Application of Shexiangxintongning in preparing medicine for treating ischemic stroke

Abstract

The present invention belongs to the field of medical technology, and specifically relates to the use of Musk Xintongning in the preparation of drugs for treating ischemic stroke. The present invention reports for the first time the efficacy of Musk Xintongning in treating ischemic stroke. Animal experimental results show that Musk Xintongning tablets can greatly reduce the area of ​​cerebral infarction lesions, inhibit neuronal cell necrosis, and significantly improve the neurobehavior and cognitive level of MCAO rats. The present invention is of great significance for the use of Musk Xintongning in treating ischemic stroke.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and particularly relates to the application of Musk Heart Tong Ning in preparing a medicine for treating ischemic stroke. Background Art

[0002] Ischemic stroke, also known as cerebral infarction, is called stroke or apoplexy in traditional Chinese medicine. The disease is caused by various reasons, resulting in obstruction of blood supply and ischemia in brain tissue, leading to a syndrome of ischemic hypoxic necrosis and softening of brain tissue, thus producing clinically corresponding neurological defects, mainly manifested as sudden focal or diffuse neurological deficits.

[0003] In current medicine, ischemic stroke has the characteristics of high incidence, high recurrence rate, high disability rate and high mortality rate. Its prognosis is related to the severity of neurological deficit, age and etiology. If it can be treated in time, some patients may recover some motor and sensory functions. The emergence of this disease has caused invisible harm to human health.

[0004] Currently, commonly used drugs for the treatment of ischemic stroke include drugs that dilate blood vessels, expand blood volume, dissolve blood clots, or anticoagulants. However, blood volume expander drugs are not suitable for people with heart disease, as they may cause heart failure once taken. Thrombolytic drugs should not be taken in large quantities as they may cause bleeding in the body. When taking anticoagulants, special attention should be paid to adverse reactions.

[0005] The main ingredients of Musk Heart Pain Relief Tablets are artificial musk, Corydalis yanhusuo (fried with vinegar), storax, Chuanxiong, ginseng, and borneol, and are specifically made of 220-260 parts of Corydalis yanhusuo (fried with vinegar), 4-12 parts of artificial musk, 4-12 parts of storax, 100-140 parts of Chuanxiong, 100-140 parts of ginseng, and 6-18 parts of borneol. Currently, it is used to treat chest pain, chest tightness, distending pain in both flanks, shortness of breath, palpitations and other symptoms caused by angina pectoris of coronary heart disease of qi stagnation and blood stasis type, and its use in the treatment of ischemic stroke has not been reported. Summary of the invention

[0006] In order to solve the above problems, the present invention provides the use of Musk Heart Tong Ning in preparing a medicine for treating ischemic stroke.

[0007] The present invention provides the following technical solutions:

[0008] Application of Musk Heart Tong Ning in the preparation of drugs for treating cerebral stroke.

[0009] Furthermore, the stroke is an ischemic stroke.

[0010] Furthermore, the drug is in any pharmaceutically acceptable dosage form, including at least one of tablets, capsules, injections, granules, suspensions and solutions.

[0011] Furthermore, the preparation method of the above Musk Heart Tong Ning is as follows:

[0012] The following six herbs: 239g of vinegar-fried Corydalis, 8g of artificial musk, 8g of storax, 119g of Chuanxiong, 119g of ginseng, 12g of borneol, artificial musk plus an equal amount of starch, mixed and ground into fine powder; storax plus 12g of micro-powdered silica gel, mixed and ground into fine powder; borneol ground into fine powder. 80g of Corydalis (vinegar-fried) is crushed into fine powder; the remaining 159g of Corydalis (vinegar-fried), ginseng, and Chuanxiong are refluxed and extracted three times with 60% ethanol, each time adding 6 times the amount, refluxed for 2 hours, the extracts are combined, filtered, the filtrate is used to recover ethanol and concentrated to a thick paste with a relative density of 1.30-1.32 (60°C), Corydalis (vinegar-fried) fine powder and starch in appropriate amounts are added, mixed, granulated, dried, artificial musk, storax, and borneol fine powder are added, mixed, pressed into 1000 tablets, and film-coated to obtain the product.

[0013] The beneficial effects of the present invention include but are not limited to:

[0014] The present invention reports for the first time the application of Musk Xintongning in the treatment of ischemic stroke. The results of animal experiments show that the administration of Musk Xintongning tablets can greatly reduce the area of ​​cerebral infarction lesions and inhibit the necrosis of neuronal cells. Moreover, compared with the model group, the novel object preference index of the Musk Xintongning group was significantly increased (P<0.05), indicating that Musk Xintongning tablets can significantly improve the neurobehavior and cognitive level of MCAO rats.

[0015] Through the study on the rationality of the formula of Musk Heart Pain Ning, it is shown that the effect of the whole formula in treating ischemic stroke is the result of the combined action of the main drug, the minister drug and the adjuvant drug, which provides a research basis for the application of Musk Heart Pain Ning in the preparation of drugs for the treatment of ischemic stroke. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] The drawings described herein are used to provide a further understanding of the present invention and constitute a part of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation of the present invention. In the drawings:

[0017] Figure 1 The effect of Shexiangxintongning on the cerebral infarction area in MCAO rats;

[0018] Figure 2 To study the effect of Shexiang Xintongning Tablets on neuronal necrosis in MCAO rats;

[0019] Figure 3 , Figure 4 They are Figure 1 and Figure 2 Color view of . DETAILED DESCRIPTION

[0020] In order to clearly illustrate the technical features of this solution, the present invention is described in detail below through specific implementation methods and in conjunction with the accompanying drawings. The scope of the present invention is not limited to the following embodiments. Professionals in the field can understand that various changes and modifications can be made to the present invention without departing from the spirit and scope of the present invention.

[0021] Experimental application of Shexiang Xintongning tablets in the treatment of ischemic stroke

[0022] 1. Materials

[0023] 1.1 Experimental animals

[0024] SPF male SD rats, 280 g ± 20 g, were purchased from Beijing Weitonglihua Laboratory Animal Technology Co., Ltd., with the experimental animal license number SCXK (Beijing) 2016-0011.

[0025] 1.2 Main instruments

[0026] IVC independent ventilation cage system (BCR-RI01-25-C12 / PSU, Shandong Xinhua Medical Instrument Co., Ltd.), multi-channel small animal anesthesia machine (R550IE, Shenzhen Reward Life Science Co., Ltd.), Morris water maze (Shenzhen Reward Life Science Co., Ltd.), cross elevated rack (63009, Shenzhen Reward Life Science Co., Ltd.), new object recognition (63008, Shenzhen Reward Life Science Co., Ltd.).

[0027] 1.3 Main drugs and reagents

[0028] Bolus (2838 / A4, Beijing Xinong Technology Co., Ltd.), isoflurane (Shenzhen Ruiwode Life Science Co., Ltd., batch number: 20220101), Musk Xintongning (Shandong Hongjitang Pharmaceutical Group Co., Ltd., batch number: 2102001).

[0029] 1.4 Product manufacturing method

[0030]

Ingredients

[0031] [Preparation method] The above six ingredients, artificial musk, add an equal amount of starch, mix and grind; add 12g of micro-powdered silica gel to storax, mix and grind; grind borneol. 80g of Corydalis (fried with vinegar) is crushed into fine powder; the remaining 159g of Corydalis (fried with vinegar), ginseng, and Chuanxiong are refluxed and extracted three times with 60% ethanol, each time adding 6 times the amount, reflux for 2 hours, combine the extracts, filter, recover ethanol from the filtrate and concentrate to a thick paste with a relative density of 1.30-1.32 (60℃), add Corydalis (fried with vinegar) fine powder and starch in appropriate amount, mix well, granulate, dry, add artificial musk, storax and borneol fine powder, mix well, press into 1000 tablets, and coat with film.

[0032] 2. MCAO model establishment

[0033] Rats were anesthetized with isoflurane, and the neck skin was prepared and disinfected. A 2-cm longitudinal incision was made in the middle of the neck, and the neck muscles were bluntly separated to expose and separate the right common carotid artery (CCA), internal carotid artery (ICA), external carotid artery (ECA) and vagus nerve. Wires were hung at the distal and proximal ends of the CCA for use, the distal end of the CCA was clamped with an artery clamp, and the proximal end of the CCA was ligated. A 0.2-mm "V"-shaped incision was made 4-5 mm away from the bifurcation of the CCA, and the suture was slowly inserted into the CCA through the incision. The suture was slowly sent along the ICA into the proximal middle cerebral artery. The suture was stopped when the special black mark on the suture just reached the bifurcation of the ICA and ECA and the suture encountered resistance; the suture was fixed at the distal end of the CCA to prevent it from slipping, the excess suture outside the blood vessel was cut off, and the muscles and skin were sutured; the sham operation group was only given vascular stripping.

[0034] 3. Pharmacological research methods

[0035] 3.1 Animal grouping, medication and treatment

[0036] The animals were scored for behavior 30 minutes after awakening, and randomly divided into the model group and the Musk Heart Pain Ning group, with 8 animals in each group, and the drugs were administered 24 hours later. Musk Heart Pain Ning was administered intragastrically at a raw drug dosage of 0.135g / kg.d, diluted with 5% Tween 80. Another 8 animals were selected as the sham operation group. The model group and the sham operation group were given normal saline for intragastrical administration. The behavioral scores were performed again three weeks after the medication, and 6 animals were randomly selected from the model group, Musk Heart Pain Ning group, and the sham operation group for heart perfusion and brain removal.

[0037] 3.2 Behavioral testing

[0038] Each group was scored weekly according to the modified mNSS neurological deficit scoring standard, and the cross elevated maze was used to test the rats' short-term memory at the end of the first week, and the cross elevated maze was used again at the end of the second week. The novel object recognition experiment was performed every other day to test the rats' learning and memory abilities. Starting from the third week, the Morris water maze was performed for five consecutive days to test the rats' spatial memory abilities.

[0039] 3.3TTC staining

[0040] The rat brain was frozen at -80℃ for about 2 hours, and 4 coronal brain slices were cut from the frontal pole, about 1.5 mm thick, placed in 2% TTC solution, and incubated at 37℃ in the dark for 30 minutes. The ischemic area was stained white, and the non-ischemic area was stained red.

[0041] 3.4 Brain neuron necrosis rate

[0042] Routine 5 μm coronal sections were cut, dewaxed with xylene, washed with gradient alcohol to water, stained with hematoxylin, washed with concentrated ammonia to blue, stained with eosin, and sealed with neutral gum. Positive cells were defined as those with densely stained, condensed, and fragmented nuclei. The rat cerebral cortex and the CA1 region of the ischemic hippocampus were observed under a high-power microscope (40×).

[0043] IV. Pharmacological Study Results

[0044] 4.1 Effects of Shexiang Xintongning Tablets on Cerebral Infarction and Neuronal Necrosis in MCAO Rats

[0045] The results are as follows Figure 1 As shown by Figure 1 It can be seen that the mice in the model group had larger cerebral infarction lesions than those in the sham operation group, and there were a large number of necrotic neuronal cells in the hippocampus and cortex areas. The administration of Musk Heart Pain Ning Tablets can greatly reduce the area of ​​cerebral infarction lesions and inhibit the necrosis of neuronal cells.

[0046] 4.2 Effects of Musk Xintongning Tablets on Neurobehavioral Scores and New Foreign Body Recognition in MCAO Rats

[0047] Combined with Table 1 and Figure 2 Compared with the sham operation group, the neurobehavioral score of the model group was significantly increased (P < 0.01), and compared with the model group, the neurobehavioral score of the Musk Xintongning group was significantly decreased (P < 0.01); compared with the sham operation group, the novel object preference index of the model group was significantly decreased (P < 0.05), and compared with the model group, the novel object preference index of the Musk Xintongning group was significantly increased (P < 0.05). This shows that Musk Xintongning tablets can significantly improve the neurobehavioral and cognitive functions of MCAO rats.

[0048] Table 1 Effects of Shexiang Xintongning Tablets on neurobehavioral scores and new foreign body recognition in MCAO rats

[0049]

[0050] Note: a Compared with the sham operation group, P<0.01, b Compared with the model group, P<0.01; #Compared with the sham operation group, P<0.05, *Compared with the model group, P<0.05.

[0051] 4.3 Effects of Musk Xintongning Tablets on Cross-elevation in MCAO Rats

[0052] Cross-elevated: Compared with the sham operation group, the percentage of open wall time in the model group was significantly reduced (P < 0.05), and compared with the model group, the percentage of open wall time in the Musk Heart Tong Ning group was significantly increased (p < 0.01). Explanation: Musk Heart Tong Ning tablets can significantly improve the anxiety state of MACO rats. The results are shown in Table 2.

[0053] Table 2 Effects of Shexiang Xintongning Tablets on the Cross Elevation of MCAO Rats

[0054]

[0055] Note: a Compared with the sham operation group, P<0.05, b Compared with the model group, P<0.01.

[0056] 4.4 Effects of Musk Xintongning Tablets on Water Maze Test in MCAO Rats

[0057] Water maze: Compared with the sham operation group, the percentage of time on the platform in the model group was significantly reduced (P < 0.01), and compared with the model group, the percentage of time on the platform in the Musk Xintongning group was significantly increased (P < 0.05); compared with the sham operation group, the number of times the model group crossed the platform was significantly reduced (P < 0.01), and compared with the model group, the number of times the Musk Xintongning group crossed the platform was significantly increased (P < 0.01). Explanation: Musk Xintongning tablets can significantly improve the cognitive level of MACO rats. The results are shown in Table 3.

[0058] Table 3 Effects of Shexiang Xintongning Tablets on the Water Maze of MCAO Rats

[0059]

[0060] Note: a Compared with the sham operation group, P<0.01, b Compared with the model group, P < 0.05; # Compared with the sham operation group, P<0.01, * Compared with the model group, P<0.01.

[0061] V. Research on the rationality of the formula:

[0062] The prescription of Musk Heart Pain Ning Tablets was divided into the main drug group (musk, Corydalis yanhusuo), the auxiliary drug group (storax, Chuanxiong), the adjuvant drug group (borneol, ginseng), the main drug group + the auxiliary drug group (musk, Corydalis yanhusuo, storax, Chuanxiong), the main drug group + the adjuvant drug group (musk, Corydalis yanhusuo, borneol, ginseng), the auxiliary drug group + the adjuvant drug group (storax, Chuanxiong, borneol, ginseng), and the tablets were made according to the original prescription process. The MCAO rat model was made, and the effects of each group on the rat neurobehavioral score and new foreign body recognition were observed. The results are shown in Table 4.

[0063] Compared with the sham operation group, the neurobehavioral score of the model group was significantly increased (P<0.01), and there was no significant difference in the neurobehavioral score of the monarch drug group, minister drug group, and adjuvant drug group compared with the model group; compared with the sham operation group, the new object preference index of the model group was significantly reduced (P<0.05), and there was no significant change in the new object preference index of the monarch drug group, minister drug group, and adjuvant drug group compared with the model group. Explanation: The effect of Shexiang Xintongning Tablets in treating ischemic stroke is the result of the combined action of monarch drug, minister drug, and adjuvant drug.

[0064] Table 4 Effects of monarch drug group, minister drug group and adjuvant drug group on neurobehavioral scores and new foreign body recognition in MACO rats

[0065]

[0066] Note: a Compared with the sham operation group, P < 0.01; # Compared with the sham operation group, P<0.05.

[0067] In order to investigate the effect of each drug in the prescription, a musk negative sample (the prescription is the same as the musk xintongning tablets except that there is no musk), a corydalis negative sample (the prescription is the same as the musk xintongning tablets except that there is no corydalis), a storax negative sample (the prescription is the same as the musk xintongning tablets except that there is no storax), a chuanxiong negative sample (the prescription is the same as the musk xintongning tablets except that there is no chuanxiong), a ginseng negative sample (the prescription is the same as the musk xintongning tablets except that there is no ginseng), and a borneol negative sample (the prescription is the same as the musk xintongning tablets except that there is no borneol). MCAO rat models were made to observe the effects of each group on the rat neurobehavioral score and new foreign body recognition. The results are shown in Table 5.

[0068] Table 5 Effects of the above experimental drug groups on neurobehavioral scores and new foreign body recognition in MACO rats

[0069]

[0070]

[0071] Note: aCompared with the sham operation group, P<0.01; #Compared with the sham operation group, P<0.05; bCompared with the model group, P<0.01; *Compared with the model group, P<0.05.

[0072] As can be seen from Table 5, compared with the sham operation group, the neurobehavioral score of the model group was significantly increased (P < 0.01); compared with the model group, the neurobehavioral scores of the musk negative group, the corydalis negative group, the storax negative group, the chuanxiong negative group, the ginseng negative group, and the borneol negative group all showed a certain degree of decrease; compared with the sham operation group, the new object preference index of the model group was significantly reduced (P < 0.05); compared with the model group, the new object preference index of the musk negative group, the corydalis negative group, the storax negative group, the chuanxiong negative group, the ginseng negative group, and the borneol negative group all changed significantly. This shows that the Musk Xintongning Tablet has the effect of treating ischemic stroke as a result of the combined action of each raw material, and none of them can be missing.

[0073] The above is only the embodiment of the present application, and the protection scope of the present application is not limited by these specific embodiments, but is determined by the claims of the present application. For those skilled in the art, the present application can have various changes and variations. Any modification, equivalent replacement, improvement, etc. made within the technical ideas and principles of the present application should be included in the protection scope of the present application.

Claims

1. Application of Musk Heart Tong Ning in the preparation of a drug for treating stroke, wherein the Musk Heart Tong Ning comprises 239 g of vinegar-fried Corydalis yanhusuo, 8 g of artificial musk, 8 g of storax, 119 g of Chuanxiong, 119 g of ginseng, and 12 g of borneol, and the preparation method is as follows: For the above six herbs, artificial musk is mixed with an equal amount of starch and ground into fine powder; storax is mixed with 12g of micro-powdered silica gel and ground into fine powder; borneol is ground into fine powder; 80g of vinegar-roasted Corydalis yanhusuo is crushed into fine powder; the remaining 159g of vinegar-roasted Corydalis yanhusuo, ginseng and Chuanxiong are refluxed and extracted three times with 60% ethanol, each time adding 6 times the amount, refluxed for 2 hours, the extracts are combined, filtered, ethanol is recovered from the filtrate and concentrated to a thick paste with a relative density of 1.30-1.32 at 60°C, and an appropriate amount of vinegar-roasted Corydalis yanhusuo powder and starch are added, mixed evenly, granulated, dried, and artificial musk, storax and borneol powder are added, mixed evenly, and the paste is obtained.

2. The use according to claim 1, characterized in that: Stroke is an ischemic stroke.

3. The use according to claim 1, characterized in that: The drug is in any pharmaceutically acceptable dosage form, including at least one of tablets, capsules, injections, granules, suspensions and solutions.

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