Medicine for treating cerebral hemorrhage in mouse experiment

Abstract

The invention discloses a medicine for treating cerebral hemorrhage in a mouse experiment. The medicine is prepared from the following components: astragalus membranaceus, angelica sinensis, radix paeoniae rubra, ligusticum wallichii, lumbricus, peach kernels, safflower carthamus, rhizoma acori graminei, radix rehmanniae recen, radix ophiopogonis, pinellia ternate, pericarpium citri reticulatae, poria cocos, immature bitter orange, ginseng and oyster. The traditional Chinese medicine composition has the effects of activating and nourishing blood, promoting blood circulation to remove blood stasis, dredging collaterals, dispersing blood stasis, nourishing yin, promoting salivation, suppressing yang, tonifying yin and the like, and can effectively slow down the deterioration of ICH lesion and improve the disease progress. Mouse experiments, collagenase technology and quantitative PCR (polymerase chain reaction) detection prove that the medicine for treating cerebral hemorrhage has a regulation effect on ICH lesion.

Description

Technical Field

[0001] The present invention relates to the technical field of treating cerebral hemorrhage, and in particular to a medicine for treating cerebral hemorrhage in mouse experiments. Background Art

[0002] Intracerebral hemorrhage, a non-traumatic bleeding caused by intraparenchymal vascular rupture, accounts for 20% to 30% of all strokes. It is primarily caused by cerebrovascular disease, specifically hyperlipidemia, diabetes, hypertension, vascular aging, and smoking. Patients with intracerebral hemorrhage often experience sudden increases in blood pressure due to emotional agitation or exertion, which can cause rupture and bleeding of diseased cerebral vessels.

[0003] Intracerebral hemorrhage is an acute cerebrovascular disease with high mortality and disability rates, accounting for 10%-15% of all stroke types. Its pathological mechanisms are complex, and the development of therapeutic drugs faces significant challenges. The mechanisms of action of existing drugs are unclear, and methods for validating these mechanisms are lacking, limiting their potential for the treatment and guidance of ICH. Summary of the Invention

[0004] In order to at least solve or partially solve the above problems, a drug for treating cerebral hemorrhage in mouse experiments is provided, which has the effects of promoting blood circulation and nourishing blood, promoting blood circulation and removing blood stasis, unblocking meridians and dispersing blood stasis, nourishing yin and promoting body fluid, and suppressing yang and replenishing yin. It can effectively slow down the deterioration of ICH lesions and improve the progression of the disease.

[0005] In order to achieve the above object, the present invention provides the following technical solutions:

[0006] The present invention provides a drug for treating cerebral hemorrhage in mice experiments, comprising

[0007] Astragalus, Chinese angelica, red peony root, Chuanxiong, earthworm, peach kernel, safflower, Acorus calamus, raw rehmannia root, Ophiopogon japonicus, Pinellia ternata, tangerine peel, Poria cocos, immature bitter orange, ginseng, and oyster.

[0008] As a preferred technical solution of the present invention, it is composed of the following components:

[0009] Astragalus 30-120g, Angelica 10-20g, Red Peony Root 10-20g, Chuanxiong 20-30g, Earthworm 15-30g, Peach Kernel 10-20g, Carthamus 15-30g, Acorus calamus 15-30g, Rehmannia glutinosa 20-30g, Ophiopogon 1-5g, Pinellia 1-5g, Tangerine Peel 1-5g, Poria 1-5g, Citrus aurantium 1-5g, Ginseng 2-10g, Oyster 2-10g.

[0010] As a preferred technical solution of the present invention, when the medicine is boiled, the ratio of the medicine to water is 1:10.

[0011] As a preferred technical solution of the present invention, when boiling the medicine, it is first soaked in water for 30 minutes.

[0012] As a preferred technical solution of the present invention, the medicine needs to be boiled three times, with the three times being 45 minutes, 30 minutes, and 30 minutes respectively.

[0013] As a preferred technical solution of the present invention, the medicine is boiled and filtered through a 0.45um filter membrane, the filtrate is combined, concentrated under reduced pressure to 250 ml and then refrigerated.

[0014] Compared with the prior art, the present invention has the following beneficial effects:

[0015] The present invention has the effects of promoting blood circulation and nourishing blood, promoting blood circulation and removing blood stasis, unblocking meridians and dispersing blood stasis, nourishing yin and promoting body fluid, and subduing yang and replenishing yin. It can effectively slow the deterioration of ICH lesions and improve disease progression. Experiments using mice, using collagenase technology and quantitative PCR detection, have demonstrated the regulatory effect of the drug for treating cerebral hemorrhage on ICH lesions. DETAILED DESCRIPTION

[0016] The preferred embodiments of the present invention are described below. It should be understood that the preferred embodiments described herein are only used to illustrate and explain the present invention, and are not used to limit the present invention.

[0017] Furthermore, if detailed description of known art is not necessary to illustrate the characteristics of the present invention, it will be omitted.

[0018] Example 1

[0019] The present invention provides a drug for treating cerebral hemorrhage in mice experiments, which is composed of the following ingredients:

[0020] Astragalus (tonifies Qi, 30-120g is commonly used), Chinese Angelica (activates blood circulation and nourishes blood), Red Peony Root, Chuanxiong (activates blood circulation and removes blood stasis), Earthworm, Peach Kernel, Safflower (unblocks meridians and removes blood stasis), Acorus Gramineus, Rehmannia Glutinosa, Ophiopogon (nourishes Yin and promotes fluid production), Pinellia, Tangerine Peel, Poria (tonifies the spleen and removes dampness), Citrus aurantium, Ginseng, Oyster (suppresses Yang and replenishes Yin).

[0021] Furthermore, it is composed of the following ingredients:

[0022] Astragalus 30-120g, Angelica 10-20g, Red Peony Root 10-20g, Chuanxiong 20-30g, Earthworm 15-30g, Peach Kernel 10-20g, Carthamus 15-30g, Acorus calamus 15-30g, Rehmannia glutinosa 20-30g, Ophiopogon 1-5g, Pinellia 1-5g, Tangerine Peel 1-5g, Poria 1-5g, Citrus aurantium 1-5g, Ginseng 2-10g, Oyster 2-10g.

[0023] The drugs were soaked in 10 times the amount of water for 30 minutes, decocted three times (45 minutes, 30 minutes, and 30 minutes), filtered through a 0.45 μm filter membrane, the filtrates were combined, concentrated under reduced pressure to 250 ml, and then refrigerated.

[0024] Specifically, the experimental subjects and groupings were as follows: ICR mice, 3-4 weeks old (21-28 days old), weighing approximately 10-15g. After one week of adaptive feeding, the animals were accurately weighed, numbered, and randomly divided into low-dose Chinese medicine intervention groups A and B, medium-dose Chinese medicine intervention groups A and B, and high-dose Chinese medicine intervention groups A and B, with 6 mice in each group.

[0025] ICH model establishment in the experimental group

[0026] Preparation of ICH model by collagenase injection: fasting for 12 hours and water deprivation for 4 hours before surgery, weighing at room temperature (22±3)°C, and intraperitoneal injection of chloral hydrate (0.03mL / 10g) for anesthesia. Then, fix the mouse in prone position on the stereotaxic instrument, adjust the stereotaxic instrument so that the plane of the incisor groove is 1.0mm lower than the plane of the interauricular line, at which point the anterior and posterior fontanelles are basically in the same plane; trim the hair on the dorsal side of the head, and make a longitudinal incision of about 0.5cm in length after skin disinfection. Expose the skull and bregma: 0.5 mm posterior to the bregma and 2 mm to the right of the midline, drill a circular hole approximately 1.0 mm in diameter with a dental drill, without damaging the dura mater. Insert a microinjector perpendicularly along the drilled hole to a depth of 4.0 mm. Then, slowly and evenly inject 0.1 μl of type VII collagenase (0.2 μl / μL, 0.5 μl) into the brain. The entire injection process should be completed within 2 minutes. Leave the needle in place for 10 minutes after injection. After the microinjector is inserted 0.2 mm, leave the needle in place for an additional 5 minutes. Iodine disinfection and suture the skin incision. Ensure the mouse is breathing spontaneously and keeps warm throughout the surgical procedure. After recovery, maintain a warm and ventilated environment. Modeling of a single mouse takes approximately 20 minutes.

[0027] Dosage

[0028] Preliminary experiments using the median lethal dose (LD50) assay confirmed that the Chinese medicine used was low-toxic or non-toxic. The dosage for each group was calculated based on the equivalent dose of human and animal drugs. Given the dosage per kilogram of body weight of animal A, the conversion formula for the dosage per kilogram of body weight of animal B was calculated: the dosage for animal B (mg) = the dosage for animal A (mg·kg-1) × W (conversion coefficient 6.25) × the body weight of animal B (kg). Continuous gavage began on the first day after successful model establishment and continued for four consecutive weeks. The low-dose group: 5g / (kg·d); the medium-dose group: 10g / (kg·d); and the high-dose group: 20g / (kg·d).

[0029] Detection: The drug was administered by gavage once a day. After 4 weeks, the animals were anesthetized and immediately fixed in a supine position. Blood was collected and placed in a test tube. The blood was centrifuged at 2000 rpm for 15 minutes to obtain drug-containing serum. Fluorescence quantitative PCR and ELISA were used to detect cytokines closely related to cerebral hemorrhage lesions. Fluorescent groups were used to label miRNA to obtain fluorescent probes for hybridization with the chip. Image acquisition and data analysis were performed. The fluorescence intensity of the chip was scanned and the experimental results were converted into digital data for storage. The raw data were analyzed and calculated using the supporting software.

[0030] Results: Collagenase technology and quantitative PCR detection were used to prove the regulatory effect of drugs for treating cerebral hemorrhage on ICH lesions.

[0031] The present invention has the effects of promoting blood circulation and nourishing blood, promoting blood circulation and removing blood stasis, unblocking meridians and dispersing blood stasis, nourishing yin and promoting body fluid, and subduing yang and replenishing yin. It can effectively slow the deterioration of ICH lesions and improve disease progression. Experiments using mice, using collagenase technology and quantitative PCR detection, have demonstrated the regulatory effect of the drug for treating cerebral hemorrhage on ICH lesions.

[0032] Finally, it should be noted that the above descriptions are merely preferred embodiments of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art will be able to modify the technical solutions described in the aforementioned embodiments or substitute equivalents for some of the technical features. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention shall be included within the scope of protection of the present invention.

Claims

1. A drug for treating cerebral hemorrhage in mice experiments, characterized in that: include Astragalus, Chinese angelica, red peony root, Chuanxiong, earthworm, peach kernel, safflower, Acorus calamus, raw rehmannia root, Ophiopogon japonicus, Pinellia ternata, tangerine peel, Poria cocos, immature bitter orange, ginseng, and oyster.

2. The drug for treating cerebral hemorrhage in mice according to claim 1, characterized in that: It is composed of the following ingredients: Astragalus 30-120g, Angelica 10-20g, Red Peony Root 10-20g, Chuanxiong 20-30g, Earthworm 15-30g, Peach Kernel 10-20g, Carthamus 15-30g, Acorus calamus 15-30g, Rehmannia glutinosa 20-30g, Ophiopogon 1-5g, Pinellia 1-5g, Tangerine Peel 1-5g, Poria 1-5g, Citrus aurantium 1-5g, Ginseng 2-10g, Oyster 2-10g.

3. The drug for treating cerebral hemorrhage in mice according to claim 1, characterized in that: When the medicine is boiled, the ratio of the medicine to water is 1:

10.

4. The drug for treating cerebral hemorrhage in mice according to claim 1, characterized in that: When the medicine is boiled, it is first soaked in water for 30 minutes.

5. The drug for treating cerebral hemorrhage in mice according to claim 1, characterized in that: The medicine needs to be boiled three times, with the three times being 45 minutes, 30 minutes, and 30 minutes respectively.

6. The drug for treating cerebral hemorrhage in mice according to claim 1, characterized in that: After the drug is boiled, it is filtered through a 0.45 μm filter membrane, the filtrates are combined, concentrated under reduced pressure to 250 ml and then refrigerated.