxin in drugs for preventing and treating motion sickness and Meniere disease, wherein an effect experiment adopts Sprague-Dawley (SD) rats with the weight of 200-220g and female animals which are bred adaptively for 3 days, and then an experiment of inducing two motion sickness and Meniere disease dizziness models by rotary stimulation is carried out, and conditioned taste aversion is taken as a model; then carrying out digoxin anti-dizziness experiment, taking the conditioned taste aversion induced by the rotary stimulation as a model, and detecting the change of the drinking amount of the 0.15% saccharin sodium solution of the rats on two days after the rotary stimulation; the application of digoxin to antagonize the effects of rotation stimulation and increased arginine vasopressin and mineralocorticoid aldosterone on the inner ear, antagonize excessive production of lymph fluid in the inner ear, reduce the sensitivity of vestibular stimulation, inhibit the occurrence of motion sickness and inhibit the progress of Meniere's disease.)

1. The application of digoxin in preventing and treating motion sickness and Meniere disease is characterized in that: the application effect experiment of the digoxin in the medicines for preventing and treating the motion sickness and the Meniere disease adopts an experimental animal Sprague-Dawley (SD) rat with the weight of 200-220g and female for 3 days of adaptive breeding, and then the experiment of inducing two halo models of the motion sickness and the Meniere disease by rotary stimulation is carried out, and the conditioned taste aversion is used as the model; then carrying out digoxin anti-dizziness experiment, taking the conditioned taste aversion induced by the rotating stimulation as a model, and specifically comprising the following steps:

step 1: allowing the rats to drink freely in 0.15% saccharin sodium solution, measuring the drinking amount once every 24h, and administering digoxin by intraperitoneal injection 1h before rotary stimulation, wherein the dosage is 1-20 mg/kg;

step 2, rotational stimulation induces conditioned taste aversion in rats: the rotation stimulation induces the SD rat to generate the conditional taste aversion (halo), and the judgment is carried out by measuring the reduction of the intake of 0.15 percent Saccharin Sodium Solution (SSS) by the rat;

step 3, inhibition of rotation-stimulus induced conditioned taste aversion behaviors by digoxin: digoxin (10mg/kg) was administered by intraperitoneal injection 1h before the SD rats were subjected to 2h of rotational stimulation, and the change in the drinking amount of the 0.15% saccharin sodium solution by the rats two days after the rotational stimulation was examined compared with that before the rotational stimulation.

2. The use of digoxin as set forth in claim 1 for preventing and treating motion sickness and meniere's disease, characterized in that: the digoxin (amioride, Aml) was purchased from Sigma Aldrich, west coden, manufactured by tianjin north food limited, and the SD rats were provided by the university of south china laboratory animals center.

3. The use of digoxin as set forth in claim 1 or 2 for preventing and treating motion sickness and meniere's disease, characterized in that: in the step 2, the rat is freely and unrestrained placed in a rotary stimulation device embedded with an upper layer of partition board and a lower layer of partition board, each partition board is evenly divided into 6 parts, 1 partition board is placed in each part, the device slowly rotates clockwise around a vertical shaft at a constant speed, rotates around a horizontal shaft at a variable speed, and firstly rotates at an angular acceleration of 16 degrees/s²Accelerating until the angular velocity reaches the maximum value of 120 DEG/s, and immediately controlling the angular velocity to be-48 DEG/s²The angular acceleration of the device is decelerated until the device stops, 10s is a period, then the period is repeated anticlockwise, the stimulation is repeated for 120min, the drinking amount of each SD rat to the 0.15% saccharin sodium solution in two days before and two days after the rotation stimulation is continuously monitored, corresponding experimental data are recorded, the intake change of the saccharin sodium solution is calculated, and the index is as follows: the change percentage of the drinking amount of the saccharin sodium solution is (the average drinking amount of the saccharin sodium solution after 2 days of rotation-2 balance average drinking amounts before rotation)/2 balance average drinking amounts before rotation, then statistical analysis is carried out, and independent sample t test (Student's test) is adopted for comparison between the two groups, and more than two groups are addedThe comparison between groups is carried out pairwise by adopting One-way analysis of variance (One way anova) and an LSD method, and when p is<At 0.05, the difference was considered significant, i.e., statistically significant.

4. The use of digoxin as set forth in claim 3 for preventing and treating motion sickness and Meniere's disease, characterized in that: the rat is freely and unrestrained in the rotary stimulation device embedded with an upper layer of partition board and a lower layer of partition board according to the report method of Crampton and Lucot (1985), each partition board is averagely divided into 6 parts, and 1 rat is placed in each part.

Technical Field

The invention belongs to the technical field of biological medicines, and particularly relates to application of digoxin in medicines for preventing and treating motion sickness and Meniere's disease.

Background

When a person moves passively, the person is stimulated by abnormal vestibule stimulation or/and abnormal vision stimulation, and can have motion sickness which is mainly manifested by pale complexion, cold sweat, salivation, indifference, abdominal discomfort, dizziness, nausea, vomiting and the like. At present, the detailed pathogenesis of the motion sickness is still not clear, and the prevention and treatment aspect still lacks of drugs with high efficiency and no side effect. The most commonly used medicine for preventing and treating motion sickness at present is antihistaminic medicine dimenhydrinate (dimenhydrinate), and the single medicine with the best effect is anticholinergic medicine scopolamine, but the effective action parts of the medicines are not very clear, the side effects are also large, especially the side effects of central inhibition, blurred vision, memory impairment and the like influence the operation of people under various special environmental conditions such as navigation, aerospace and the like.

Meniere's disease is a sudden onset of rotational vertigo accompanied by symptoms such as tinnitus, deafness, fullness in the head or ear, and the like, and severe vertigo may be accompanied by symptoms such as nausea, vomiting, cold sweating, and the like. The basic pathology of the disease is altered to membranous labyrinth hydroncus. The research finds that the meniere disease is related to factors such as immune response, vegetative nerve functional disturbance or infection, and the like, and the factors interfere the function of capillary vessels and veins to cause obstruction of backflow of capillaries and veins, or cause obstruction of lymphatic vessels and malabsorption of lymph fluid in the inner ear to cause membrane labyrinth hydrops and cause imbalance of balance function of the inner ear. Meniere's disease is usually acute and repeated, and is a difficult treatment in the ear, nose and throat department. For the treatment of meniere's disease, the etiology has not been completely clarified so far, so at present, clinically, the pathophysiology process is mainly based on symptomatic treatment, and therapeutic measures such as anti-dizziness, sedation, blood vessel dilation, dehydration and supporting therapy are adopted, or destructive surgery or non-surgical treatment is adopted, and when conservative treatment is ineffective, surgical treatment is considered, such as endolymphatic sac decompression or shunt surgery, vestibular nerve amputation, labyrinthine damage or excision and the like. These methods either affect the therapeutic effect or bring serious side effects such as unnecessary or even impaired hearing and balance function.

Many studies find that the motion sickness and Meniere's disease cause excessive lymph formation in the inner ear, and the reabsorption of water by the epithelium of the inner lymph sac is reduced, which leads to water accumulation in the membranous labyrinth, and finally induces symptoms such as dizziness, nausea, vomiting, and the like. Therefore, if a drug is used to inhibit the formation of endolymphatic excess in the inner ear, it can be applied to the development of new drugs for motion sickness and meniere's disease.

Aldosterone is an important hormone involved in water-salt balance and maintaining the homeostasis of the body. Research shows that the phenomenon of membrane labyrinth hydrops of cochlea of guinea pigs is caused by injecting aldosterone into abdominal cavity, and the suggestion that aldosterone probably promotes lymphangiogenesis in inner ear and/or reduces endolymph reabsorption, thereby having the effect of increasing endolymph volume. In addition, it has been found that the membrane labyrinthine hydrops phenomenon induced by intraperitoneal aldosterone injection in cochlea of guinea pig causes epithelial sodium channel (alpha-ENaC) protein and sodium potassium pump (Na) in cochlear tissue⁺,K⁺-ATPase) is increased. These studies suggest that the mineralocorticoid aldosterone-regulated epithelial sodium channel and sodium potassium pump may influence the inner ear water-salt balance through the action of inner ear target proteins, thereby participating in the development of motion sickness and meniere's disease.

The sodium potassium pump can decompose ATP to obtain energy for a special protein in cell membrane, and Na is carried out by using the energy⁺、K⁺Active transport of (i.e. reverse concentration gradient of Na)⁺Transport of K from inside to outside of the cell⁺Transported into the cell from outside the cell, the primary function of ATPase is to control K inside and outside the cell membrane⁺，Na⁺The concentration difference of ions plays an extremely important role in maintaining the ion concentration difference between the inside and outside of the cell and the normal membrane potential. Sodium potassium pump inner and outer lymph liquid K⁺，Na⁺The concentration difference of the ions is important, and excessive Na in the cells is generated⁺The reverse chemical gradient is pumped out of the cell. When motion sickness and meniere's disease, sodium potassium pump expression and activation increase, eventually causing increased endolymphatics, resulting in membrane labyrinth hydrosis. Therefore, the medicine for inhibiting the transitional activation and expression of the sodium-potassium pump can be applied to motion sickness and Meniere disease.

The cardiac glycoside drugs are mainly used for clinically treating heart failure and some arrhythmia, the most representative of the cardiac glycoside drugs is Digoxin (Dig.), and the cardiac glycoside drugs can slightly inhibit a sodium-potassium pump under the condition of treatment dosage, destroy the active transport of sodium on a cell membrane and enable intracellular Na⁺Increase in concentration, K⁺The concentration is reduced. Research shows that digoxin can relieve edema of pancreas and lung of mice, so that digoxin which is an inhibitor of a sodium-potassium pump can be used for inhibiting the generation of lymph fluid in inner ear, relieving endolymphatic edema and relieving vestibule stimulation, and therefore a protein sodium-potassium pump at the downstream of a mineralocorticoid signal pathway is taken as a target point, and a novel anti-motion disease drug is developed from digoxin which antagonizes the sodium-potassium pump.

Disclosure of Invention

The technical problem to be solved is as follows:

aiming at the defects of the prior art, the application solves the problems that the effective action part of the existing medicine is not clear, the side effect is large, especially the side effects such as central inhibition, blurred vision, memory impairment and the like exist, and the medicine with high efficiency and no side effect is lacked; an application of digoxin in preventing and treating sports diseases and Meniere's disease is provided.

The technical scheme is as follows:

in order to achieve the purpose, the application is realized by the following technical scheme:

the adopted experimental animals are Sprague-Dawley (SD) rats with the weight of 200-220g and female animals, are bred adaptively for 3 days, and then are subjected to an experiment of inducing two halo models of the motion disease and the Meniere disease by rotating stimulation, and the conditioned taste aversion is taken as the model; then carrying out digoxin anti-dizziness experiment, taking the conditioned taste aversion induced by the rotating stimulation as a model, and comprising the following steps:

step 1: allowing the rats to drink freely in 0.15% saccharin sodium solution, measuring the drinking amount once every 24h, and administering digoxin by intraperitoneal injection 1h before rotary stimulation, wherein the dosage is 1-20 mg/kg;

step 2, rotational stimulation induces conditioned taste aversion in rats: the rotation stimulation induces the SD rat to generate the conditional taste aversion (halo), and the judgment is carried out by measuring the reduction of the intake of 0.15 percent Saccharin Sodium Solution (SSS) by the rat;

step 3, inhibition of rotation-stimulus induced conditioned taste aversion behaviors by digoxin: digoxin (10mg/kg) was administered by intraperitoneal injection 1h before the SD rats were subjected to 2h of rotational stimulation, and the change in the drinking amount of the 0.15% saccharin sodium solution by the rats two days after the rotational stimulation was examined compared with that before the rotational stimulation.

As a preferred technical scheme of the invention: the digoxin (amioride, Aml) was purchased from Sigma Aldrich, west coden, manufactured by tianjin north food limited, and the SD rats were provided by the university of south china laboratory animals center.

As a preferred technical scheme of the invention: in the step 2, the rat is freely and unrestrained placed in a rotary stimulation device embedded with an upper layer of partition board and a lower layer of partition board, each partition board is evenly divided into 6 parts, 1 partition board is placed in each part, the device slowly rotates clockwise around a vertical shaft at a constant speed, rotates around a horizontal shaft at a variable speed, and firstly rotates at an angular acceleration of 16 degrees/s²Accelerating until the angular velocity reaches the maximum value of 120 DEG/s, and immediately controlling the angular velocity to be-48 DEG/s²The angular acceleration of the device is decelerated until the device stops, 10s is a period, then the period is repeated anticlockwise, the stimulation is repeated for 120min, the drinking amount of each SD rat to the 0.15% saccharin sodium solution in two days before and two days after the rotation stimulation is continuously monitored, corresponding experimental data are recorded, the intake change of the saccharin sodium solution is calculated, and the index is as follows: changing the drinking amount of the saccharin sodium solution into percentage (average drinking amount of the saccharin sodium solution after 2 days of rotation-2 balance average drinking amounts before rotation)/2 balance average drinking amounts before rotation, then performing statistical analysis, adopting independent sample t test (Student's test) for comparison between two groups, adopting One-factor way ANOVA (One way ANOVA) for comparison between multiple groups, and performing pairwise comparison by an LSD method, when p is the average drinking amount of the saccharin sodium solution after rotation, performing pairwise comparison<At 0.05, the difference was considered significant, i.e., statistically significant.

As a preferred technical scheme of the invention: the rat is freely and unrestrained in the rotary stimulation device embedded with an upper layer of partition board and a lower layer of partition board according to the report method of Crampton and Lucot (1985), each partition board is averagely divided into 6 parts, and 1 rat is placed in each part.

Has the advantages that:

the application provides an application of digoxin in drugs for preventing and treating motion sickness and Meniere disease, and compared with the prior art, the digoxin has the following beneficial effects:

1. as an antagonist of aldosterone downstream target protein sodium potassium pump, digoxin intraperitoneal injection can relieve conditioned taste aversion caused by rotational stimulation, i.e. antagonize motion sickness and meniere's disease (dizziness);

2. since the pathogenesis of meniere's disease is directly related to the action of the endolymphatic fluid of the inner ear and aldosterone on the inner ear, the present study results also support the use of digoxin to treat meniere's disease.

3. The application of digoxin to antagonize the effects of rotation stimulation and increased arginine vasopressin and mineralocorticoid aldosterone on the inner ear, antagonize excessive production of lymph fluid in the inner ear, reduce the sensitivity of vestibular stimulation, inhibit the occurrence of motion sickness and inhibit the progress of Meniere's disease.

4. The research shows that the digoxin intraperitoneal injection can relieve the conditioned taste aversion caused by the rotational stimulation, namely antagonizing the motion sickness and the Meniere disease (areola).

Description of the drawings:

FIG. 1 is a graph showing the variation of the amount of the sodium saccharin solution in rats after intraperitoneal injection of digoxin in the present application.

Detailed Description

The following will further explain the embodiments and working procedures of the present invention by referring to examples.

Example 1:

the application of digoxin in preventing and treating motion sickness and Meniere disease is realized by adopting Sprague-Dawley (SD) rats with the weight of 200-; an antihalation experiment was performed with digoxin (amioride, Aml) purchased from Sigma Aldrich (Sigma-Aldrich) of west codex america, manufactured by tianjin north food ltd, and SD rats provided by the university of south china laboratory animals center, in a model of conditioned taste aversion induced by rotational stimulation, by the steps of:

step 1: allowing the rats to drink freely in 0.15% saccharin sodium solution, measuring the drinking amount once every 24h, and administering digoxin by intraperitoneal injection 1h before rotary stimulation, wherein the dosage is 10 mg/kg;

step 2, freely and unrestrained rats are placed into a rotary stimulation device embedded with an upper layer of partition plate and a lower layer of partition plate according to a report method of Crampton and Lucot (1985), each partition plate is evenly divided into 6 parts, 1 part of the partition plate is placed, the device slowly rotates clockwise around a vertical shaft at a constant speed and rotates around a horizontal shaft at a variable speed, and the device firstly rotates at an angular acceleration of 16 degrees/s²Accelerating until the angular velocity reaches the maximum value of 120 DEG/s, and immediately controlling the angular velocity to be-48 DEG/s²The angular acceleration of the device is decelerated until the device stops, 10s is a period, then the period is repeated anticlockwise, the stimulation is repeated for 120min, the drinking amount of each SD rat to the 0.15% saccharin sodium solution in two days before and two days after the rotation stimulation is continuously monitored, corresponding experimental data are recorded, the intake change of the saccharin sodium solution is calculated, and the index is as follows: changing the drinking amount of the saccharin sodium solution into percentage (average drinking amount of the saccharin sodium solution after 2 days of rotation-2 balance average drinking amounts before rotation)/2 balance average drinking amounts before rotation, then performing statistical analysis, adopting independent sample t test (Student's test) for comparison between two groups, adopting One-factor way ANOVA (One way ANOVA) for comparison between multiple groups, and performing pairwise comparison by an LSD method, when p is the average drinking amount of the saccharin sodium solution after rotation, performing pairwise comparison<At 0.05, the difference is considered significant, i.e. statistically significant;

step 3, inhibition of rotation-stimulus induced conditioned taste aversion behaviors by digoxin: digoxin (10mg/kg) was administered by intraperitoneal injection 1h before the SD rats were subjected to 2h of rotational stimulation, and the change in the drinking amount of the 0.15% saccharin sodium solution by the rats two days after the rotational stimulation was examined compared with that before the rotational stimulation.

As shown in fig. 1, the consumption of saccharin sodium solution was significantly reduced (p <0.05) in the Rotation-stimulated group (Rotation) SD rats compared to the Control group (Control), while the reduction of saccharin sodium solution in the Digoxin group (Digoxin + Rotation) compared to the Rotation-stimulated solvent group (Vehicle + Rotation) was suppressed (p < 0.05). The results show that the aldosterone downstream target protein sodium-potassium pump antagonist digoxin can inhibit the expression of conditioned taste aversion behaviors caused by rotation stimulation, namely, the motion disease and meniere disease (dizziness), and the digoxin can be used as a medicine for preventing and treating the motion disease and the meniere disease. The digoxin concentration is 10 mg/kg.

Example 2:

the application of digoxin in preventing and treating motion sickness and Meniere disease is realized by adopting Sprague-Dawley (SD) rats with the weight of 200-; an antihalation experiment was performed with digoxin (amioride, Aml) purchased from Sigma Aldrich (Sigma-Aldrich) of west codex america, manufactured by tianjin north food ltd, and SD rats provided by the university of south china laboratory animals center, in a model of conditioned taste aversion induced by rotational stimulation, by the steps of:

step 1: allowing the rats to drink freely in 0.15% saccharin sodium solution, measuring the drinking amount once every 24h, and administering digoxin by intraperitoneal injection 1h before rotary stimulation, wherein the dosage is 1 mg/kg;

step 2, freely and unrestrained rats are placed into a rotary stimulation device embedded with an upper layer of partition plate and a lower layer of partition plate according to a report method of Crampton and Lucot (1985), each partition plate is evenly divided into 6 parts, 1 part of the partition plate is placed, the device slowly rotates clockwise around a vertical shaft at a constant speed and rotates around a horizontal shaft at a variable speed, and the device firstly rotates at an angular acceleration of 16 degrees/s²Accelerating until the angular velocity reaches the maximum value of 120 DEG/s, and immediately controlling the angular velocity to be-48 DEG/s²The angular acceleration of the device is decelerated until the device stops, 10s is a period, then the period is repeated anticlockwise, the stimulation is repeated for 120min, the drinking amount of each SD rat to the 0.15% saccharin sodium solution in two days before and two days after the rotation stimulation is continuously monitored, corresponding experimental data are recorded, the intake change of the saccharin sodium solution is calculated, and the index is as follows: change percentage of the saccharin sodium solution drinking amount (average drinking amount of saccharin sodium solution 2 days after rotation-2 balance drinking amounts before rotation)/2 before rotationBalance average drinking amount, performing statistical analysis, comparing two groups with independent sample t test (Student's test), comparing multiple groups with One-way ANOVA (One way ANOVA), and comparing two groups with LSD method, when p is<At 0.05, the difference is considered significant, i.e. statistically significant;

step 3, inhibition of rotation-stimulus induced conditioned taste aversion behaviors by digoxin: digoxin (10mg/kg) was administered by intraperitoneal injection 1h before the SD rats were subjected to 2h of rotational stimulation, and the change in the drinking amount of the 0.15% saccharin sodium solution by the rats two days after the rotational stimulation was examined compared with that before the rotational stimulation.

Example 3:

the application of digoxin in preventing and treating motion sickness and Meniere disease is realized by adopting Sprague-Dawley (SD) rats with the weight of 200-; an antihalation experiment was performed with digoxin (amioride, Aml) purchased from Sigma Aldrich (Sigma-Aldrich) of west codex america, manufactured by tianjin north food ltd, and SD rats provided by the university of south china laboratory animals center, in a model of conditioned taste aversion induced by rotational stimulation, by the steps of:

step 1: allowing the rats to drink freely in 0.15% saccharin sodium solution, measuring the drinking amount once every 24h, and administering digoxin by intraperitoneal injection 1h before rotary stimulation, wherein the dosage is 20 mg/kg;

step 2, freely and unrestrained rats are placed into a rotary stimulation device embedded with an upper layer of partition plate and a lower layer of partition plate according to a report method of Crampton and Lucot (1985), each partition plate is evenly divided into 6 parts, 1 part of the partition plate is placed, the device slowly rotates clockwise around a vertical shaft at a constant speed and rotates around a horizontal shaft at a variable speed, and the device firstly rotates at an angular acceleration of 16 degrees/s²Accelerating until the angular velocity reaches the maximum value of 120 DEG/s, and immediately controlling the angular velocity to be-48 DEG/s²Until stopped, for 10s for a period, and then repeating the period in the counterclockwise direction, thus repeatedly stimulating for 120min, and continuously monitoringThe drinking amount of each SD rat to the 0.15% saccharin sodium solution in two days before and two days after the rotary stimulation is recorded, corresponding experimental data are recorded, and the intake change of the saccharin sodium solution is calculated, wherein the indexes are as follows: changing the drinking amount of the saccharin sodium solution into percentage (average drinking amount of the saccharin sodium solution after 2 days of rotation-2 balance average drinking amounts before rotation)/2 balance average drinking amounts before rotation, then performing statistical analysis, adopting independent sample t test (Student's test) for comparison between two groups, adopting One-factor way ANOVA (One way ANOVA) for comparison between multiple groups, and performing pairwise comparison by an LSD method, when p is the average drinking amount of the saccharin sodium solution after rotation, performing pairwise comparison<At 0.05, the difference is considered significant, i.e. statistically significant;

step 3, inhibition of rotation-stimulus induced conditioned taste aversion behaviors by digoxin: digoxin (10mg/kg) was administered by intraperitoneal injection 1h before the SD rats were subjected to 2h of rotational stimulation, and the change in the drinking amount of the 0.15% saccharin sodium solution by the rats two days after the rotational stimulation was examined compared with that before the rotational stimulation.

Finally, the above embodiments are only for illustrating the technical solutions of the present invention and not for limiting, and other modifications or equivalent substitutions made by the technical solutions of the present invention by the person skilled in the art shall be covered by the scope of the claims of the present invention without departing from the spirit and scope of the technical solutions of the present invention.