Method for screening medicine for treating liver injury caused by polygonum multiflorum based on biochemical indexes and application of method

Abstract

The invention discloses a method for screening a medicine for treating liver injury caused by polygonum multiflorum based on biochemical indexes and application of the method. The method comprises the following steps: constructing a polygonum multiflorum induced liver injury animal model; the target drug is applied to the fleece-flower root-induced liver injury animal model, the fleece-flower root-induced liver injury animal model is administered according to a fixed time period, and the leucoglobulin ratio, the total protein content and the globulin content in the blood plasma of the fleece-flower root-induced liver injury animal model are detected. The leucoglobulin ratio, the total protein content and the globulin content in the rat plasma are used as indexes for evaluating the medicine for treating the liver injury caused by the polygonum multiflorum. According to the scheme, the method for screening the medicine for treating the liver injury of the polygonum multiflorum is simple and easy to implement, the medicine for treating the liver injury of the polygonum multiflorum can be effectively screened by taking the white ball ratio, the total protein content and the globulin content in the blood plasma of an animal with the liver injury caused by the polygonum multiflorum as detection indexes, and a theoretical basis is also provided for clinical prevention and diagnosis of the liver injury of the polygonum multiflorum.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biotechnology, in particular to a method for screening drugs for treating liver injury caused by Radix Polygoni Multiflori based on biochemical indicators and application thereof. BACKGROUND

[0002] Drug-induced liver injury (DILI) refers to the damage to the liver caused by drugs and their metabolites during the treatment of diseases, which is a common adverse drug reaction. Severe cases can lead to liver failure and even death. At present, DILI has become one of the main reasons for drug withdrawal. In recent years, with the increasing application of traditional Chinese medicines (TCMs) worldwide, drug-induced liver injury events caused by TCMs have occurred frequently, which has attracted great attention from the society.

[0003] Radix Polygoni Multiflori is the dried tuber of Polygonum multiflorum Thunb. It is bitter, sweet, astringent and warm in nature, and belongs to the liver, kidney and heart channels. It has multiple tonifying effects such as tonifying liver and kidney, strengthening bones and muscles, and darkening hair and beard. It mainly contains anthraquinones and stilbene glycosides, and is a traditional tonic TCM in China. Research reports show that emodin and rhein, the main anthraquinone components in Radix Polygoni Multiflori, are the main components causing hepatotoxicity.

[0004] According to statistics, the number of cases of liver injury caused by Radix Polygoni Multiflori ranks first among all TCMs. The risk reports of liver injury caused by oral Radix Polygoni Multiflori have been published on the official websites of domestic and foreign drug regulatory agencies, indicating that the problem of liver injury caused by Radix Polygoni Multiflori has attracted high attention at home and abroad. Foreign drug regulatory departments have successively introduced policies for the supervision and even the limitation of Radix Polygoni Multiflori and its preparations. The frequent adverse reactions of TCMs have caused concerns about the safety of TCMs among the public. Clinical and experimental studies have shown that liver injury induced by Radix Polygoni Multiflori is considered to be an immune-mediated idiosyncratic liver injury, but the role of immune response and its mechanism have not been fully elucidated. There is no specific indicator for early diagnosis in clinic, which constitutes a serious obstacle to the internationalization of TCMs. Therefore, it is necessary to find effective early warning signals based on the early stage of liver injury caused by Radix Polygoni Multiflori. SUMMARY

[0005] The present application aims at at least solving one of the technical problems existing in the prior art. To this end, the present application provides a method for screening drugs for treating liver injury caused by Radix Polygoni Multiflori based on biochemical indicators.

[0006] The present application also provides the application of the above method.

[0007] According to a first aspect of the present application, a method for screening drugs for treating liver injury caused by Radix Polygoni Multiflori based on biochemical indicators is provided, which comprises the following steps:

[0008] (1) constructing an animal model of liver injury caused by Radix Polygoni Multiflori;

[0009] (2) using the target drug in the animal model of liver injury caused by Radix Polygoni Multiflori, and administering the drug to the animal model of liver injury caused by Radix Polygoni Multiflori at a fixed time; detecting the ratio of albumin and globulin, the content of globulin, and / or the content of total protein in the plasma of the animal model of liver injury caused by Radix Polygoni Multiflori, and taking the ratio of albumin and globulin, the content of globulin, and / or the content of total protein in the plasma of the rat as an index for evaluating the drug for treating liver injury caused by Radix Polygoni Multiflori.

[0010] In some embodiments of the present application, the ratio of albumin and globulin is the ratio of albumin and globulin.

[0011] In some embodiments of the present application, the establishment of the animal model of liver injury caused by Radix Polygoni Multiflori comprises the following steps: administering rhein to the animal model at a dose of 300-800 mg / kg for 7-28 days.

[0012] In some embodiments of the present application, the animal model comprises a mouse.

[0013] In some embodiments of the present application, the mouse comprises a mouse and a rat.

[0014] In some embodiments of the present application, the dose is 400-600 mg / kg.

[0015] In some embodiments of the present application, the dose is 450-550 mg / kg.

[0016] In some embodiments of the present application, the dose is 500 mg / kg.

[0017] In some embodiments of the present application, the administration time is 7-21 days.

[0018] In some embodiments of the present application, the administration time is 7-15 days.

[0019] In some embodiments of the present application, the administration time is 14 days.

[0020] According to a second aspect of the present application, there is provided an application of the above method, which is an application in the preparation of a drug for treating liver injury caused by Radix Polygoni Multiflori.

[0021] In some embodiments of the present application, the application is an application in the preparation of a system for detecting liver injury caused by Radix Polygoni Multiflori.

[0022] According to some embodiments of the present application, at least the following advantages are achieved: the method for screening the drug for treating the liver injury caused by Radix Polygoni Multiflori is simple and easy to operate, the white globin ratio, globin content and / or total protein content in the plasma of the animal with the liver injury caused by Radix Polygoni Multiflori can be used as the detection index to effectively screen the drug for treating the liver injury caused by Radix Polygoni Multiflori, and a theoretical basis is provided for the clinical prevention and diagnosis of the liver injury caused by Radix Polygoni Multiflori. BRIEF DESCRIPTION OF DRAWINGS

[0023] The present application will be further described below in conjunction with the drawings and examples, in which:

[0024] Figure 1 The figure shows the changes in the total protein and globin content and white globin ratio in the plasma after 14 days of administration of rhein in Example 1 of the present application, in which "****" represents P<0.001. DETAILED DESCRIPTION

[0025] The concept and technical effects of the present application will be described below in conjunction with the examples to fully understand the purpose, features and effects of the present application. Obviously, the described examples are only a part of the examples of the present application, but not all the examples. Based on the examples of the present application, other examples obtained by those skilled in the art without creative labor are within the scope of protection of the present application.

[0026] The test methods used in the examples are conventional methods unless otherwise specified; the materials, reagents and the like used are commercially available unless otherwise specified.

[0027] Example 1: Establishment of the animal model with the liver injury caused by Radix Polygoni Multiflori and screening of the detection index

[0028] In this example, the drug detection model for the liver injury caused by Radix Polygoni Multiflori is constructed, and the detection index for the drug for the liver injury caused by Radix Polygoni Multiflori is screened.

[0029] (1) Establishment of the animal model and preliminary screening of the detection index

[0030] The specific construction method includes the following steps:

[0031] Experimental materials:

[0032] Control drug: sodium carboxymethyl cellulose.

[0033] Solvent preparation (0.5% sodium carboxymethyl cellulose): take 1000 mL ultrapure water in a beaker, heat in a microwave oven, accurately weigh 5.000 g of sodium carboxymethyl cellulose, add to the beaker in small amounts and multiple times, quickly stir to dissolve, and store at 4℃ after visual inspection of uniformity.

[0034] Test drug: rhein (purchased from Aladdin, batch number: R111266).

[0035] Test sample preparation (50 mg / mL rhein): 5 g of rhein was weighed into a mortar, 100 mL of prepared 0.5% sodium carboxymethyl cellulose solution was added, and the mixture was ground until uniform in appearance and then poured into a 250 mL glass bottle.

[0036] SD rats: SPF level 6-8 week old male SD rats were raised in an environment with a temperature of 22-26°C, humidity of 30-70%, 12 h light-dark cycle, and free access to food and water. All experimental animals were acclimated for at least 3 days before the experiment, and the experiment was reviewed by the IACUC committee.

[0037] Experimental method:

[0038] Grouping: 32 SD rats were sorted by weight, and then randomly divided into a control group and a rhein group, 16 rats in each group. The rhein administration group was given 500 mg / kg of rhein per day, and the control group was given the same volume of 0.5% sodium carboxymethyl cellulose per day. The rats were observed and recorded for mental state and feces every day for 14 consecutive days. Blood was collected on the last day for blood biochemical index detection (experimental grouping is shown in Table 1).

[0039] Table 1 Experimental grouping

[0040]

[0041] Experimental results:

[0042] Clinical symptom observation: The rats in the rhein administration group showed loose stool on the third day after administration.

[0043] The blood plasma biochemical level detection results are shown in Figure 1 As can be seen from the figure, the total protein and globulin levels in the plasma of the rats in the administration group were significantly increased, and the albumin and globulin ratio was significantly decreased. Therefore, the total protein content, globulin content, and the ratio of albumin and globulin in the plasma can be used as detection indicators for liver damage caused by Radix Polygoni Multiflori.

[0044] Example 2 A method for screening drugs for treating liver damage caused by Radix Polygoni Multiflori

[0045] The present embodiment provides a method for screening drugs for treating liver damage caused by Radix Polygoni Multiflori, which comprises the following steps:

[0046] (1) Establishing an animal model

[0047] Healthy SD rats were used as administration subjects, and 500 mg / kg of rhein was administered daily for 14 days to construct an animal model of liver damage caused by Radix Polygoni Multiflori;

[0048] (2) Drug screening

[0049] The target drug is used in the animal model of liver injury caused by Radix Polygoni Multiflori, the animal model of liver injury caused by Radix Polygoni Multiflori is dosed at a fixed time, and then the total protein content, globulin content and white ball ratio in the rat plasma are detected, and the total protein content, globulin content and white ball ratio in the rat plasma are used as indexes for evaluating the treatment drug for liver injury caused by Radix Polygoni Multiflori.

[0050] The total protein content, globulin content and white ball ratio in the plasma of the animal model of liver injury caused by Radix Polygoni Multiflori and other detection indexes (such as transaminase) of liver function are also detected, and the results show that the total protein content, globulin content and white ball ratio in the rat plasma can be used as detection indexes of the treatment drug for liver injury caused by Radix Polygoni Multiflori.

[0051] The embodiments of the present application are described in detail above in combination with the drawings, but the present application is not limited to the above embodiments, and various changes can be made within the knowledge range possessed by those skilled in the art without departing from the purpose of the present application. In addition, the embodiments of the present application and the features in the embodiments can be combined with each other without conflict.

Claims

1. A method for screening drugs for treating liver damage caused by Radix Polygoni Multiflori based on biochemical indicators, characterized in that, The method comprises the following steps: (1) constructing a liver injury animal model caused by Radix Polygoni Multiflori; (2) using the target drug in the liver injury animal model caused by Radix Polygoni Multiflori, and administering the liver injury animal model caused by Radix Polygoni Multiflori at a fixed time; detecting the white ball ratio, globulin content and / or total protein content in the plasma of the liver injury animal model caused by Radix Polygoni Multiflori, and taking the white ball ratio, globulin content and / or total protein content in the plasma of the rat as an index for evaluating the liver injury treatment drug of Radix Polygoni Multiflori.

2. The method of claim 1, wherein, The construction of the liver injury animal model caused by Radix Polygoni Multiflori comprises the following steps: administering emodin to the animal model, and the dose is 300-800 mg / kg, and the administration time is 7-28 d.

3. The method of claim 2, wherein, The animal model comprises a mouse.

4. The method of claim 3, wherein, The mouse comprises a mouse and a rat.

5. The method of claim 2, wherein, The dose is 400-600 mg / kg.

6. The method of claim 5, wherein, The dose is 450-550 mg / kg.

7. The method of claim 2, wherein, The administration time is 7-21 d.

8. The method of claim 7, wherein, The administration time is 7-15 d.

9. Use of the method according to any one of claims 1-8 in the preparation of a drug for treating liver injury caused by Radix Polygoni Multiflori.

10. Use of the method according to any one of claims 1-8 in the preparation of a system for detecting liver injury caused by Radix Polygoni Multiflori.

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