阅读说明：本技术 一种沙蓬提取物和提取方法及其应用 (Salicornia bigelovii extract, and extraction method and application thereof ) 是由 包书茵 苏维恒 奥·乌力吉 王秀枝 于 2020-08-21 设计创作，主要内容包括：本发明公开了一种沙蓬提取物和提取方法及其应用,其特征在于沙蓬提取物用于对心脏损伤的保护,特别是异丙肾上腺素诱导的心肌损伤的保护,本发明的有益效果是：沙蓬低聚糖能够通过维持心肌细胞完整性,减轻心肌缺血,改善异丙肾上腺素诱导的急性心肌损伤,沙蓬低聚糖能够对于异丙肾上腺素多种病理机制诱导心肌损伤均具有良好的效果,沙蓬低聚糖对异丙肾上腺素多种病理机制所致心肌损伤均具有一定的改善作用,对心肌损伤具有一定的保护作用,是一种新型、高效、低毒的,针对异丙肾上腺素诱导的急性心肌损伤的多靶点调节作用防治药物。(The invention discloses an agriophyllum squarrosum extract, an extraction method and application thereof, which are characterized in that the agriophyllum squarrosum extract is used for protecting heart injury, in particular to protecting cardiac muscle injury induced by isoproterenol, and the invention has the beneficial effects that: the suaeda salsa oligosaccharide can be used for relieving myocardial ischemia and improving acute myocardial injury induced by isoproterenol by maintaining the integrity of myocardial cells, has a good effect on the myocardial injury induced by various pathological mechanisms of isoproterenol, has a certain improvement effect on the myocardial injury caused by various pathological mechanisms of isoproterenol, has a certain protection effect on the myocardial injury, is a novel, high-efficiency and low-toxicity multi-target regulation effect prevention and treatment medicine for the acute myocardial injury induced by isoproterenol.)

1. An agriophyllum squarrosum extract is characterized in that the main component of the agriophyllum squarrosum extract is agriophyllum squarrosum oligosaccharide.

2. An extraction method of an agriophyllum squarrosum extract is characterized in that the agriophyllum squarrosum extract is prepared according to the claim 1, the extraction method comprises the steps of crushing the agriophyllum squarrosum, adding ethanol for reflux extraction, filtering an extracting solution, combining filtrate, decompressing, recovering the ethanol, adding equal volume of water for dilution, filtering, passing the filtrate through an adsorption resin column, eluting with water, collecting water eluent, decompressing and concentrating to obtain the agriophyllum squarrosum extract.

3. The method for extracting Salicornia bigelovii Torr extract as claimed in claim 2, comprising: pulverizing herba Salicornia Herbacea into coarse powder, sequentially adding 10, 8 times of 95% ethanol, reflux-extracting for 2 times, each for 2 hr, mixing extractive solutions, removing residue, defatting medicinal powder, sequentially adding 10, 8 times of 70% ethanol, reflux-extracting for 3 times, 2 hr for the first time, 1 hr for the second time and 1 hr for the third time, filtering extractive solutions, mixing filtrates, recovering ethanol under reduced pressure, concentrating to no alcohol smell (1g:1ml), adding equal volume of water, diluting, and standing at low temperature overnight; filtering, passing the filtrate through D101 type macroporous adsorbent resin column, eluting with appropriate amount of water, collecting water eluate, concentrating under reduced pressure, decolorizing with active carbon, refining, concentrating under reduced pressure to obtain extract, drying under reduced pressure, and pulverizing to obtain herba Salicornia Bicoloris extract.

4. Use of an extract of Salicornia bigelovii as claimed in claim 2 or 3 for protecting against heart damage.

5. The use of an Salicornia bigelovii extract as claimed in claim 4, wherein said cardiac injury is isoproterenol-induced myocardial injury.

The technical field is as follows:

the invention belongs to the technical field of natural medicines, and particularly relates to a samphire extract, an extraction method and application thereof.

Background art:

cardiovascular diseases are the most common diseases in human beings, and have become the first leading cause of death in the world population today. Myocardial damage is one of the cardiovascular system diseases, and the proportion of the cardiovascular system diseases is higher and higher. Myocardial damage is the damage of myocardial cells, i.e. myocardial cell inflammation, necrosis, degenerative edema.

The cause of myocardial damage may be caused by coronary artery disease caused by severe ischemia, such as acute coronary syndrome, coronary heart disease, and acute myocardial infarction. Or myocardial damage caused by inflammation, such as myocarditis and pericarditis.

Yet another category is the result of myocardial stress, both emotional and physical, including, for example, the loss of a loved one, divorce or some property problem, such as the feeling of mental fragmentation, which actually causes myocardial damage;

the stress of the body, such as car accidents or major surgeries, is the result of myocardial stress, mainly caused by the increase of the level of hormones in the body, which can cause hypertension and myocardial infarction, resulting in myocardial damage;

still other autoimmune diseases involving the heart, such as lupus heart disease secondary to systemic lupus erythematosus, can also cause myocardial damage.

Such as physical or chemical damages, such as knife-gun stabbing of the heart muscle, or drugs that have toxic effects on the heart, such as drugs that treat tumors, drugs that treat hematological disorders, and drugs that also have toxic effects on the heart, causing myocardial damage.

There may be no obvious symptoms when the condition of myocardial cell injury is mild, but in those patients with severe condition, the patients may have the symptoms of chest distress, chest pain, dyspnea, asthenia, nausea, emesis, acute heart failure, even malignant arrhythmia, pericardial rupture and even sudden death.

Therefore, once myocardial damage is caused, the severity of the disease must be determined, and the degree of myocardial damage can be evaluated by an electrocardiogram and some special myocardial damage markers.

Therefore, since there may be no obvious symptoms when the condition of the damaged myocardial cells is mild, and the reasons for the myocardial damage are many and complicated, the prevention and treatment of the myocardial damage is always a big problem of cardiovascular system diseases, especially isoproterenol induced myocardial damage, which can induce the above mentioned various types of myocardial damage.

Isoproterenol is a strong beta receptor stimulant and can increase myocardial oxygen consumption through links of accelerating heart rate, enhancing myocardial contractility and the like, and causes cardiac overload, myocardial microcirculation disturbance, coronary artery spasm, myocardial infarction-like change, myocardial necrosis and even sudden death. These pathological lesions may be associated with disorders of mitochondrial energy metabolism, activation of apoptotic genes, oxidative stress, and the like.

The isoproterenol-induced myocardial injury has various pathological manifestations and complex pathological mechanism, so that no good specific medicine can be used for treating isoproterenol-induced myocardial injury. The search for novel, efficient and low-toxicity preventive and therapeutic drugs with multi-target regulation functions is still one of the key points of pharmaceutical research.

The invention content is as follows:

in order to solve the problems and overcome the defects of the prior art, the invention provides the salicornia mongolica extract, the extraction method and the application thereof, and can effectively solve the problem that no good specific medicine is available for myocardial injury induced by various pathologies of isoproterenol.

The specific technical scheme for solving the technical problems comprises the following steps: the agriophyllum squarrosum extract is characterized in that the main component of the agriophyllum squarrosum extract is agriophyllum squarrosum oligosaccharide.

The extraction method of the samphire extract is characterized in that the samphire extract is prepared by the extraction method, and the extraction method comprises the steps of smashing samphire, adding ethanol for reflux extraction, filtering an extracting solution, combining filtrate, recovering ethanol under reduced pressure, adding equal volume of water for dilution, filtering, passing the filtrate through an adsorption resin column, eluting with water, collecting water eluent, and concentrating under reduced pressure to obtain the samphire extract.

The extraction method of the agriophyllum squarrosum extract is characterized by comprising the steps of crushing the agriophyllum squarrosum into coarse powder, sequentially adding 10-8 times of 95% ethanol for reflux extraction for 2 times, 2 hours each time, combining extracting solutions, removing residues, degreasing medicinal powder, sequentially adding 10-8 times of 70% ethanol for reflux extraction for 3 times, 2 hours for the first time, 1 hour for the second time and 1 hour for the third time, filtering the extracting solutions, combining filtrates, decompressing, recovering ethanol, concentrating until no alcohol smell exists (1g:1ml), adding water with equal volume for dilution, and standing overnight at low temperature; filtering, passing the filtrate through D101 type macroporous adsorbent resin column, eluting with appropriate amount of water, collecting water eluate, concentrating under reduced pressure, decolorizing with active carbon, refining, concentrating under reduced pressure to obtain extract, drying under reduced pressure, and pulverizing to obtain herba Salicornia Bicoloris extract.

The application of the agriophyllum squarrosum extract is characterized in that the agriophyllum squarrosum extract is prepared by the extraction method of the agriophyllum squarrosum extract and is used for protecting heart injury.

The application of the Salicornia bigelovii extract is characterized in that the heart injury is isoproterenol-induced myocardial injury.

The application of the agriophyllum squarrosum extract is characterized in that the agriophyllum squarrosum extract is used for protecting various pathological myocardial injuries induced by isoproterenol, wherein the myocardial injuries comprise myocardial cell necrosis myocardial injury, myocardial ischemia myocardial injury and tachycardia myocardial injury, and the myocardial injury of the level of isoproterenol-induced hormone can be improved.

The invention has the beneficial effects that:

the invention obtains a natural substance of the salicornia mongolica oligosaccharide which can be used for protecting the isoproterenol-induced myocardial damage through an extraction process;

the agriophyllum squarrosum oligosaccharide can relieve myocardial ischemia and improve the acute myocardial injury induced by isoproterenol by maintaining the integrity of myocardial cells, has good effect on the myocardial injury induced by various pathological mechanisms of isoproterenol,

the agriophyllum squarrosum oligosaccharide has certain improvement effect on myocardial injury caused by various pathological mechanisms of isoproterenol, has certain protection effect on the myocardial injury, and is a novel, high-efficiency and low-toxicity medicine for preventing and treating acute myocardial injury induced by isoproterenol with multi-target regulation effect.

Description of the drawings:

FIG. 1 is an electrocardiogram of the normal control group of rats before administration;

FIG. 2 is an electrocardiogram of the normal control group of the present invention after the normal control group is applied with physiological saline;

FIG. 3 is an electrocardiogram of the rat after isoproterenol molding;

FIG. 4 is a low-dose electrocardiogram change of rat with Salicornia bigelovii oligosaccharide of the invention;

FIG. 5 is a electrocardiogram of the change of the electrocardiogram of rats in the amount of the suaeda salsa oligosaccharide;

FIG. 6 is a high-dose sarganin oligosaccharide electrocardiogram change electrocardiogram of a rat;

FIG. 7 is a drawing of a 200-fold HE stained histological section of a rat heart of the present invention; in the drawings:

the specific implementation mode is as follows:

in the description of the invention, specific details are given only to enable a full understanding of the embodiments of the invention, but it should be understood by those skilled in the art that the invention is not limited to these details for the implementation. In other instances, well-known structures and functions have not been described or shown in detail to avoid obscuring the points of the embodiments of the invention. The specific meanings of the above terms in the present invention can be understood in specific cases to those skilled in the art.

The specific implementation mode of the invention is as follows:

the main process conditions of the samphire extract are as follows: pulverizing herba Salicornia Herbacea, extracting with ethanol under reflux, filtering the extractive solution, mixing filtrates, recovering ethanol under reduced pressure, diluting with equal volume of water, filtering, passing the filtrate through adsorbent resin column, eluting with water, collecting water eluate, and concentrating under reduced pressure.

The process can be further optimized: pulverizing herba Salicornia Herbacea into coarse powder, sequentially extracting with 10 and 8 times of 95% ethanol under reflux for 2 times (each for 2 hr), mixing extractive solutions, and removing residue; sequentially adding 10, 8 and 8 times of 70% ethanol into defatted medicinal powder, reflux-extracting for 3 times, the first time for 2 hours, the second time for 1 hour and the third time for 1 hour, filtering the extractive solution, mixing the filtrates, recovering ethanol under reduced pressure, concentrating until no alcohol smell (1g:1ml), adding equal volume of water for dilution, and standing at low temperature overnight; filtering, passing the filtrate through D101 type macroporous adsorbent resin column, eluting with appropriate amount of water, and desorbing with 70% ethanol. Collecting water eluate, concentrating under reduced pressure, decolorizing with active carbon, refining, concentrating under reduced pressure to obtain extract, drying under reduced pressure, and pulverizing to obtain herba Salicornia Bicoloris extract.

In order to more intuitively show the process advantages of the invention, the process of the invention is briefly described as follows:

optimization of process conditions

1. Optimizing the ethanol extraction process conditions:

the samphire extract is extracted by an ethanol hot reflux method, and the ethanol concentration, the ethanol dosage, the extraction times and the extraction time are main factors influencing the extraction effect. Therefore, we chose L for each of the above factors at 3 levels₉(3⁴) Orthogonal table design test, using oligosaccharide extraction amount (total sugar) as evaluation index, researching and screening optimum process conditions:

first experiment design

By using L₉(3⁴) Orthogonal tables were experimentally designed and shown in tables 1 and 2 below.

TABLE 1 level table of factors for orthogonal test of ethanol reflux extraction process

Preparation of test solution

According to the orthogonal experimental design requirements of tables 1 and 3, precisely weighing 25g of sardine coarse powder, placing the sardine coarse powder in a 500ml round-bottom flask, adding B times of ethanol with A concentration, heating and extracting for C times by an electric heating sleeve, refluxing for D hours each time, performing suction filtration, combining filtrates, placing the combined filtrates in a 1000ml measuring flask, adding ethanol with A concentration to the scale, and shaking up to obtain the sardine coarse powder (each 1ml is equivalent to 0.025g of a raw material).

Determination of the content of the triose

The oligosaccharide, i.e. the total sugar content, is determined by phenol-sulfuric acid spectrophotometry.

Preparing reference solution by precisely weighing 25mg of anhydrous glucose reference substance dried at 105 ℃ to constant weight, precisely weighing, placing in a 100ml measuring flask, adding water to dissolve and dilute to scale, shaking up, accurately sucking 5ml, placing in a 10ml measuring flask, adding water to dilute to scale, and shaking up to obtain the final product.

Preparation of standard curve control solutions 0.1ml, 0.2ml, 0.3ml, 0.4ml, 0.5ml, 0.6ml were measured accurately, placed in 5ml test tubes with plug scale, added with water to 1.0ml each, and shaken well. Respectively adding 0.5ml of 5% phenol solution, mixing uniformly, quickly adding 3.5ml of sulfuric acid, shaking uniformly, keeping the temperature in a water bath at 40 ℃ for 30 minutes, taking out, cooling in an ice water bath for 5 minutes, taking out, taking a corresponding reagent as a blank, performing ultraviolet-visible spectrophotometry, measuring absorbance at 490nm wavelength, taking the absorbance as an ordinate, taking the concentration as an abscissa, drawing a standard curve, and obtaining the result shown in table 2.

Obtaining a standard curve regression equation: a is 9.6930C-0.1092, r is 0.9998.

TABLE 2 Absorbance of glucose control at different concentrations

Measuring a sample, precisely measuring 0.6ml of test solution, placing the test solution into a 5ml measuring flask, adding water to dilute the test solution to a scale, shaking the test solution uniformly, precisely measuring 1ml of the test solution, placing the test solution into a 5ml test tube with a plug scale, and measuring absorbance by a method from the point of adding 0.5ml of 4% phenol solution according to the method under the preparation item of a standard curve, and calculating the content; the results are shown in Table 3.

TABLE 3 orthogonal experimental design and result (%) calculation table for ethanol reflux extraction process

(4) Conclusion

Ethanol reflux of Salicornia bigeloviiThe optimal process conditions for extraction are that A₂B₂C₃D₁The combination scheme is that the coarse powder of the medicinal materials is sequentially added with 70% ethanol in an amount which is 10 times, 8 times and 8 times of the coarse powder, extraction is carried out for 3 times, reflux is carried out for 2 hours, 1 hour and 1 hour in sequence, and the extraction amount of the agriophyllum squarrosum oligosaccharide is the highest.

2. Research on macroporous adsorption resin separation and purification process

The aqueous solution of the extract passes through a proper nonpolar or low-polarity macroporous adsorption resin column under a neutral condition, organic lipophilic substances such as flavone, alkaloid, pigment and the like are adsorbed on the macroporous adsorption resin after water elution, and the carbohydrate component has high polarity and cannot be adsorbed on the macroporous adsorption resin, so that the carbohydrate component is further purified.

The substances adsorbed on the macroporous adsorption resin are subjected to gradient elution by ethanol, and different substances can be sequentially eluted according to the polarity of the substances. In the following experiments, by comparing the adsorption and purification performance of several macroporous adsorption resins on the suaeda salsa oligosaccharide, a resin with good separation and purification effects on the suaeda salsa oligosaccharide extract is searched and the separation and purification process thereof is optimized,

preparation method of extract of suaeda salsa

According to the method under the item of the preparation process of the agriophyllum squarrosum oligosaccharide, the agriophyllum squarrosum is taken and crushed into coarse powder, 10 and 8 times of 95 percent ethanol is added in sequence for reflux extraction for 2 times, and each time is 2 hours, and fat-soluble components such as chlorophyll and the like are removed; reflux-extracting defatted medicinal powder with 10, 8, and 8 times of 70% ethanol for 3 times, 2 hr for the first time, 1 hr for the second and third times, filtering the extractive solutions, mixing filtrates, recovering ethanol under reduced pressure, concentrating to no ethanol smell (1g:1ml), diluting with equal volume of water, standing at low temperature overnight, filtering to obtain extractive solution, and refrigerating. Each 1ml contains 0.5g of crude drug. Adding water to dilute to the required concentration;

determination of analytical methods

A phenol-sulfuric acid method or anthrone-sulfuric acid method for quantitatively analyzing sugar is based on that under the action of concentrated sulfuric acid, the sugar is hydrolyzed and dewatered to generate furfural or hydroxymethyl furfural, which is then condensed with phenol to obtain orange-red compound or condensed with anthrone to obtain blue-green derivative, which has color shade in a certain rangeIs proportional and has a maximum absorption at a particular wavelength at which the level is determined colorimetrically. During the content determination process, glycosides such as flavone contained in the crude extract of Salicornia herbacea are hydrolyzed into sugar under the action of concentrated sulfuric acid, and the sugar is further hydrolyzed to interfere with the determination result of oligosaccharide. The content of total flavone is determined by NaNO₂-Al(NO₃)₃-NaOH colorimetry.

Therefore, the test mainly takes the flavonoid components to be separated and removed as the examination index, inspects and determines the total oligosaccharide under the appropriate condition, and comprehensively evaluates the process for preparing the oligosaccharide by separating, purifying and purifying the macroporous adsorption resin.

Screening of the macroporous adsorption resin

7 static adsorption and desorption tests of macroporous adsorption resin

The pretreatment of the macroporous adsorption resin comprises the steps of taking a certain amount of resin to be selected, soaking the resin in 95% ethanol for 24 hours, washing the resin with the ethanol until the eluate is white and turbid, then washing the resin with distilled water until the eluate is alcohol-free, sequentially soaking the resin in 5% HCI and 4% NaOH for 2-4 hours, respectively washing the resin with purified water until the eluate is neutral, and performing suction filtration on the resin by using a Buchner funnel for later use. The physical property parameters of the 7 macroporous adsorption resins to be selected are shown in the table 4:

TABLE 47 physical property parameter tables of macroporous adsorbent resins

Measuring Salicornia herbacea initial extract 40ml under static adsorption amount investigation, diluting with water to 1000ml, shaking up (0.2g crude drug/ml), and determining total flavone content (C)₀). Weighing three parts of macroporous adsorbent resin, each 1g (W), placing in 100mL triangular flask with plug, and accurately adding 40mL (V) of the above crude extract water solution of herba Salicornia Bicoloris₀) Weighing, placing in a shaking table, shaking for 24 hr, weighing again, adding water to supplement loss weight, shaking, filtering, and measuring the content of residual total flavone (C)₁) The resin filter cake is washed with water and the washing liquid is discarded. And calculating the total flavone adsorption quantity (Q) of each macroporous adsorption resin according to the following formula. The results are shown in Table 5.

Static desorption rate examination the macroporous resin adsorbed with the general flavone of Salicornia Herbacea was transferred to a 100mL Erlenmeyer flask with a stopper, and 30mL (V) of 70% ethanol was added₁) Weighing, placing in a shaking table, shaking for 24h, weighing, adding 70% ethanol to complement loss weight, shaking, filtering, and measuring the content C of the residual total flavone in the filtrate₂. And calculating the desorption amount (D) and the desorption rate (E) of the total flavonoids of each macroporous adsorption resin according to the following formulas, and the results are shown in Table 5:

TABLE 57 static adsorption and desorption performance tables for macroporous adsorption resin

(4) Conclusion

The test results shown in table 5 indicate: the static adsorption and desorption capacities of the 7 kinds of macroporous adsorption resin are obviously different, and the best static adsorption capacity and desorption capacity is D101 type, and then AB-8 type; the ADS-7 type adsorption capacity is good, but the desorption rate is poor and the desorption capacity is low.

Therefore, the comprehensive consideration is that 2 kinds of macroporous adsorption resins of D101 type and AB-8 type are selected to continue the dynamic adsorption and desorption tests so as to select the better one.

II, secondly: pharmacological research of the suaeda salsa oligosaccharide on heart injury:

1. salicornia bigelovii oligosaccharide dosage

The clinical application method comprises the following steps: the composition is administered orally at a dose of 0.075g/kg once a day for an adult;

the equivalent dose ratio of rat to human is 6.3:1, so the equivalent dose for rat treatment is 0.47 g/kg. Three doses, 2 times, 1 time (equal time) and 0.5 time the clinical dosage of human, were used in the rat experiment. The dosage is 0.94, 0.47, 0.24g/kg respectively. The administration route is oral administration (gavage), the volume is 5ml/kg, and the administration is carried out once a day at 8: 00-10: 00 am.

2. Establishing a myocardial injury animal model: isoproterenol-induced myocardial injury in rats

Feeding 90 SD rats for 1 week, randomly reserving 10 rats as a normal control group after the adaptation is finished, injecting isoproterenol into the abdominal cavity of the other 80 rats to induce myocardial damage, wherein the dose is 5mg/(kg d), and injecting normal saline with the same dose into the normal control group for 2 weeks continuously; after 2 weeks, 60 rats were injected with isoproterenol and the success of the rat model was evaluated by cardiac ultrasound; the 40 successfully molded rats were randomly divided into 4 groups: 10 model control groups, 0.94g/(kgd)10 Salicornia bigelovii oligosaccharide high dose groups, 0.47g/(kg d)10 Salicornia bigelovii oligosaccharide medium dose groups and 0.24g/(kg d)10 Salicornia bigelovii oligosaccharide low dose groups. The drug treatment groups are respectively used for the intragastric administration treatment according to the drug dosage, the normal control group and the model control group are given distilled water with the same volume once a day, the administration is carried out at 8-10 am, and the continuous administration is carried out for 2 weeks.

3. Grouping and administration of drugs

Rats meeting the modeling standard are randomly divided into 5 groups, namely a normal control group, a model control group, a high-dose group of the agriophyllum squarrosum oligosaccharide, a medium-dose group of the agriophyllum squarrosum oligosaccharide and a low-dose group of the agriophyllum squarrosum oligosaccharide.

4. Observation index

4.1 protective Effect of Salicornia bigelovii oligosaccharide on isoproterenol-induced myocardial injury in rats

4.1.1 detection of myocardial enzymes and antioxidant indexes in rat blood

TABLE 6 Effect of Salicornia bigelovii oligosaccharide on Isoprenin-induced myocardial enzyme levels in rats: (n＝10)，U·L^-1

Note: compared with the normal control group,^#P<0.05，^##P<0.01; compared with the model control group,^*P<0.05，^**P<0.01

table 6 the results show: compared with a normal control group, the serum myocardial enzymology indexes of the model group mice, namely Lactate Dehydrogenase (LDH), Creatine Kinase (CK) and creatine kinase isozyme (CK-MB), are obviously increased (P is less than 0.01); compared with the model group, the administration groups of the agriophyllum squarrosum oligosaccharide obviously reduce the levels of LDH, CK and CK-MB in the serum of rats (P is less than 0.01, and P is less than 0.05).

TABLE 7 Effect of Salicornia bigelovii oligosaccharides on Isoprenin-induced oxidative stress in rats: (n＝10)，U·mg^-1

Note: compared with the normal control group,^#P<0.05，^##P<0.01; compared with the model control group,^*P<0.05，^**P<0.01

table 7 the results show: isoproterenol induces the serum lipid peroxidation level of the rats to be increased to different degrees, wherein the MDA and the GSH-PX in the serum of the rats in the model group are obviously increased and the SOD is obviously reduced (P is less than 0.01 and P is less than 0.05) compared with the normal control group; the high and medium dose of the agriophyllum squarrosum oligosaccharide can reduce the MDA and GSH-PX level in serum and increase the SOD level when being independently applied.

Pathologically, it can be analyzed that:

MDA is a product of lipid peroxidation, the obvious increase of MDA indicates that a cell membrane structure suffers from a large degree of oxidative damage, after isoproterenol enters an organism, myocardial cells are more likely to generate lipid peroxidation, and the MDA content is increased, so the MDA is an index of the severity of cardiac cell damage induced by isoproterenol.

The blank group has obviously increased MDA content in serum after induction by isoproterenol, and has obviously reduced MDA content after treatment by different dosages of Salicornia bigelovii oligosaccharide. Meanwhile, under the induction of isoproterenol, a large amount of myocardial enzyme of the rat can be released, so that the rat is induced to have myocardial injury, and whether myocardial cells have injury and necrosis can be indirectly reflected by detecting the level of the myocardial enzyme. The activities of LDH, CK and CK-MB in the serum of rats in the model group are obviously improved, and the activities of LDH, CK and CK-MB in the serum of rats in each group of the agriophyllum squarrosum oligosaccharide group are obviously reduced.

Therefore, the above results show that the agriophyllum squarrosum oligosaccharide can play a role in protecting the rat myocardial damage induced by the isoprochane epixin by reducing the release of the damaged myocardial enzymes, improving the activities of SOD and GSH-PX in the rat serum and reducing the content of MDA.

Rats can be induced by isoproterenol, so that the heart endogenous antioxidant system is reduced to cause oxidative damage, and the serum lipid peroxidation level of the rats is increased to different degrees. The endogenous antioxidant enzyme system, consisting of SOD, CAT and GSH-Px, is the first cellular defense system against oxidative stress, associated with the formation of antioxidants, including superoxide anions and hydroxyl radicals. The activity of SOD is sharply reduced after isoproterenol stimulation, GSH-PX is obviously increased, and the irresistibility of free radicals is indicated, so that the oxidative damage of myocardial cells is caused.

After induction with isoproterenol in the blank group, SOD was significantly reduced and GSH-PX was significantly increased. After the experimental group is treated by different dosages of the suaeda salsa oligosaccharide, the SOD activity level in the serum of rats induced by isoproterenol is obviously improved, the GSH-PX is obviously reduced,

therefore, the agriophyllum squarrosum oligosaccharide has a certain degree of antioxidant capacity in rat myocardial infarction caused by isoproterenol.

4.1.2 rat Heart/body Mass index

Immediately cutting open the abdomen and taking the heart of each rat after blood is taken, sucking the blood by using filter paper, immediately weighing the heart by using an electronic balance, quickly taking 1-2 cubic millimeters of tissue, putting the tissue into a small bottle containing glutaraldehyde, and observing the ultrastructure by using a transmission electron microscope; fixing part of the tissue in 4% paraformaldehyde (fixing for over 72h, and changing the medium for 2 times), performing conventional paraffin embedding, and observing the structure of the case under HE staining light.

The body weight, heart weight and heart/body mass index pairs for each group of rats are shown in table 8: compared with a normal control group, the weight of the rat in the model group is obviously reduced, the statistical significance is achieved (P is less than 0.05), the heart weight and the heart/body weight index are obviously increased, and the statistical significance is achieved (P is less than 0.05); the weight of the rats in the high, medium and low treatment groups of the agriophyllum squarrosum oligosaccharide is increased compared with the model group, and the heart weight and the heart/body weight index of the rats in the high dose group of the agriophyllum squarrosum oligosaccharide are reduced compared with the model group, wherein the weight, the heart weight and the heart/body weight index of the rats in the high dose group of the agriophyllum squarrosum oligosaccharide have statistical significance (P is less than 0.05) compared with the model group.

TABLE 8 Effect of Salicornia bigelovii oligosaccharides on the isoprenaline-induced Heart/body Mass index in rats: (n＝10)

Note: compared with the normal control group,^#P<0.05，^##P<0.01; compared with the model control group,^*P<0.05，^**P<0.01

4.1.3 changes in the Electrocardiogram of rats

Isoproterenol is a beta-adrenergic receptor agonist, has the function of exciting and exciting a heart beta receptor, can enhance the cardiac contractility, accelerate the heart rate and increase the cardiac output when continuously applied in large dose, can simultaneously cause the oxygen consumption of myocardial cells to be greatly increased, generate myocardial energy metabolism disorder, cause myocardial ischemia and hypoxia, further cause abnormal change of the S-T section of an electrocardiogram of a rat, generate myocardial injury necrosis, generate electrocardiogram change when the large dose of isoproterenol induces myocardial injury of animals, serological change and myocardial histopathological change which are very similar to the change of human beings when myocardial infarction occurs,

therefore, whether the normal electrophysiological activity of the heart is affected can be detected through electrocardiographic examination, whether the organic and functional lesions of the heart are generated or not is presumed, as shown in fig. 1-6, the heart rate of a normal control group is normal before the normal saline is given, the heart rate of a rat is slightly increased after the normal saline is given, other factors have no obvious influence, after the rat is molded by isoproterenol, the S-T section of the electrocardiogram of the rat is raised, the T wave is flat, the heart rate is obviously accelerated, and the conditions of myocardial ischemia and tachycardia are indicated; compared with the electrocardiogram of a rat in a high and medium dose treatment group of the suaeda salsa oligosaccharide, the electrocardiogram S-T section is reduced, the T wave is increased, and the heart rate is obviously reduced; compared with the model group, the electrocardiogram of the rats in the low-dose treatment group of the suaeda salsa oligosaccharide has no obvious change, but the heart rate is slightly reduced;

the data of the invention show that the high and medium dose groups of the suaeda salsa oligosaccharide can inhibit the conditions of elevation of the S-T section of the electrocardiogram of a rat caused by isoproterenol, low and flat T wave and obviously accelerated heart rate, which indicates that the suaeda salsa oligosaccharide can improve the conditions of myocardial ischemia and tachycardia.

4.1.4 pathological changes in rat Heart

As shown in fig. 7, wherein: a: normal group B: model group C: sarabi oligosaccharide high dose group D: sarabi oligosaccharide medium dose group E: low dose group of suaeda salsa oligosaccharide;

the results of the figures show that: the normal control group has complete myocardial tissue cells, normal structure, clear boundary, no pathological changes such as hypertrophy, degeneration, necrosis and the like, uniform cytoplasmic staining and nucleus ellipse in the cells. Compared with the blank group, the rat myocardial tissue cells of the model group are disintegrated and necrotic, the cells are disorderly arranged and have unclear boundaries, the cell nucleuses are scattered, diffuse necrosis is formed, the cytoplasm is turbid, and the cell nucleuses are scattered among the cells; compared with the model group, the rat myocardial tissue cells of the high, medium and low dose treatment groups of the agriophyllum squarrosum oligosaccharide are arranged in order, the lytic necrosis condition is lighter compared with that of the model group, the improvement condition of the high dose group of the agriophyllum squarrosum oligosaccharide is the best, the treatment condition is dose-dependent in the medium dose group of the agriophyllum squarrosum oligosaccharide and in the low dose group of the agriophyllum squarrosum oligosaccharide,

the Salicornia bigelovii oligosaccharide dosage groups can improve rat myocardial injury caused by isoproterenol, so that the myocardial cell morphological structure tends to be complete, the arrangement is compact and neat, the boundary is clear, the cytoplasm is uniformly dyed, the nucleus is clear, and the cytoplasm is positioned in the center of the cell.

From the above analysis, it can be seen that:

(1) the Salicornia bigelovii oligosaccharide is shown to improve necrotic myocardial cells according to histopathology, and improve acute myocardial injury caused by coronary artery lesion caused by isoproterenol induced cell necrosis ischemia by maintaining the integrity of the myocardial cells;

(2) the suaeda salsa oligosaccharide can improve and relieve myocardial ischemia and tachycardia, and improve acute myocardial injury caused by isoproterenol-induced myocardial ischemia and tachycardia according to an electrocardiogram;

(3) the agriophyllum squarrosum oligosaccharide can also improve the activities of SOD and GSH-PX in rat serum and reduce the content of MDA by reducing the release of damaged cardiac myozyme, thereby playing a role in protecting the rat cardiac muscle damage induced by isoproterenol and improving the cardiac muscle damage of isoproterenol induced hormone level;

therefore, the Salicornia bigelovii oligosaccharide can effectively relieve the problem that the cardiac muscle enzyme and the antioxidant index are obviously increased due to isoproterenol, and the myocardial damage of myocardial stress and body stress is caused.

In conclusion: the agriophyllum squarrosum oligosaccharide has good effect on the induction of myocardial injury by various pathological mechanisms of isoproterenol, has certain improvement effect on the myocardial injury of rats caused by various pathological mechanisms of isoproterenol, and has certain protection effect on the myocardial injury.