Use of Shouhui Tongbian Capsule in the preparation of a medicament for preventing or treating ischemic stroke

Abstract

The present invention provides the use of a traditional Chinese medicine composition prepared from polygonum multiflorum, aloe, cassia seed, wolfberry fruit, donkey-hide gelatin, ginseng, atractylodes macrocephala and fructus aurantii immaturus as main components in the preparation of a drug for preventing or treating ischemic stroke. The research results show that Shouhui Laxative Capsule can reduce the neurological deficit score of cerebral ischemia-reperfusion injury, decrease the area of cerebral infarction, relieve cerebral pathological injury, and reduce the inflammatory level in serum, and has a significant brain protection effect on cerebral ischemia-reperfusion injury. The traditional Chinese medicine composition of the present invention has definite curative effect and safe components, and has broad application prospects and positive clinical significance.

Description

Technical Field

[0001] The present invention belongs to the field of medicine and relates to the use of Shouhui Laxative Capsule in the preparation of a medicament for preventing or treating ischemic stroke. Background Art

[0002] Ischemic cerebrovascular disease (ICVD) refers to the degeneration, necrosis or transient loss of function of local brain tissue due to blood supply disorders. It is one of the common diseases of the nervous system, which is prone to occur in middle-aged and elderly people, and has a relatively high disability rate and fatality rate. The causes of ischemic cerebrovascular disease include arteriosclerosis, vasculitis, congenital vascular diseases, trauma, drugs, blood diseases, etc. Clinical common symptoms include unilateral facial numbness or limb numbness, weakness, language disorders, dizziness accompanied by vomiting, severe headache that was rarely seen in the past, disturbance of consciousness or convulsions, etc.

[0003] "Classification of Cerebrovascular Diseases in China 2015" classifies ischemic cerebrovascular diseases into the following types: transient ischemic attack, cerebral infarction (acute ischemic stroke), cerebral artery steal syndrome, and chronic cerebral ischemia.

[0004] Stroke, abbreviated as "zhongfeng", is a common acute cerebrovascular disease, including ischemic stroke and hemorrhagic stroke. The incidence of ischemic stroke is much higher than that of hemorrhagic stroke, accounting for up to 60% - 70%. Thrombolytic therapy is currently recognized as the most effective treatment method clinically, but it has strict requirements for the treatment time window, and it will cause secondary brain injury after thrombolysis, that is, cerebral ischemia / reperfusion injury (CIRI), which is most common in focal cerebral ischemia. Severe cases can lead to death or disability. After sudden cerebral ischemia / reperfusion injury, a series of pathological processes such as nerve cell necrosis, apoptosis, blood-brain barrier damage, and inflammatory reactions will occur, leading to metabolic disorders of the body through the peripheral circulation, damage to the intestinal barrier, and accumulation of harmful toxins such as inflammatory factors, which will further aggravate brain dysfunction.

[0005] Traditional Chinese medicine believes that focal cerebral ischemia belongs to the category of "stroke". "The brain is the residence of the original spirit" (Compendium of Materia Medica), which is the pivot of life and dominates the life activities of the human body.

[0006] Shouhui Laxative Capsule is composed of fleece-flower root, aloe, cassia seed, donkey-hide gelatin, Chinese wolfberry, ginseng, atractylodes macrocephala, and immature bitter orange, etc. In the formula, ginseng greatly tonifies primordial qi and promotes the production of body fluid; aloe clears liver heat; fleece-flower root and donkey-hide gelatin nourish yin and moisten dryness, and nourish blood and moisten the intestine; cassia seed, Chinese wolfberry, and atractylodes macrocephala purge heat and relieve constipation, tonify qi and nourish yin to moisten the intestine; immature bitter orange breaks qi and reduces accumulation, and serves as a guiding drug; the whole formula nourishes yin and tonifies qi, and combines tonification and purgation, and can improve the clinical symptoms such as difficult defecation in patients with functional constipation by regulating gastrointestinal hormones.

[0007] Chinese Patent CN100453105C discloses a composition with functions of promoting defecation and detoxification, reducing weight and lipid, and a preparation method thereof. The composition is prepared from eight traditional Chinese medicines, namely Polygonum multiflorum Thunb., Aloe vera L., Cassia obtusifolia L., Lycium barbarum L., Colla Corii Asini, Panax ginseng C. A. Mey., Atractylodes macrocephala Koidz., and Aurantii Fructus Immaturus, and can be used for treating functional constipation; Chinese Patent CN111228389A discloses the use of Shouhui Laxative Capsule in the preparation of drugs for preventing and treating abdominal distension. At present, there is no report on the improvement of ischemic stroke by Shouhui Laxative Capsule. Summary of the Invention

[0008] The present invention provides a use of a traditional Chinese medicine composition in the preparation of a drug for preventing or treating ischemic stroke, and the traditional Chinese medicine composition is prepared from Polygonum multiflorum Thunb., Aloe vera L., Cassia obtusifolia L., Lycium barbarum L., Colla Corii Asini, Panax ginseng C. A. Mey., Atractylodes macrocephala Koidz., and Aurantii Fructus Immaturus as main components.

[0009] Furthermore, the traditional Chinese medicine composition is prepared from the following parts by weight of Polygonum multiflorum Thunb., Aloe vera L., Cassia obtusifolia L., Lycium barbarum L., Colla Corii Asini, Panax ginseng C. A. Mey., Atractylodes macrocephala Koidz., and Aurantii Fructus Immaturus as main components:

[0010] Polygonum multiflorum Thunb. 60 - 150 parts by weight, Aloe vera L. 100 - 200 parts by weight, Cassia obtusifolia L. 80 - 180 parts by weight, Lycium barbarum L. 30 - 150 parts by weight, Colla Corii Asini 30 - 150 parts by weight, Panax ginseng C. A. Mey. 20 - 100 parts by weight, Atractylodes macrocephala Koidz. 20 - 100 parts by weight, Aurantii Fructus Immaturus 50 - 200 parts by weight.

[0011] Even further, the traditional Chinese medicine composition is prepared from the following parts by weight of Polygonum multiflorum Thunb., Aloe vera L., Cassia obtusifolia L., Lycium barbarum L., Colla Corii Asini, Panax ginseng C. A. Mey., Atractylodes macrocephala Koidz., and Aurantii Fructus Immaturus as main components:

[0012] Polygonum multiflorum Thunb. 120 parts by weight, Aloe vera L. 160 parts by weight, Cassia obtusifolia L. 140 parts by weight

[0013] Lycium barbarum L. 75 parts by weight, Colla Corii Asini 75 parts by weight, Panax ginseng C. A. Mey. 50 parts by weight

[0014] Atractylodes macrocephala Koidz. 50 parts by weight, Aurantii Fructus Immaturus 120 parts by weight.

[0015] The traditional Chinese medicine composition provided by the present invention can be prepared into a clinically acceptable dosage form by adding pharmaceutically acceptable excipients, such as oral preparations or injection preparations.

[0016] Furthermore, the dosage form can be pills, tablets, granules, capsules, oral liquids, suspensions, mixtures, powders, injections.

[0017] The drug of the present invention is Shouhui Laxative Capsule.

[0018] The present invention is for the development of new uses of Shouhui Laxative Capsules. The patent for the Shouhui Laxative Capsule product is CN100453105C, and the invention title "A Composition with Laxative and Detoxifying, Weight Loss and Lipid-lowering Functions and Its Preparation Method" has been authorized. The traditional Chinese medicine composition described in the present invention is prepared according to the preparation or extraction methods of each component in patent CN100453105C.

[0019] The present invention also provides the use of the traditional Chinese medicine composition in the preparation of drugs for preventing or treating ischemic cerebrovascular diseases.

[0020] In one embodiment of the present invention, the ischemic cerebrovascular disease is ischemic stroke.

[0021] In one embodiment of the present invention, the ischemic cerebrovascular disease is cerebral ischemia-reperfusion injury.

[0022] Further, in one embodiment of the present invention, the traditional Chinese medicine composition reduces the neurological deficit score of ischemic stroke and promotes the recovery of neurological function.

[0023] Further, in one embodiment of the present invention, the traditional Chinese medicine composition reduces the cerebral infarction area of ischemic stroke.

[0024] Further, in one embodiment of the present invention, the traditional Chinese medicine composition alleviates the pathological damage of the brain in ischemic stroke.

[0025] The present invention has achieved the following technical effects:

[0026] The present invention uses the suture method to establish a rat model of cerebral ischemia-reperfusion injury (CIRI), evaluates the neurological impairment of rats through the Zea-longa grade neurological function scoring table, observes the pathological damage of rat brain tissue by HE staining, and measures the cerebral infarction volume of rats by TTC staining. The experimental results show that Shouhui Laxative Capsules reduce the neurological deficit score of cerebral ischemia-reperfusion injury, promote the recovery of neurological function, reduce the cerebral infarction area, alleviate the pathological damage of the brain, and reduce the inflammatory level in the serum, and have a significant brain protection effect on cerebral ischemia-reperfusion injury. Description of the Drawings

[0027] Figure 1 For the percentage of cerebral infarction volume and TTC staining of rats in each group (x±s, n = 3).

[0028] Figure 2 For observing the pathological conditions of rat brain tissue in each group by HE staining.

[0029] Figure 3To detect the expression of inflammatory factors in the serum of rats in each group after cerebral ischemia-reperfusion by ELISA (x±s, n = 6). Detailed implementation mode

[0030] The present invention will be further described in detail below with reference to specific embodiments. The following embodiments give detailed implementation modes and specific operation processes to illustrate the technical solutions of the present invention, but do not limit the protection scope of the present invention. Those of ordinary skill in the art who modify or equivalently replace the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention still fall within the protection scope of the present invention.

[0031] Example 1 Protective effect of Shouhui Tongbian Capsule on rats with cerebral ischemia-reperfusion model

[0032] 1. Materials and methods

[0033] 1.1 Animals

[0034] 60 male SD rats, weighing 180 - 200 g, were purchased from Beijing Huafukang Biotechnology Co., Ltd., and the production license number is SCX(Beijing)2019 - 0008.

[0035] 1.2 Reagents and instruments

[0036] Shouhui Tongbian Capsule (Lunan Houpu Pharmaceutical Co., Ltd., national drug approval number Z20150041); edaravone (Kunming Jida Pharmaceutical Co., Ltd., national drug approval number H20080495); TNF-α, IL-6, IL-1β ELISA kits (Shanghai Enzyme-linked Biotechnology Co., Ltd., batch numbers m1002859, m102828, m1037361); 2,3,5-triphenyltetrazolium chloride (TTC) (sigma aldrich company, T8877 - 25G). ELX-800 type microplate reader (Biotek company, USA); automatic stainer (leica biosystems nussloch Gmbh company, Germany, ST5020).

[0037] 1.3 Animal grouping and model establishment

[0038] The 60 rats were randomly divided into a sham operation group, a model group, a low-dose group of Shouhui Tongbian Capsule (SHTB) (0.2 g·kg -1 .d -1 ), and a high-dose group of Shouhui Tongbian Capsule (SHTB) (0.4 g·kg -1· d -1) Positive drug group (edaravone injection at 5.4 mg / kg), with 12 rats in each group. According to the body surface area calculation method, the drug dosage for rats is about 6 times that of the human dosage. Therefore, the SHTB dosage in this experiment is planned to be 0.2 g·kg -1 ·d -1 、0.4 g·kg -1 ·d -1 as the low and high dosages. According to the body weight of the rats, 15 mg·mL -1 of Zoletil 50 and 15 mg·mL -1 of xylazine hydrochloride mixture were used to anesthetize the rats at 5 mL·kg. The right common carotid artery (CCA) was isolated and exposed. A suture was pre-placed outside the blood vessel. Continuing to separate and expose the external carotid artery (ECA) upward along the CCA, the proximal ends of the CCA and ECA were ligated with sutures. The distal end of the CCA was clamped with an artery clip. A small incision was made 4 mm from the bifurcation of the CCA. After inserting a 0.24-mm suture, a slipknot was made with the suture. The artery clip was removed. The ECA was slightly lifted and the suture was slowly inserted until resistance was felt to block the blood vessel. The suture at the distal end of the CCA was tightly tied, and the tail end of the suture was exposed outside. After 1.5 h, the suture was withdrawn, and the cerebral ischemia-reperfusion injury (CIRI) model was completed. The wound was sutured. In the sham operation group, except for not inserting the suture, the other steps were the same as those in the model group. The body temperature was maintained at 36.5°C - 37.5°C during the operation. Seven days before modeling, the rats in each group were given intragastric administration until 24 h after modeling. The sham operation group and the model group were given intragastric administration with an equal volume of distilled water, and the positive drug group was given intravenous injection.

[0039] 1.4 Neurological deficit score (Longa score) of rats in each group

[0040] After reperfusion for 24 h, behavioral scoring was performed with reference to the Zea-longa grade score:

[0041] 0 point, normal performance;

[0042] 1 point, the forepaw of the rat cannot be fully extended;

[0043] 2 points, the rat rotates in place;

[0044] 3 points, the rat walks and topples to the paralyzed side;

[0045] 4 points, unable to move independently and completely lose consciousness.

[0046] A rat model with a score of 1 - 3 is a successful model.

[0047] 1.5 TTC staining detection of rats in each group

[0048] The brain tissue was taken and cut into 2-mm coronal sections, which were incubated with 2% TTC solution at 37 °C for 20 min and fixed with 4% paraformaldehyde overnight. The non-stained area was the infarct area. The tissue section images were taken with a digital camera and then analyzed with ImageJ software.

[0049] Calculation of infarct volume: Infarct rate % = infarct area / total brain area × 100%.

[0050] 1.6 Pathological changes in the brain tissues of rats in each group

[0051] The brain tissues of rats were fixed with 4% paraformaldehyde, and the paraffin-embedded brain tissues were cut into 5-μm sections. Then, the paraffin sections were successively dewaxed to water with xylene and gradient ethanol, stained with HE, and sealed. The pathological changes were observed under a microscope.

[0052] 1.7 Levels of inflammatory factors in the sera of rats in each group

[0053] After the rats were anesthetized, the abdominal cavity was opened, and blood was taken from the abdominal vena cava and placed in a biochemical tube containing separating gel. After standing for 30 min, it was centrifuged at 3500 r·min -1 for 10 min, and the upper-layer serum was separated from the plasma. The levels of IL-6, IL-1β, and TNF-α in the serum were detected according to the instructions of the ELISA kit.

[0054] 1.8 Statistical analysis

[0055] GraphPad Prism 9.0 software was used to perform statistical analysis on the data of each group. Measurement data were expressed as mean ± standard deviation and one-way ANOVA was used for comparison of data among multiple groups. A P value < 0.05 was considered to indicate a statistically significant difference.

[0056] 2 Results

[0057] 2.1 Effects of SHTB on the neurobehavioral function of CIRI rats

[0058] The results of the Longa score showed that compared with the sham operation group, the neurological deficit score of the model group rats was significantly increased (P < 0.01); compared with the model group, the neurological deficit scores of the low- and high-dose SHTB groups were significantly decreased (P < 0.05, P < 0.01), and the effect of the high-dose SHTB group was more obvious (P < 0.05, P < 0.01), as shown in Table 1.

[0059] Table 1 Effects of SHTB on the neurological deficit score of CIRI rats ( n = 12)

[0060]

[0061] Compared with the sham operation group: *P < 0.05, **P < 0.01; compared with the model group: # P < 0.05, ## P < 0.01. The same applies hereinafter. 2.2 Effect of SHTB on the cerebral infarction area in rats with CIRI

[0062] The results of TTC staining showed that the local right cerebral ischemia and infarction in the model group rats were severe (46.44%); compared with the model group, the cerebral infarction areas in the low- and high-dose SHTB administration groups were significantly reduced (35.64%, 17.76%), and the differences between groups were statistically significant (P < 0.05). See Figure 1 。

[0063] 2.3 Effect of SHTB on the pathological changes of brain tissue in rats with CIRI

[0064] The results of HE staining showed that the nerve cells in the cortical area of the sham operation group had normal morphology, complete structure, clear boundaries, and neat cell arrangement; the cell structure of the model group rats was abnormal, the cell gap was widened, there were many vacuoles around, mostly showing punctate necrosis, and the cell nucleus was significantly shrunk and deeply stained. The cell gap in the SHTB administration group and the positive drug group was slightly wider, the arrangement was slightly disordered, and the phenomena of cell shrinkage and nuclear condensation and deep staining could still be seen, but the degree was significantly alleviated compared with the model group. See Figure 2 。

[0065] 2.4 Effect of SHTB on the levels of inflammatory factors in the serum of rats with CIRI

[0066] The ELISA results showed that compared with the sham operation group, the levels of IL-1β, IL-6, and TNF-α in the serum of the model group rats were significantly increased; compared with the model group, the levels of TNF-α, IL-6, and IL-1β in the serum of the SHTB administration group and the positive drug group rats were significantly decreased. See Figure 3 。

[0067] The above experimental results showed that Shouhui Tongbian Capsule reduced the neurological deficit score of cerebral ischemia-reperfusion injury, promoted the recovery of neurological function, reduced the cerebral infarction area, alleviated the pathological injury of the brain, and reduced the inflammatory level in the serum, and had a significant brain protection effect on cerebral ischemia-reperfusion injury.

[0068] The above embodiments do not impose any formal limitations on the present invention. For those skilled in the art, any modifications, substitutions, etc. made within the scope of the technical essence of the present invention and within the scope of the technical solution of the present invention still fall within the protection scope of the technical solution of the present invention.

Claims

1. Use of Shouhui Tongbian Capsules in the preparation of drugs for preventing or treating ischemic stroke.

2. The use according to claim 1, characterized in that The Shouhui Tongbian capsule is used in preparing a medicine for treating cerebral ischemia-reperfusion injury.

3. The use according to claim 1, characterized in that The Shouhui Tongbian capsule is used in preparing a drug for reducing the neurological deficit score of ischemic stroke and promoting the recovery of neurological function.

4. The use according to claim 1, characterized in that The Shouhui Tongbian capsule is used in preparing a medicine for reducing the area of ​​cerebral infarction caused by ischemic stroke.

5. The use according to claim 1, characterized in that The Shouhui Tongbian capsule is used in preparing a medicine for alleviating pathological brain damage caused by ischemic stroke.

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