CN113633652B

CN113633652B - Application of liquiritin in preparing medicament for treating or preventing enterovirus71 type infection

Info

Publication number: CN113633652B
Application number: CN202111057305.2A
Authority: CN
Inventors: 李晋; 徐尚福; 杜文琪
Original assignee: Zunyi Medical University
Current assignee: Zunyi Medical University
Priority date: 2021-09-09
Filing date: 2021-09-09
Publication date: 2023-06-06
Anticipated expiration: 2041-09-09
Also published as: CN113633652A

Abstract

The application discloses application of glycyrrhizin in preparation of medicines for treating or preventing enterovirus71 type infection and application of medicines for inhibiting enterovirus71 type replication in the technical field of traditional Chinese medicine pharmacy, and researches show that after the application adopts the glycyrrhizin and is administrated by lavage, the expression level of EV71VP1 in brain tissues can be reduced, inflammatory factor IL-6 is reduced, virus replication is inhibited, and further the neurological symptoms of hand-foot-and-mouth disease caused by EV71 are improved.

Description

Application of liquiritin in preparing medicament for treating or preventing enterovirus71 type infection

Technical Field

The invention relates to the technical field of traditional Chinese medicine pharmacy, and relates to application of liquiritin in preparing antiviral drugs, in particular to application of liquiritin in preparing drugs for treating or preventing enterovirus71 type infection.

Background

Enterovirus71 (EV 71) infection has become a major threat to public health worldwide, particularly in infants. Epidemiological studies have shown that EV71 infection is the leading cause of severe and even fatal hand-foot-and-mouth disease. EV71 infection is often accompanied by severe neurological complications such as aseptic meningitis, acute flaccid paralysis, encephalitis, etc., which are considered to be the major disease processes in fatal cases. For central nervous system diseases caused by EV71 infection, no effective antiviral therapeutic drug and vaccine can be applied clinically. Therefore, there is an urgent need to actively develop drugs having anti-EV 71 therapeutic effects and protecting the nervous system.

The existing researches on EV71 resistance of traditional Chinese medicines show that qingkailing, xiyanping, blue-bone oral liquid and other preparations, and single traditional Chinese medicines such as coltsfoot, cordate houttuynia and the like all have EV71 resistance, but the specific antiviral components and mechanisms are still insufficient. However, the pathogenesis of damage to the central nervous system caused by EV71 infection, and the antiviral or neuroprotective mechanisms of traditional Chinese medicines or natural products are ambiguous, and remain bottlenecks that restrict safe and effective treatment. Therefore, the antiviral active ingredients of the traditional Chinese medicine are clear, the action mechanism of the antiviral active ingredients is ascertained, the antiviral active ingredients are one of important ways for preventing and treating EV71 related central nervous system diseases, and a basic research basis is provided for the traditional Chinese medicine treatment of the central nervous system diseases caused by EV71 infection.

Disclosure of Invention

Aiming at the defects of the prior art, the invention provides the application of glycyrrhizin in resisting enterovirus 71.

The invention aims at providing an application of glycyrrhizin or pharmaceutically acceptable salt thereof as an active ingredient in preparing a medicine for treating or preventing enterovirus71 type infection.

The second purpose of the invention is to provide an application of glycyrrhizin or pharmaceutically acceptable salt thereof as an active ingredient in preparing medicines for inhibiting enterovirus71 type replication.

Specifically, the disease caused by enterovirus71 infection comprises one or more of hand-foot-mouth disease, viral angina, aseptic meningitis, brain stem encephalitis, neurogenic pulmonary edema and central nervous system infection.

In particular, the medicament can be prepared into any dosage form, including but not limited to the following: tablets (e.g., coated tablets), capsules, oral liquids, oral granules, oral powders, and injections (e.g., lyophilized powder injection and injectable emulsion).

The glycyrrhizin is commercially available, and researches show that after the glycyrrhizin is used for gastric lavage administration, the EV71VP1 expression level in brain tissues can be reduced, inflammatory factor IL-6 is reduced, virus replication is inhibited, and further hand-foot-mouth disease or other neurological symptoms caused by EV71 are improved, so that the aim of preventing and treating diseases caused by EV71 infection is fulfilled.

Drawings

FIG. 1 is a graph showing the weight change, survival and clinical symptoms of each group in a milk mouse infection experiment;

FIG. 2 is a graph of HE staining results for each group of brain tissue;

FIG. 3 is a graph showing the expression level of EV71VP1 in each group of brain tissues;

FIG. 4 is a graph showing the expression level of inflammatory factor IL-6 in brain tissue of each group.

Detailed Description

The following is a further detailed description of the embodiments:

1. establishing EV71 infected suckling mice model, observing the therapeutic effect of glycyrrhizin on EV71 infected suckling mice

1.1 method of Forming mold

30 ICR milk rats of 3 days old, weight 3+ -0.5 g, and 3 groups of 10 ICR milk rats each, which are (1) blank group (Control group); (2) model group (EV 71 group); (3) glycyrrhizin treatment group (LQ group). The model group and the glycyrrhizin treatment group were subjected to intracranial injection of 50 μl of EV71 virus stock solution to prepare an EV71 infection model, and the blank control group was subjected to intracranial injection of an equivalent amount of physiological saline. On the 2 nd day of molding, the glycyrrhizin treatment group adopts the gastric lavage administration of 0.06mg/g of the glycyrrhizin, and the dosage is 1 time per day for 5 days. The blank and model groups were given the same dose of saline for gastric lavage.

1.2 weight changes, survival and clinical symptom scoring

The body weight of the milk mice was measured, and the change in body weight before and after administration was observed. Clinical symptoms were scored for each group of rats, and the clinical scoring criteria are as follows.

Table 1 is the clinical scoring criteria

1.3 brain histopathological detection

After the 5 th day of administration, the rats were fasted for anesthesia and broken ends, brain tissues were taken and HE stained, and pathological changes of the brain tissues were observed.

1.4 detection of EV71VP1 expression level in brain tissue

After the administration on the 5 th day, the rats are fasted and anesthetized, the brain tissues are taken, RNA is extracted, and the EV71VP1 gene expression quantity of the brain tissues is detected.

1.5 detection of the expression level of inflammatory factor IL-6 in brain tissue

After the 5 th day of administration, the rats were fasted and anesthetized, and the brain tissue was taken, RNA was extracted, and the brain tissue IL-6 expression was examined.

2 experimental results

2.1 weight change, survival and clinical symptom score

After 3-day-old rats were vaccinated, daily weight changes, survival and clinical symptom scores were recorded. Referring specifically to fig. 1, a is a general comparison of model group rats with LQ treated group rats; b is the weight change curve of the rats in each experimental group; c is a survival rate curve of the rats in each experimental group; d is the clinical score for each experimental group. Compared with the blank control group, the weight of the model group of suckling mice is obviously reduced, the survival rate is slightly reduced, and the clinical score is obviously increased. After LQ administration treatment, the weight of the suckling mice is obviously increased, the survival rate is improved, and the clinical score is obviously reduced.

2.2 brain histopathological examination results

As can be seen under the microscope, as shown in fig. 2, a is a blank control group, B is a model group, and C is an LQ treatment group. The brain cortex of the blank control group has a large number of nerve cells and round cells. The nerve cell morphology of the model group is shrunken, and a large amount of inflammatory cell infiltration appears, so that the nerve cell morphology is improved compared with the model group after LQ administration treatment, and the inflammatory cell infiltration is reduced.

2.3 results of detection of EV71VP1 expression level in brain tissue

EV71 family picornaviridae enterovirus (enterovirus) which contains a single-stranded positive-strand RNA genome of about 7kb in length. During viral infection, four capsid proteins (VP 1, VP2, VP3 and VP 4) are translated, with VP1 being dominant in the acquisition and transmission of the virus. As shown in fig. 3, vs blank, #p <0.05; vs model group, P <0.05. In the blank control group, EV71VP1 was not expressed, whereas the model group EV71 VP1mRNA was highly expressed. The EV71 VP1mRNA was significantly reduced in the glycyrrhizin treated group compared to the model group.

2.4 detection results of IL-6 expression level of brain tissue inflammatory factor

IL-6 is a pleiotropic cytokine produced by immune cells and non-immune cells that stimulates the immune response of the body. As shown in fig. 4, vs blank, # P <0.01; vs model group, <0.05; compared with the normal control group, the model group has obviously reduced IL-6mRNA expression, and the IL-6mRNA expression is obviously increased after LQ administration. Indicating that LQ promotes the immunocompetence of the body cells and enhances the resistance of the body cells to EV 71.

The foregoing is merely exemplary embodiments of the present invention, and specific structures and features that are well known in the art are not described in detail herein. It should be noted that modifications and improvements can be made by those skilled in the art without departing from the structure of the present invention, and these should also be considered as the scope of the present invention, which does not affect the effect of the implementation of the present invention and the utility of the patent. The protection scope of the present application shall be subject to the content of the claims, and the description of the specific embodiments and the like in the specification can be used for explaining the content of the claims.

Claims

1. The application of glycyrrhizin or pharmaceutically acceptable salt thereof as active ingredient in preparing medicine for treating or preventing enterovirus71 type infection is provided.

2. The application of glycyrrhizin or pharmaceutically acceptable salt thereof as active ingredient in preparing medicines for inhibiting enterovirus71 type replication is provided.

3. The use according to claim 1, characterized in that: the enterovirus71 type infection can be one or more of hand-foot-mouth disease, viral angina, aseptic meningitis, brain stem encephalitis, neurogenic pulmonary edema and central nervous system infection.

4. The use according to claim 3, wherein the medicament is in the form of a tablet, capsule, oral liquid, oral granule, oral powder or injection.