Traditional Chinese and Tibetan medicine composition with effects of relieving arthralgia and removing stasis

Abstract

The invention provides a traditional Chinese and Tibetan medicine composition with the effects of relieving arthralgia and improving venous extravasated blood, and the traditional Chinese and Tibetan medicine composition is specifically characterized in that the cartilage fluorescence intensity of zebra fish is increased, the venous sinus extravasated blood area of zebra fish is reduced, and the cardiac output and the blood flow velocity are improved. Experiments show that the whole effect is very important by using the ligusticum wallichii in multiple amounts in the formula, and the whole effect is very important by using the radix paeoniae rubra, the rhizoma kaempferiae and the notopterygium root in the formula.

Description

Technical Field

[0003] The invention relates to a Chinese and Tibetan medicine composition with the effects of relieving joint pain and removing blood stasis, belonging to the technical field of disinfection and sterilization products. Background Art

[0005] As people's demand for herbal and natural therapies continues to increase, Chinese herbal topical products have become the first choice for more and more people. Compared with chemical drugs, Chinese herbal topical products are more gentle and natural, and will not cause adverse reactions and dependence. This makes Chinese herbal topical products have unique advantages in relieving common problems such as muscle soreness, sprains, fractures, and congestion.

[0006] The development of traditional Chinese medicine (especially Tibetan medicine) in the field of external use has attracted widespread attention and has shown an exciting market prospect in relieving joint pain and removing blood stasis. The herbal plant extracts contained in the common products on the market, such as Chuanxiong, Safflower, Gentiana macrophylla, etc., and precious minerals such as Myrrh and Frankincense, have excellent analgesic and blood stasis removal effects. These ingredients can penetrate the skin, directly hit the source of pain, promote blood circulation and lymphatic drainage, thereby quickly relieving pain and accelerating wound healing. In addition, the unique compatibility between Chinese and Tibetan medicines and the human body also provides a reliable guarantee for the market prospects of Chinese medicine external use products. With people's attention to natural therapies, the market prospects of Chinese medicine external use products will be further expanded. Summary of the invention

[0008] The purpose of the present invention is to provide a Chinese and Tibetan medicine composition with the effects of relieving joint pain and removing blood stasis.

[0009] The technical scheme of the present invention is as follows: a Chinese-Tibetan medicine composition with the effects of relieving joint pain and removing blood stasis, the formula comprising the following components in parts by weight: 20-60 parts of Chuanxiong, 10-30 parts of Corydalis, 10-30 parts of Carthamus, 10-30 parts of Angelica, 10-30 parts of Red Peony Root, 10-30 parts of Frankincense, 6-30 parts of Myrrh, 6-30 parts of Sichuan Pepper, 6-30 parts of Angelica dahurica, 6-30 parts of Notopterygium, 6-30 parts of Solanum Cubeba, 6-30 parts of Artemisia argyi, 6-30 parts of Kaempferia galanga, 6-30 parts of Borneolum, 4 parts of Clematis, and 1 part of Sanguisorba officinalis.

[0010] Preferably, in the above-mentioned Chinese Tibetan medicine composition, the amount of Chuanxiong is twice the amount of Corydalis.

[0011] The present invention is preferably a Chinese-Tibetan medicine composition with the effects of relieving joint pain and removing blood stasis, the formula of which includes the following components in parts by weight: 32 parts of Chuanxiong, 16 parts of Corydalis, 16 parts of Carthamus, 16 parts of Angelica, 12 parts of Red Peony Root, 12 parts of Frankincense, 10 parts of Myrrh, 10 parts of Sichuan Pepper, 10 parts of Angelica dahurica, 8 parts of Notopterygium, 8 parts of Solanum Cubeba, 8 parts of Artemisia argyi, 8 parts of Kaempferia galanga, 7.8 parts of Borneolum, 4 parts of Clematis, and 1 part of Sanguisorba officinalis.

[0012] The present invention also preferably provides a Chinese-Tibetan medicine composition with the effects of relieving joint pain and removing blood stasis, and its preparation method is as follows: for the above-mentioned medicinal materials, borneol and dragon's blood are soaked and dissolved in 20 times the volume of 95% ethanol, and allowed to stand for 24 hours, and the remaining medicinal materials are hot-extracted 3 times with 60% ethanol by volume, the three times are respectively extracted with 4.5 times the weight of the medicinal materials by volume of ethanol for 2 hours, 2.4 times the weight of the medicinal materials by volume of ethanol for 1 hour, and 2.4 times the weight of the medicinal materials by volume of ethanol for 1 hour, the three extracts are combined and allowed to stand for more than 12 hours, the supernatant is taken, the supernatant with borneol and dragon's blood added is combined, and water is added to 1000 parts by volume to obtain.

[0013] In the present invention, the unit of weight volume is mL / g.

[0014] Beneficial effects:

[0015] The Chinese and Tibetan medicine composition provided by the present invention has the effects of relieving joint pain and improving venous congestion, which is specifically manifested in increasing the fluorescence intensity of cartilage, reducing the area of ​​venous sinus congestion, and improving cardiac output and blood flow velocity. The present invention has found through experiments that the presence or absence of a high proportion of Chuanxiong and red peony root, Kaempferia galanga, and Notopterygium in the formula is very important for the overall efficacy. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1-2 Images and data obtained from experiments to relieve joint pain; Figure 3-4 Images and data obtained from the experiment to improve the area of ​​venous sinus congestion in zebrafish; Figure 5 Data graph obtained from an experiment to improve zebrafish cardiac output; Figure 6 To improve the zebrafish blood flow velocity bar graph.

[0018] in Figure 1 This is a typical graph of the fluorescence intensity of zebrafish cartilage after each sample was treated. The white spots in the figure are the fluorescence intensity after treatment; Figure 3 This is a typical diagram of the congestion area of ​​the zebrafish venous sinus after each sample treatment (the dotted box pointed by the double-headed arrow is the congestion area of ​​the zebrafish venous sinus);

[0019] Figure 2 , Figure 4-Figure 6The bar graph of the experimental data of each group of samples, the vertical axis is the number of pixels, the horizontal axis is each sample group, ZC is the normal group, MX is the model group, YX is the positive control group, DB1 is the comparison group 1 (Chuanxiong is halved and red peony root is removed), DB2 is the comparison group 2 (Kaempferia galanga and Notopterygium incisum are removed), and SYZ is the experimental group of the complete formula. In the data graph, compared with the model control group, **p <0.01, ***p <0.001; compared with the experimental group, #p <0.05, ##p <0.01, ###p <0.001. DETAILED DESCRIPTION

[0021] The present invention is further described in detail in conjunction with the following specific examples, and the protection scope of the present invention is not limited to the following examples. The process, conditions, experimental methods, etc. for implementing the invention, except for the contents specifically mentioned below, are common knowledge and common common sense in the field, and the present invention has no special restrictions.

[0022] Example 1. A Chinese Tibetan medicine composition with the effects of relieving joint pain and removing blood stasis (tentative name: Luoyishu) is made from the following raw materials: 32g of Chuanxiong, 16g of Corydalis, 16g of Carthamus, 16g of Angelica, 12g of Red Peony Root, 12g of Frankincense, 10g of Myrrh, 10g of Sichuan Pepper, 10g of Angelica dahurica, 8g of Notopterygium, 8g of Piper cubeba, 8g of Artemisia argyi, 8g of Kaempferia galanga, 7.8g of Borneolum Syntheticum, 4g of Clematidis, and 1g of Sanguisorba officinalis.

[0023] Preparation method: The above-mentioned medicinal materials, borneol and dragon's blood are soaked and dissolved in 160mL of 95% ethanol and allowed to stand for 24 hours. The remaining medicinal materials are hot-extracted with 60% ethanol for 3 times, namely 800mL ethanol extraction for 2 hours, 430mL ethanol extraction for 1 hour, and 430mL ethanol extraction for 1 hour. The three extracts are combined and allowed to stand for more than 12 hours. The supernatant is taken, combined and added with the supernatant of borneol and dragon's blood, and water is added to 1000mL.

[0024] Example 2: A Chinese-Tibetan medicine composition with the effects of relieving joint pain and removing blood stasis is made from the following raw materials (the amount of Chuanxiong is halved, the amount of red peony root is removed, and the amount of safflower is doubled): 16g of Chuanxiong, 16g of Corydalis, 32g of safflower, 16g of Angelica, 12g of frankincense, 10g of myrrh, 10g of Sichuan pepper, 10g of Angelica dahurica, 8g of Notopterygium, 8g of Piper cubeba, 8g of Artemisia argyi, 8g of Kaempferia galanga, 7.8g of borneol, 4g of Clematidis, and 1g of Sanguisorba officinalis.

[0025] The preparation method is the same as Example 1.

[0026] Example 3: A Chinese-Tibetan medicine composition with the effects of relieving joint pain and removing blood stasis is made from the following raw materials (without galangal, notopterygium wilfordii, and double the amount of angelica): 32g of Chuanxiong, 16g of Corydalis yanhusuo, 16g of safflower, 32g of angelica, 12g of red peony root, 12g of frankincense, 10g of myrrh, 10g of Sichuan pepper, 10g of Duhuo, 8g of Artemisia argyi, 7.8g of borneol, 4g of Clematidis, and 1g of Sanguisorba officinalis.

[0027] The preparation method is the same as Example 1.

[0028] Example 4: A Chinese Tibetan medicine composition with the effects of relieving joint pain and removing blood stasis, prepared from the following raw materials: 20g of Chuanxiong, 10g of Corydalis, 10g of Carthamus, 10g of Angelica, 10g of Red Peony Root, 10g of Olibanum, 6g of Myrrh, 6g of Sichuan Pepper, 6g of Angelica dahurica, 6g of Notopterygium, 6g of Piper Cubeba, 6g of Artemisia argyi, 6g of Kaempferia galanga, 6g of Borneolum, 4g of Clematis, and 1g of Sanguisorba officinalis. The preparation method is the same as that of Example 1.

[0029] Example 5: A Chinese Tibetan medicine composition with the effects of relieving joint pain and removing blood stasis, prepared from the following raw materials: 60g of Chuanxiong, 30g of Corydalis, 30g of Carthamus, 30g of Angelica, 30g of Red Peony Root, 30g of Olibanum, 30g of Myrrh, 30g of Sichuan Pepper, 30g of Angelica dahurica, 30g of Notopterygium, 30g of Solanum Cubeba, 30g of Artemisia argyi, 30g of Kaempferia galanga, 30g of Borneolum, 4g of Clematis, and 1g of Sanguisorba officinalis. The preparation method is the same as that of Example 1.

[0030] Example 6: A Chinese Tibetan medicine composition with the effects of relieving joint pain and removing blood stasis, prepared from the following raw materials: 30g of Chuanxiong, 15g of Corydalis, 15g of Carthamus, 15g of Angelica, 15g of Red Peony Root, 15g of Olibanum, 10g of Myrrh, 10g of Sichuan Pepper, 10g of Angelica dahurica, 10g of Notopterygium, 6g of Piper Cubeba, 6g of Artemisia argyi, 6g of Kaempferia galanga, 6g of Borneolum, 4g of Clematis, and 1g of Sanguisorba officinalis. The preparation method is the same as that of Example 1.

[0031] Example 7. A Chinese-Tibetan medicine composition with the effects of relieving joint pain and removing blood stasis is made from the following raw materials: 32g of Chuanxiong, 16g of Corydalis, 16g of Carthamus, 16g of Angelica, 12g of Red Peony Root, 12g of Frankincense, 10g of Myrrh, 10g of Sichuan Pepper, 10g of Angelica dahurica, 8g of Notopterygium, 8g of Solanum Piper, 8g of Artemisia annua, 8g of Kaempferia galanga, 7.8g of Borneolum Syntheticum, 4g of Clematis, and 1g of Sanguisorba officinalis.

[0032] Preparation method: The above-mentioned medicinal materials, borneol and dragon's blood are soaked and dissolved in 160mL of 95% ethanol and left to stand for 24 hours. The remaining medicinal materials are cold-soaked in 10 times the amount of 60% ethanol for 72 hours. The clear liquid is taken and combined with the supernatant of borneol and dragon's blood, and water is added to 1000mL to obtain the product.

[0033] The following experimental examples were commissioned by a third party, Hangzhou Huante Biotechnology Co., Ltd., to conduct experiments to further illustrate the present invention, but do not constitute any limitation to the present invention.

[0034] Experimental Example 1: Effect of relieving joint pain

[0035] 1. Test materials

[0036] Sample preparation information

[0037] Luo Yi Shu (experimental group) was prepared according to Example 1; the two control groups were prepared according to Example 2 and Example 3. The stock solution was taken for use and diluted with standard dilution water as needed.

[0038] Positive control: Chondroitin sulfate A sodium salt (hereinafter referred to as chondroitin sulfate), white powder, batch number I2003201, Shanghai Aladdin Biochemical Technology Co., Ltd., stored at 4℃ away from light. Prepared into 100 mg / mL stock solution with standard dilution water, ready for use.

[0039] Experimental animals

[0040] Zebrafish were kept in aquaculture water at 28°C (water quality: 200 mg instant sea salt was added to 1 L of reverse osmosis water, conductivity was 450-550 µS / cm; pH was 6.5-8.5; hardness was 50-100 mg / L CaCO 3 ), the experimental animal use license number is: SYXK (Zhejiang) 2022-0004. The feeding and management are in accordance with the requirements of the international AAALAC certification (certification number: 001458) IACUC ethics review number: IACUC-2023-5749-01.

[0041] Wild-type AB strain zebrafish and transgenic cartilage green fluorescent zebrafish were bred by natural pair mating. Zebrafish aged 2 days post fertilization (2 dpf) were used for the determination of the maximum detection concentration (MTC) of the sample for relieving joint pain and its efficacy evaluation.

[0042] Instruments, consumables and reagents

[0043] Dissecting microscope (SZX7, OLYMPUS, Japan); CCD camera (VertA1, Shanghai Tusen Vision Technology Co., Ltd., China); electric focus continuous zoom fluorescence microscope (AZ100, Nikon, Japan); precision electronic balance (CP214, OHAUS, USA); fully automatic sample rapid grinding device (JXFSTPRP-24L, Shanghai Jingxin Experimental Equipment Technology Department, China); high-speed refrigerated centrifuge (Heraeus Fresco17, ThermoFisher, Germany); ordinary PCR amplification instrument (T100, BIO-RAD, Singapore); fluorescence quantitative PCR instrument (CFX Connect, BIO-RAD, Singapore); UV-visible spectrophotometer (Nanodrop2000, Thermo, USA); microplate mini centrifuge (BE-6100, Haimen Qilin Bell Instrument Manufacturing Co., Ltd., China); low-skirt 96-well plate (transparent) (HSP9601, Bio-rad, USA); 6-well plate (Zhejiang Bellanbo Biotechnology Co., Ltd., China). Dexamethasone acetate (Batch No. B1828095, Shanghai Aladdin Biochemical Technology Co., Ltd., China); dimethyl sulfoxide (DMSO, Batch No. BCCD8942, Sigma, Switzerland); methylcellulose (Batch No. C2004046, Shanghai Aladdin Biochemical Technology Co., Ltd., China); iTaq Universal SYBR Green Supermix (Cat. No. 1725124, Bio-rad, USA); FastKing cDNA First-Strand Synthesis Kit (Genomic-Free) (Batch No. X1213, Tiangen Biochemical Technology (Beijing) Co., Ltd., China); Universal RNA Extraction TL Kit C (Cat. No. TL2204001643C, Foshan Aowei Biotechnology Co., Ltd., China).

[0044] Detection Methods

[0045] MTC assay

[0046] 2 dpf transgenic cartilage green fluorescent zebrafish were randomly selected in 6-well plates, and 30 zebrafish were treated in each well (experimental group). The samples were given water-soluble (concentrations shown in Table 1), and a normal control group and a model control group were set up at the same time, with a volume of 3 mL per well. Except for the normal control group, the remaining experimental groups were given water-soluble dexamethasone acetate to establish a zebrafish cartilage injury model. After treatment at 28℃ for 3 days, the MTC of the samples on the model zebrafish was measured.

[0047] Evaluation of joint pain relief efficacy (cartilage fluorescence intensity)

[0048] 2 dpf transgenic cartilage green fluorescent zebrafish were randomly selected in 6-well plates, and 30 zebrafish were treated in each well. The samples were given water-soluble (see Table 2), and the positive control chondroitin sulfate was 1000 μg / mL. At the same time, a normal control group and a model control group were set up, and the volume of each well was 3 mL. Except for the normal control group, the other experimental groups were given water-soluble dexamethasone acetate to establish a zebrafish cartilage injury model. After 3 days of treatment at 28℃, 10 zebrafish were randomly selected from each experimental group and photographed under a fluorescence microscope. The data were analyzed and collected using NIS-Elements D3.20 advanced image processing software, and the cartilage fluorescence intensity was analyzed. The statistical analysis results of this indicator were used to evaluate the cartilage injury repair efficacy of the samples. The statistical analysis results were expressed as mean ± SE. SPSS26.0 software was used for statistical analysis, and p < 0.05 indicated that the difference was statistically significant.

[0049] Test results

[0050] MTC

[0051] Under the experimental conditions, the MTC of Luoyishu in relieving joint pain was 0.244 μL / mL. See Table 1 for details.

[0052] Table 1. Results of the experimental study on the concentration of Loyishu in relieving joint pain (n = 30)

[0053]

[0054] Evaluation of joint pain relief efficacy (cartilage fluorescence intensity)

[0055] Under the experimental conditions, the complete formula of Luoyishu has a significant effect on relieving joint pain (the fluorescence intensity of cartilage is significantly enhanced); the effect of relieving joint pain (fluorescence intensity of cartilage) of the incomplete formula of comparison group 1 (reduced by half the amount of Chuanxiong, removed red peony root, doubled the amount of safflower) and comparison group 2 (removed Kaempferia galanga and Notopterygium incisum, doubled the amount of angelica) is significantly reduced. See Table 2 for details. Figure 1 and Figure 2 This indicates that the presence or absence of high-proportion Rhizoma Chuanxiong, Radix Paeoniae Rubra, Kaempferia galanga, and Radix Notopterygii is very important for the overall efficacy.

[0056] Table 2. Experimental results of joint pain relief efficacy evaluation (cartilage fluorescence intensity) of each group of samples (n = 10)

[0057] Group Concentration (μL / mL) Cartilage fluorescence intensity (pixels, mean ± SE) Normal control group (ZC) - 1073633 ± 70271* Model control group (MX) - 812159 ± 63085 Chondroitin sulfate (YX) 1000 μg / mL 1145766 ± 77287** Comparison group 1 (DB1) 0.244 903130 ± 87752# Comparison Group 2 (DB2) 0.244 1076593 ± 44032** Experimental group (SYZ) 0.244 1204252 ± 76623***

[0058] Compared with the model control group, *p < 0.05, **p < 0.01, ***p < 0.001; compared with the experimental group, #p < 0.05, ##p < 0.01

[0059] Example 2: Improving venous congestion

[0060] 1. Test materials

[0061] 1.1. Sample preparation information

[0062] Luo Yi Shu (experimental group) was prepared according to Example 1; the two control groups were prepared according to Example 2 and Example 3. The stock solution was taken for use and diluted with standard dilution water as needed.

[0063] Positive control: Compound Danshen Dropping Pills, brown pills, batch number 190203, Tasly Pharmaceutical Group Co., Ltd., stored in a cool place. Prepared into 10.0 mg / mL stock solution with standard dilution water, used immediately after preparation.

[0064] 1.2. Experimental animals

[0065] Zebrafish were kept in aquaculture water at 28°C (water quality: 200 mg instant sea salt was added to 1 L of reverse osmosis water, conductivity was 450-550 μS / cm; pH was 6.5-8.5; hardness was 50-100 mg / L CaCO 3 ), bred and provided by our company's fish farming center, the experimental animal use license number is: SYXK (Zhejiang) 2022-0004, the breeding and management complies with the requirements of the international AAALAC certification (certification number: 001458), IACUC ethics review number: IACUC-2023-5749-01.

[0066] Wild-type AB strain zebrafish were bred in natural pairs. Zebrafish aged 2 dpf were used for the MTC assay and efficacy evaluation of improving venous congestion.

[0067] 1.3. Instruments, consumables and reagents

[0068] Dissecting microscope (SZX7, OLYMPUS, Japan); CCD camera (VertA1, Shanghai Tusen Vision Technology Co., Ltd., China); precision electronic balance (CP214, OHAUS, USA); 6-well plate (Zhejiang Bellambo Biotechnology Co., Ltd., China); heartbeat blood flow analyzer (ZebraBlood 3.4, ViewPoint Life Sciences, France).

[0069] Methylcellulose (Batch No. C2004046, Shanghai Aladdin Biochemical Technology Co., Ltd., China); Verapamil hydrochloride (Batch No. A2007039, Shanghai Aladdin Biochemical Technology Co., Ltd., China).

[0070] 2. Detection Methods

[0071] 2.1. MTC determination

[0072] 2 dpf wild-type AB strain zebrafish were randomly selected in a 6-well plate, and 30 zebrafish were treated in each well (experimental group). The samples were given water-soluble (concentrations are shown in Table 3), and normal control group and model control group were set up respectively, with a volume of 3 mL per well. After 4 h of sample treatment, except for the normal control group, all other experimental groups were given water-soluble verapamil hydrochloride to establish zebrafish heart failure models. After 1 h of treatment at 28℃, the MTC of the samples on the model zebrafish was measured.

[0073] 2.2. Evaluation of efficacy in improving venous congestion (venous sinus congestion area)

[0074] 2 dpf wild-type AB strain zebrafish were randomly selected in 6-well plates, and 30 zebrafish were treated in each well. The samples of each group were given water solution (see Table 4), the positive control compound Danshen dripping pills were 250 μg / mL, and the normal control group and model control group were set up at the same time, with a volume of 3 mL per well. After 4 h of sample treatment, except for the normal control group, all other experimental groups were given water solution of verapamil hydrochloride to establish zebrafish heart failure model. After 1 h of treatment at 28℃, 10 zebrafish were randomly selected from each experimental group and photographed under a dissecting microscope. The data were collected using NIS-Elements D 3.20 advanced image processing software to analyze the area of ​​zebrafish venous sinus congestion. The statistical analysis results of this indicator were used to evaluate the efficacy of the samples in improving venous congestion. The statistical analysis results were expressed as mean ± SE. SPSS26.0 software was used for statistical analysis, and p < 0.05 indicated that the difference was statistically significant.

[0075] 2.3. Evaluation of efficacy in improving venous congestion (cardiac output and blood flow velocity)

[0076] 2 dpf wild-type AB strain zebrafish were randomly selected in a 6-well plate, and 30 zebrafish were treated in each well (experimental group). The samples of each group were given water solution (see Table 5), the positive control compound Danshen dripping pills were 250 μg / mL, and the normal control group and model control group were set up at the same time, with a volume of 3 mL per well. After 4 h of sample treatment, except for the normal control group, the other experimental groups were given water solution of verapamil hydrochloride to establish the zebrafish heart failure model. After 1 h of treatment at 28℃, 10 zebrafish were randomly selected from each experimental group and placed under the heartbeat blood flow analyzer to record the video. The ViewPoint Application Manager software was used to collect data, analyze the blood flow velocity and cardiac output of zebrafish, and the statistical analysis results of these indicators were used to evaluate the efficacy of the samples in improving venous congestion. The statistical analysis results were expressed as mean ± SE. SPSS26.0 software was used for statistical analysis, and p < 0.05 indicated that the difference was statistically significant.

[0077] 3. Test results

[0078] 3.1. MTC

[0079] Under the experimental conditions, the MTC of Luoyishu in improving venous congestion was 0.244 μL / mL. See Table 3 for details.

[0080] Table 3. Results of the experimental study on the concentration of Loyishu in improving venous congestion (n = 30)

[0081]

[0082] 3.2. Evaluation of efficacy in improving venous congestion (venous sinus congestion area)

[0083] Under the experimental conditions, the complete formula of Luoyishu has a significant effect on improving blood stasis in the meridians (the area of ​​venous sinus congestion is significantly reduced); the effect of improving blood stasis in the meridians (the area of ​​venous sinus congestion) of the incomplete formula of the comparison group 1 (reduced the amount of Chuanxiong by half, removed red peony root, and doubled the amount of safflower) and the comparison group 2 (removed Kaempferia galanga and Notopterygium incisum, and doubled the amount of Angelica sinensis) is significantly reduced. See Table 4 for details. Figure 3 and Figure 4 . This shows that the presence or absence of high-proportion Chuanxiong, red peony root, galangal and notopterygium is very important for the overall efficacy. This shows that the presence or absence of high-proportion Chuanxiong, red peony root, galangal and notopterygium is very important for the overall efficacy.

[0084] Table 4. Results of the evaluation experiment on the efficacy of each group of samples in improving venous congestion (venous sinus congestion area) (n = 10)

[0085] Group Concentration (µL / mL) Sinus congestion area (pixels, mean ± SE) Normal control group (ZC) - 1541 ± 90.8*** Model control group (MX) - 9978 ± 1144 Compound Danshen Dropping Pills (YX) 250 μg / mL 4912 ± 678** Comparison group 1 (DB1) 0.244 6779 ± 369*### Comparison Group 2 (DB2) 0.244 5152 ± 351*** Experimental group (SYZ) 0.244 4448 ± 421***

[0086] Compared with the model control group, *p < 0.05, **p < 0.01, ***p < 0.001; compared with the experimental group, #p < 0.05, ###p < 0.001

[0087] 3.3. Evaluation of efficacy in improving venous congestion (cardiac output and blood flow velocity)

[0088] Under the experimental conditions, the complete formula of Luoyishu has a significant effect on improving blood stasis in the meridians (cardiac output and blood flow velocity); the incomplete formula of the comparison group 1 (reduced the amount of Chuanxiong by half, removed the red peony root, and doubled the amount of safflower) and the comparison group 2 (removed Kaempferia galanga and Notopterygium incisum, and doubled the amount of Angelica sinensis) has a significantly lower effect on improving blood stasis in the meridians (cardiac output and blood flow velocity). See Table 5, Table 6, Figure 5 and Figure 6 This indicates that the presence or absence of high-proportion Rhizoma Chuanxiong, Radix Paeoniae Rubra, Kaempferia galanga, and Radix Notopterygii is very important for the overall efficacy.

[0089] Table 5. Experimental results of evaluation of the efficacy of each sample in improving venous congestion (improving cardiac output) (n = 10)

[0090] Group Concentration (µL / mL) Cardiac output (nL / s, mean ± SE) Normal control group (ZC) - 0.302 ± 0.016*** Model control group (MX) - 0.155 ± 0.022 Compound Danshen Dropping Pills (YX) 250 μg / mL 0.223 ± 0.014* Comparison group 1 (DB1) 0.244 0.220 ± 0.017* Comparison Group 2 (DB2) 0.244 0.196 ± 0.015# Experimental group (SYZ) 0.244 0.261 ± 0.026**

[0091] Compared with the model control group, *p < 0.05, **p < 0.01, ***p < 0.001; compared with the experimental group, #p < 0.05, ##p < 0.01

[0092] Table 6. Experimental results of the efficacy evaluation of each sample in improving venous congestion (improving blood flow velocity) (n = 10)

[0093] Group Concentration (µL / mL) Blood flow velocity (μm / s, mean ± SE) Normal control group (ZC) - 995 ± 43.6*** Model control group (MX) - 523 ± 65.3 Compound Danshen Dropping Pills (YX) 250 μg / mL 755 ± 47.0** Comparison group 1 (DB1) 0.244 679 ± 54.7# Comparison Group 2 (DB2) 0.244 785 ± 70.6* Experimental group (SYZ) 0.244 887 ± 68.8**

[0094] Compared with the model control group, *p < 0.05, **p < 0.01, ***p < 0.001; compared with the experimental group, #p < 0.05, ##p < 0.01.

Claims

1. A Chinese-Tibetan medicine composition with the effects of relieving joint pain and removing blood stasis, characterized in that the formula comprises the following components in parts by weight: 20-60 parts of Chuanxiong, 10-30 parts of Corydalis, 10-30 parts of Carthamus, 10-30 parts of Angelica, 10-30 parts of Red Peony Root, 10-30 parts of Frankincense, 6-30 parts of Myrrh, 6-30 parts of Sichuan Pepper, 6-30 parts of Angelica dahurica, 6-30 parts of Notopterygium, 6-30 parts of Solanum Cubeba, 6-30 parts of Artemisia argyi, 6-30 parts of Kaempferia galanga, 6-30 parts of Borneolum, 4 parts of Clematis, and 1 part of Sanguisorba officinalis.

2. The Chinese-Tibetan medicine composition according to claim 1, characterized in that the amount of Chuanxiong in the formula is twice the amount of Corydalis yanhusuo.

3. The Chinese-Tibetan medicine composition according to claim 1, characterized in that the formula comprises the following components in parts by weight: 32 parts of Chuanxiong, 16 parts of Corydalis, 16 parts of Carthamus, 16 parts of Angelica, 12 parts of Red Peony Root, 12 parts of Frankincense, 10 parts of Myrrh, 10 parts of Sichuan Pepper, 10 parts of Angelica dahurica, 8 parts of Notopterygium, 8 parts of Solanum Cubeba, 8 parts of Artemisia argyi, 8 parts of Kaempferia galanga, 7.8 parts of Borneolum, 4 parts of Clematis, and 1 part of Sanguisorba officinalis.

4. The Chinese-Tibetan medicine composition according to any one of claims 1 to 3, characterized in that it is used to improve joint pain and / or remove blood stasis.

5. The Chinese-Tibetan medicine composition according to claim 4, characterized in that improving joint pain refers to improving articular cartilage, and removing blood stasis specifically refers to reducing the area of ​​venous sinus congestion and improving cardiac output and blood flow velocity.

6. The Chinese-Tibetan medicine composition according to any one of claims 1 to 5, characterized in that the formulated medicinal materials are prepared by cold soaking or hot extraction with 60% ethanol.

7. The Chinese-Tibetan medicine composition according to claim 6 is characterized in that it is prepared by the following method: borneol and dragon's blood are soaked and dissolved in 20 volumes of 95% ethanol and allowed to stand for 24 hours, and the remaining medicinal materials are hot-extracted with 60% ethanol by volume for 3 times, the three times are respectively extracted with 4.5 times the weight of the medicinal materials by volume of ethanol for 2 hours, 2.4 times the weight of the medicinal materials by volume of ethanol for 1 hour, and 2.4 times the weight of the medicinal materials by volume of ethanol for 1 hour, the three extracts are combined and allowed to stand for more than 12 hours, the supernatant is taken, the supernatant with borneol and dragon's blood added is combined, and water is added to 1000 parts by volume to obtain the product.

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