CN109172577B

CN109172577B - Application of burreed tuber component C in rhizoma sparganii in preparation of anti-blood stasis medicine

Info

Publication number: CN109172577B
Application number: CN201810975463.8A
Authority: CN
Inventors: 梁生旺; 冀苏龙; 王淑美; 许晓丽
Original assignee: Guangdong Pharmaceutical University
Current assignee: Guangdong Pharmaceutical University
Priority date: 2018-08-24
Filing date: 2018-08-24
Publication date: 2021-03-16
Anticipated expiration: 2038-08-24
Also published as: CN109172577A

Abstract

The invention disclosesAn application of a scirpusin component C in rhizoma sparganii in the synthesis of a medicament for resisting blood stasis is disclosed. The invention proves that the sparganiin C has obvious effect of resisting blood stasis for the first time, can be used for preparing the medicine for treating blood stasis, and proves that: the bursin C can obviously reduce the thromboxane B in the serum of a blood stasis model mouse₂（Thromboxane B2，TXB₂) The levels of Fibrinogen (FIB), tissue Plasminogen Inhibitor-1 (PAI-1) and Endothelin (endo-1, ET-1) of a body can improve Thymus Index (TI), Spleen Index (SI) and liver index (HI) of mice, and the composition has higher clinical application value and development prospect in the aspect of treatment of blood stasis syndrome.

Description

Application of burreed tuber component C in rhizoma sparganii in preparation of anti-blood stasis medicine

Technical Field

The invention relates to the technical field of medicines, and particularly relates to an application of a sparganium stoloniferum component (sparganium stoloniferum C) in blood stasis resistance.

Background

The rhizoma sparganii is a dry tuber of Sparganium crispum of Sparganium of the family Sparganiaceae, namely Sparganium stolonii Buch. The rhizoma sparganii can be used singly as raw materials or prepared by vinegar, and can also be combined with rhizoma zedoariae and astragalus root to enhance the curative effect, and is mainly used for treating gynecological diseases such as endometriosis and hysteromyoma clinically, and for treating diseases such as liver diseases, cardiovascular and cerebrovascular diseases and the like.

The rhizoma sparganii has complex chemical components and contains flavonoid, volatile oil, phenylpropanoids, organic acid, alkaloid, anthraquinone, steroids, polypeptide and other components. Modern pharmacological studies have elucidated the activities of some compounds and their relationship to clinical use. However, whether a part of the compounds having in vitro activity has a pharmacodynamic action or not requires further research. The method has important significance for guiding clinical medication and development of new drugs.

Disclosure of Invention

The invention aims to solve the technical problem of overcoming the defect and the defect that the effective components of the prior rhizoma sparganii are not clear enough, and the invention is used for the first timeThe effect of the scirpusin A and the scirpusin C on resisting blood stasis is proved. Pharmacological experiments prove that the sparganium A and the sparganium C can obviously reduce TXB in serum of a blood stasis model mouse₂The levels of FIB, PAI-1 and ET-1 can improve the thymus index, the spleen index and the liver index of mice to different degrees, has obvious effect of resisting blood stasis, can be used for preparing a medicament for treating blood stasis, and has higher clinical application value and development prospect.

The invention aims to provide application of a sparganium element component sparganium element C in preparation of a medicine for resisting blood stasis.

The invention also aims to provide a medicament for resisting blood stasis.

The above purpose of the invention is realized by the following technical scheme:

the invention proves that the scirpusin A and the scirpusin C both have obvious blood stasis resisting effect, and have the functions of reducing disease organism Endothelin (endo thelin 1, ET-1), tissue plasminogen Inhibitor-1 (PAI-1) and thromboxane B₂(Thromboxane B2, TXB2) and Fibrinogen (FIB) level, and improves the functions of liver index (HI), Spleen index (Spleen index, SI) and Thymus Index (TI), thereby providing a powerful basis for the development of natural anti-blood stasis drugs.

Therefore, the following applications should be within the scope of the present invention:

application of scirpusin C in synthesizing medicine for resisting blood stasis syndrome is provided.

The application of scirpusin C in preparing a medicine for improving thymus index, spleen index and liver dirty index of patients with blood stasis syndrome is provided.

Preparation of TXB capable of reducing serum of patient with blood stasis syndrome by using sparganin C₂FIB, PAI-1, ET-1 levels.

The burreed rhizome element component is burreed rhizome element C, and the structural formula is as follows:

in addition, the anti-blood stasis medicament containing the scirpusin C also belongs to the protection scope of the invention.

Preferably, the anti-blood stasis medicament also comprises sparganium stoloniferum A.

The anti-blood stasis medicament can also contain a pharmaceutically acceptable carrier and be prepared into different dosage forms.

The invention has the following beneficial effects:

the invention proves that the scirpusin A and the scirpusin C have obvious blood stasis resisting effect for the first time, and experiments prove that the scirpusin C has the effects of reducing Endothelin (endo thelin 1, ET-1) of disease organisms, tissue Plasminogen Inhibitor-1 (Active plasma Activator Inhibitor-1, PAI-1) and thromboxane B₂(Thromboxane B2, TB2) and Fibrinogen (Fibrinogen, FIB) level, and can improve liver index (HI), Spleen index (Spleen index, SI) and Thymus Index (TI), thereby providing a new selection and powerful basis for the development of natural drugs for resisting blood stasis.

Drawings

FIG. 1 shows the effect of sparganium stoloniferum A with different dosages on the comprehensive efficacy index of a mouse organism.

FIG. 2 shows the effect of different dosages of scirpusin B on the comprehensive efficacy index of mouse body.

FIG. 3 shows the effect of different dosages of scirpusin C on the comprehensive efficacy index of mouse body.

Detailed Description

The present invention will be further described with reference to the following specific examples, which are not intended to limit the invention in any way. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.

Unless otherwise indicated, reagents and materials used in the present invention are commercially available.

EXAMPLE 1 functional test of activating blood circulation to dissipate blood stasis

1. Experimental Material

(1) Medicine preparation: the sparganium stoloniferum A, the sparganium stoloniferum B and the sparganium stoloniferum C are prepared by experiments, and the purity is more than or equal to 99%.

(2) Animals: KM mice, female parent, weight 25 + -5 g, provided by Experimental animals center in Guangdong province, license number: SCK 2013-.

(3) Reagent

Epinephrine hydrochloride injection (far-reaching medicine (china) limited, lot number 20170214); bai aspirin enteric-coated tablet (Bayer medicine health promotion Co., Ltd., batch number: BJ 32400); sodium carboxymethylcellulose (CMC-Na, national drug group chemical reagents Co., Ltd., batch No. F20110915, analytical purity); sodium chloride (analytical pure, remote chemical reagents, Inc. of Tianjin, lot number: 20160312); mouse thromboxane B2 (TXB2), endothelin 1(ET-1), plasminogen activator inhibitor 1(PAI-1), Fibrinogen (FIB) enzyme linked immunosorbent assay kit (Tianjin Anorikang Biotechnology Co., Ltd.); the experimental water was pure water.

(4) Laboratory apparatus

A special ultra-pure water machine for WP-UP-WF-10 Watt experiments (Szechwan Watt water treatment equipment Co., Ltd.); ME204E ten thousandth analytical balance (mettler-toledo instruments (shanghai) ltd); one hundred thousand SQP analytical balance (sidoris scientific instruments ltd); YP502N type electronic balance (shanghai precision scientific instruments ltd); LBY-N6K automatic hemorheometer (Beijing Polycosan instruments Co., Ltd.); SIGMA2-16KL low temperature centrifuge (SIGMA); a Thermo microorganism constant temperature incubator (4121); DW-86L626 ultra-low temperature storage box; BIO-RAD iMark enzyme-labeling instrument; IMS-20 snowflake ice maker.

2. Animal grouping:

the mice were randomly divided into a blank group, a model group, an aspirin positive drug control group and a different dosage group of sparganium A according to the body weight, and the administration dosage is shown in the table 1.

TABLE 1 administration of the different dose groups

Note: a represents sparganiin A, B represents sparganiin B, C represents sparganiin C, and the dosage in brackets is multiple of the dosage.

3. Preparing a model:

subcutaneous injection of 1ml kg epinephrine hydrochloride injection in mice^-1After 2 hours, the mixture is put into ice water at the temperature of 0 ℃ for swimming for 3-5 min, taken out and injected again into the body of the patient with the concentration of 1 ml/kg^-1After 2h intervals, the adrenaline hydrochloride injection is placed in ice water at 0 ℃ for swimming for 3-5 min again, and is taken out to form a mouse acute blood stasis model.

4. Sample preparation:

the mice of the blank group and the model group are intragastrically administered with 0.5 percent CMC-Na solution, and the mice of the other administration groups are intragastrically administered with drugs with corresponding concentration, and the dosage is 0.1 ml.10 g^-1. The acute blood stasis model was replicated in each group of mice by gavage for 7 days, 1 time/day, and 2 hours after the administration on the sixth day. After administration for 2h on the seventh day, blood was collected, centrifuged at 3000 Xg for 10min at 4 ℃, and the supernatant was taken out of an EP tube and stored in an ultra-low temperature refrigerator at-80 ℃ for testing.

5. Sample detection

(1) Sample detection:

re-melting the liquid to be detected at the temperature, adopting an enzyme-linked immunosorbent assay, strictly operating according to the detection steps on the kit specification, and respectively detecting TXB₂ET-1, PAI-1, FIB concentrations; after the mice are killed by blood taking, the mice are dissected, the taken liver, spleen and thymus are used for absorbing surface blood stains by physiological saline, filter paper is used for absorbing surface water, an analytical balance is used for weighing, data are recorded, and the organ index of the mice is calculated (table 2); note: mouse organ index is mouse organ weight (mg)/mouse body weight (g).

Carrying out standardization integration treatment on each index, which is shown in a formula 3-1;

V_{normalized value}＝(V_{Administration set}-V_{Model set})/(V_{Normal group}-V_{Model set}) Equation 3-1

The weight coefficient gives opinions according to the frequency of the indexes appearing in the related documents in the last five years and experts, and the weight coefficient of each index is obtained.

149 documents on blood stasis in the last five years are consulted, 47 of which are examined ET-1, 28 are examined PAI-1 and 36 are examined TXB₂And 38, the FIB was examined. According to the frequency of the detected occurrence of each index in the literature and the opinion of disciplinary experts, namely the weight coefficient of ET-1 and FIB is 3, TXB₂And PAI-1 with a weight factor of 2, thymus index, spleen index and liver index of 1. The total blood circulation promoting and stasis removing effect value is a sum of products of the normalized values of the indexes multiplied by the weight coefficient, namely the total blood circulation promoting effect value, and is calculated according to a formula 2.

The indexes are subjected to standardization and integration treatment according to a formula 1 and a formula 2:

V_{normalized value}＝(V_{Administration set}-V_{Model set})/(V_{Normal group}-V_{Model set}) Equation 1

V_TI、V_HI、V_SIRespectively the standardized effect values of the liver index, the thymus index and the spleen index;

V_PAI-1、V_TXB2、V_ET-2、V_FIBrespectively is the standardized effect value of each detected serum index;

V_totalthe combined effect value is obtained.

(2) And (4) evaluating the results:

results show that aspirin positive drugs, sparganium A, sparganium B and sparganium C can obviously reduce the increase of the levels of ET-1, PAI-1, FIB and TXB2 of organisms caused by molding; and meanwhile, HI, TI and SI of the mice can be improved, and the sparganiin A, the sparganiin B and the sparganiin C are proved to have the effect of reducing blood stasis. Compared with an aspirin administration group, the sparganium A has obviously stronger effect of improving TXB2 and FIB concentration in blood stasis model serum; the effect of the sparganium B on improving TXB2 and FIB concentration in blood stasis model serum is obviously stronger, and the effect of reducing TI value increase caused by model building is better than that of an ASP administration group; the effect of scirpusin C on improving the PAI-1 amount and the TI value of the serum index of the model is obviously better than that of ASP, and the effect on reducing the ET-1 concentration of the serum index of the model and improving the SI has little difference between the improvement degrees of the scirpusin C and the ASP. The results are shown in Table 2.

TABLE 2

Note: comparison with model group^*P<0.05，^**p<0.01; comparison with ASP group P<0.05，p<0.01。

The combined effect values of the individual administration groups increased with increasing dose (table 3, fig. 1).

TABLE 3

The invention researches show that the sparganiin A, the sparganiin B and the sparganiin C have the effects of remarkably reducing the levels of ET-1, PAI-1, FIB and TXB2 and improving HI, TI and SI, and the sparganiin A, the sparganiin B and the sparganiin C are proved to have the effect of resisting blood stasis for the first time, so that a powerful basis is provided for the development of natural medicines resisting blood stasis.

The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.

Claims

1. The application of the scirpusin C in preparing the medicine for resisting the blood stasis syndrome is characterized by improving the thymus index, the spleen index and the liver index of a patient with the blood stasis syndrome; reducing TXB in serum of patients with blood stasis₂The FIB, PAI-1 and ET-1 levels, the blood stasis syndrome is acute blood stasis syndrome, and the sparganiin C has the structural formula shown in the specification：