CN111529554A

CN111529554A - Application of cordyceps sobolifera in preparation of anti-fibrosis drugs

Info

Publication number: CN111529554A
Application number: CN202010254741.8A
Authority: CN
Inventors: 于瑞莲
Original assignee: Nanjing Dinghong Pharmaceutical Technology Co ltd
Current assignee: Nanjing Dinghong Pharmaceutical Technology Co ltd
Priority date: 2020-04-02
Filing date: 2020-04-02
Publication date: 2020-08-14

Abstract

An application of cicada fungus in preparing anti-fibrosis medicine belongs to the field of traditional Chinese medicine and biotechnology. Compared with the existing treatment mode, the traditional Chinese medicine composition has definite disease response, can treat and prevent various fibrosis diseases, can inhibit the secretion of TGF-beta 1 by utilizing cicada fungus, ensures that intestinal interstitial cells do not excessively express adhesion molecules and factors for promoting fibrosis such as VEGF, CTGF and the like, does not adhere ECM and cells, and thereby reverses the fibrosis process; the pharmaceutical effect is good for Crohn's disease in inflammatory bowel diseases, the symptoms such as hematochezia and diarrhea can be obviously relieved, intestinal lesions can be repaired, and fibrosis can be reversed; has good safety and no adverse side effects such as liver and kidney.

Description

Application of cordyceps sobolifera in preparation of anti-fibrosis drugs

Technical Field

The invention belongs to the technical field of traditional Chinese medicines and biology, and particularly relates to application of cordyceps sobolifera in preparation of anti-fibrosis medicines.

Background

Fibrosis is a medical concept, and fibrosis (fibrosis) can occur in various organs, and the main pathological changes are that fibrous connective tissue in organ tissues is increased, parenchymal cells are reduced, and continuous progress can cause structural damage and function reduction of organs and even failure, thereby seriously threatening human health and life.

Worldwide, tissue fibrosis is the leading cause of disability and death in many diseases, and as shown by the statistics in the united states, nearly 45% of patients who are fatal to various diseases can be attributed to tissue fibrosis.

The main reason for tissue fibrosis is the massive increase of TGF-beta 1 content, a key cytokine in the fibrosis process, which is a general switch for causing fibrotic lesions. TGF-. beta.1 is thought to be the major driving force for the progression of various inflammatory and fibrotic diseases. TGF-. beta.1 plays an important role in embryonic development and cell differentiation, regulating cell proliferation, inducing synthetic proteins of the ECM, and regulating immune responses. The TGF-. beta.1/Smad pathway and the ERK-MAPK pathway are thought to be pathways associated with intestinal wall fibrosis.

Fibroblasts secrete Connective Tissue Growth Factor (CTGF) upon activation by TGF-. beta.1. CTGF, as a downstream signaling molecule of the TGF- β 1 pathway, can lead to synthesis of collagen i, iii, fibronectin in the extracellular matrix, wound healing and organ fibrosis that has been observed with overexpression of connective tissue growth factor. Drugs that inhibit CTGF production or action may lead to the development of new therapeutic approaches to control fibrotic diseases in humans.

FN-gamma is a glycoprotein having immunoregulatory, antiviral and antitumor activities, which is produced by stimulating T cells and NK cells with an antigen or mitogen, and is a factor for which many anti-fibrotic effects have been studied at present. Experiments prove that it can inhibit fibroblast activation and collagen gene expression.

Crohn's disease is a typical fibrotic disease of intestinal wall tissue.

Crohn's Disease (CD), a typical condition of inflammatory bowel disease, is an autoimmune disease, manifested by chronic inflammation of the gastrointestinal tract, intestinal fibrosis is one of the characteristic pathological manifestations of Crohn's disease, and repeated chronic transmural inflammatory stimulation of the intestinal tract results in excessive extracellular matrix (ECM) and collagen transitional deposition and scar tissue formation, eventually leading to intestinal luminal narrowing and even ileus. Intestinal inflammation in CD patients is transmural inflammation often accompanied by luminal narrowing and stenosis formation due to excessive extracellular matrix deposition, and these complications are clearly distinct from Ulcerative Colitis (UC). The treatment means aiming at the CD intestinal fibrosis is limited, and the pathological intestinal section is mostly removed through an operation, the fibrosis can not be completely cured and is repeatedly attacked, and the life quality of a patient is seriously influenced.

At present, no medicine capable of reversing tissue fibrosis exists.

Chinese medicinal material Cordyceps cicadae, wild Cordyceps cicadae, is complex of Cordyceps cicadae belonging to Paecilomyces cicadae of Clavicepitaceae and similar Cordyceps sinensis; the infected insect body native to the transected mountain range and Tianmu mountain range is the wild cicada fungus of bamboo cicada, called as cicada fungus or big cicada grass, and mainly produced in Jiangsu and Zhejiang provinces. China has long-term understanding and utilization of cordyceps sobolifera, and the record of processing cordyceps sobolifera is recorded in Lei Gong Pao Zhi treatise of Lei 25989 from north and south, which is 800 years earlier than that of cordyceps sinensis. The "materia medica" (the materia medica of the picture) of Song Chaosu song, the "xi luo" (the width of Song Yao), the "syndrome type materia medica" (the cautious word of Song Dynasty Tang), the "compendium of materia medica" (the compendium of materia medica) of Ming Dynasty Li Zhen and the later pharmacopoeia have recorded effects. After the establishment of new China, the medicinal use of cordyceps sobolifera is recorded in numerous book notes such as Chinese medicine universities, Chinese medicine seas, Chinese medicine dictionaries, Chinese materia medica and the like in numerous materia medica compiled in China or pharmacopoeias. Only two cordyceps sinensis are recorded in the traditional Chinese medicine book, one cordyceps sinensis is cordyceps sinensis, the other cordyceps sinensis is cordyceps sobolifera, the cordyceps sinensis is commonly called as small cordyceps due to large flowers and the cordyceps sobolifera is called as big cordyceps due to small flowers, the cordyceps militaris which is sold in a market more hot at present is cordyceps sinensis, and the cordyceps militaris is not recorded in the traditional Chinese medicine book at all and is only eaten as common food.

In the classification of traditional Chinese medicine properties, cordyceps sobolifera and liquorice; cold; it is non-toxic, enters lung and liver meridians, and is cool in nature. Modern researches show that the cordyceps sobolifera has various pharmacological effects, and can enhance the immunologic function of organisms, regulate the nervous system, relieve fever and pain, improve the renal function, regulate lipid metabolism, resist fatigue, stress, tumors and the like.

The artificial cicada fungus is a fermentation product obtained by fermenting paecilomyces cicadae separated from natural cicada fungus, and comprises mycoplasm (culture medium) and fruiting body, and the components and the effect are similar to those of the natural cicada fungus.

However, no research and report on treating various fibrosis diseases by using natural cordyceps sobolifera, artificial cordyceps sobolifera or compound cordyceps sobolifera exists.

Disclosure of Invention

The technical problem to be solved is as follows: aiming at the technical problems, the invention provides application of cordyceps sobolifera in preparation of anti-fibrosis drugs, so that the problem that no therapeutic drug capable of reversing tissue fibrosis exists at present is solved.

The technical scheme is as follows: application of Cordyceps cicadae in preparing anti-fibrosis medicine is provided.

Preferably, the cicada fungus is natural cicada fungus, artificial cicada fungus, single material or compound material and preparation thereof.

Preferably, the cordyceps sobolifera is complete cordyceps sobolifera, sporocarp of cordyceps sobolifera or culture medium of cordyceps sobolifera.

Preferably, the fibrotic disease is an intestinal wall tissue fibrotic disease.

Preferably, the intestinal wall tissue fibrotic disease is crohn's disease.

Has the advantages that: the cicada fungus is used for treating and/or preventing Crohn's disease and various fibrosis diseases, and compared with the traditional Chinese and western medicines for treating the diseases, the cicada fungus has the following advantages and progressiveness:

(1) the cordyceps sobolifera can be used for treating and/or preventing various fibrosis diseases, and mainly utilizes the fact that cordyceps sobolifera can inhibit secretion of TGF-beta 1, so that intestinal interstitial cells do not excessively express adhesion molecules and factors for promoting fibrosis such as VEGF and CTGF, ECM and cells are not adhered, and the fibrosis process is reversed.

(2) The pharmaceutical effect is good for Crohn's disease in inflammatory bowel diseases, the symptoms such as hematochezia and diarrhea can be obviously relieved, intestinal lesions can be repaired, and fibrosis can be reversed;

(3) has good safety and no adverse side effects such as liver and kidney.

Drawings

FIG. 1 is a 40x comparison of Masson's staining before and after treatment with Cordyceps cicadae;

FIG. 2 is a graph of the reduction of colon damage before and after treatment with Cordyceps cicadae;

FIG. 3 is a diagram of the anti-fibrosis mechanism of cicada fungus;

FIG. 4 is a graph of the therapeutic effect of cicada fungus on Crohn's disease.

Detailed Description

The present invention will be described in detail below with reference to the accompanying drawings and specific embodiments. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out under conventional conditions or conditions recommended by the manufacturers.

Example 1 correlation study of spectrum efficacy of pharmaceutically active substance of Cordyceps cicadae

Taking the protection effect of 10 batches of cordyceps sobolifera on injury caused by IEC-6 (note: DSS-dextran sodium sulfate, IEC-6-mouse intestinal epithelial cells) caused by DSS in different harvesting periods as a reference sequence, and taking the peak area of the HPLC (high performance liquid chromatography) spectrum of the cordyceps sobolifera as a comparison sequence to perform gray level correlation analysis. Because the data have different physical meanings, the data are not necessarily in the same dimension, so that the comparison is inconvenient or a correct conclusion is difficult to obtain during the comparison. Therefore, when performing the gray correlation analysis, it is necessary to perform the non-dimensionalization processing on the data in a unified manner. The dimensionless method used in this study is an averaging method. Using DPS software, p (correlation coefficient) was taken to be 0.5, and calculation was performed.

The results show that: the common peaks (peak numbers) with larger drug effect contribution degree (the association degree is more than 0.8) of the cordyceps sobolifera spore powder to the protection effect of the IEC-6 injury caused by the DSS mainly have the common peaks (peak numbers) with larger drug effect contribution degree (1 is more than 2 is more than 18 is more than 9 is more than 8 is more than 26 is more than 25 is more than 15 is more than 14 is more than 13 is more than 21 is more than 3 is more than 33 is more than 28), and the common peaks (peak numbers) with larger drug effect contribution degree (the association degree is more than 0.8) of the cordyceps sobolifera spore stalks to the protection effect of the IEC-6 injury caused by the DSS mainly have the common peaks (peak numbers) with larger drug effect contribution degree (1 is more than 18 is more than 26 is more than 24 is more than 25 is more than. In conclusion, the protective effect of cordyceps sobolifera on IEC-6 injury caused by DSS has the large drug effect peaks of 1, 2, 8, 9, 13, 14, 15, 21, 24, 25, 26, 28, 29 and 33. It follows that the protective effect of cordyceps sobolifera on DSS-induced damage to intestinal epithelial cells is a result of the co-action of the chemical component groups, and not a result of a single component.

TABLE 1 correlation analysis results of spectral efficiency of Cordyceps sobolifera

Example 2 anti-fibrotic Effect of Cordyceps cicadae

The Crohn's disease model developed mice by a single administration of large doses of TNBS (200 mg/kg). The mice are fasted and not forbidden to be watered one day before the experiment, 60 mice are randomly divided into five groups including a group of blank mice and four groups of TNBS treatment groups, the blank mice are anesthetized after being fasted for 24 hours by 0.14mL of 40% ethanol solution, the TNBS treatment group mice are anesthetized after being fasted for 24 hours by 10% w/v chloral hydrate, the TNBS is dissolved in 40% ethanol solution and is rectally filled into an acute Crohn model with the depth of 3.5-4cm through a 3.5F catheter, and after the enema is finished, the mice are placed at the high head part of the tail part for a few minutes to prevent the liquid medicine from flowing out. After two weeks, randomly collecting colon tissues of a blank group and a TNBS treatment group for Masson staining to observe whether fibers are increased, then randomly dividing four TNBS treatment groups into a model group, a cordyceps sobolifera high dose group (5g/kg), a cordyceps sobolifera medium dose group (2.5g/kg) and a tnBS low dose group (1.25g/kg), not carrying out any treatment on the model group until the experiment is finished, and feeding the other groups with cordyceps sobolifera intragastrically once every day from the third week according to the dose until the fourth week is finished. (Note: unit is-medicine mass mg or g/mouse weight kg)

Collagen fibers were blue and muscle fibers were red by Masson staining of the distal colon (fig. 1). The muscle layer of the blank group was bright red and the thickness of the colon intestinal wall was normal. Compared with the control group, the model group has the advantages that the expression of the collagen fibers of the mucosa, the submucosa and the muscular layer is increased, the gland destruction is obvious, and the intestinal wall is thickened. After cicada fungus treatment, the intestinal wall muscle layer of the high-dose group is bright red, the expression of collagen fibers is obviously reduced, and the intestinal wall thickness is uniform and normal; compared with the model group, the middle-dose group has less collagen fibers and less inflammatory cell infiltration; the intestinal wall of the low-dose group is obviously thickened, the expression of a collagen fiber muscular layer is obviously enhanced, and the mucosal gland is damaged. The score of colon fibrosis (figure 2) shows that cordyceps sobolifera has obvious therapeutic effect on colon fibrosis, and the effect is best in the high-dose group (p < 0.05).

By measuring fibrosis related factors in serum and tissues of each group (figure 3), the model group has obviously increased TGF-beta 1 and CTGF level and obviously reduced IFN-gamma content (P <0.01) compared with a blank group, and the cicada fungus treatment group has obviously reduced TGF-beta 1 and CTGF level (P <0.01) and increased IFN-gamma level (P <0.01) compared with the model group in serum and colon tissues of a high-dosage group; the content of TGF-beta 1 and CTGF in the medium-dose group is reduced, and the obvious difference exists (P <0.05), the content of IFN-gamma in tissues is obviously increased (P <0.05), the content of serum is slightly increased, but the obvious difference does not exist compared with the model group (P > 0.05); the low dose group was statistically insignificant compared to the model group (P > 0.05). Therefore, compared with the model group, the cordyceps sobolifera treatment group can reduce the content of TGF-beta 1 and CTGF in tissues and serum to a certain extent and increase the content of IFN-gamma, wherein the high-dose group has the best effect.

Example 3 therapeutic effects of cicada fungus on Crohn's disease

The structural integrity of small intestinal mucosa under the endoscope is the standard for diagnosing Crohn's disease, and the remission under the endoscope is the clinical evaluation standard. Colon HE staining was used for animal experiments. As shown in fig. 4, the distal colon from histological analysis (HE staining) showed that TNBS caused destruction of crypt structure and reduction of goblet cell number, destruction of epithelial cell layer, infiltration of inflammatory cells, and morphologic normalization of blank colon tissue in mice. In the cicada fungus treatment group, compared with the TNBS group, the colon tissue structure of the mouse is basically complete, and the inflammation symptom is reduced, wherein the improvement is best in the high-dose group. The TNBS model group had an elevated colon histological score compared to the blank group (P <0.01), compared to the cordyceps sobolifera high dose group (P <0.01), and compared to the cordyceps sobolifera medium dose group (P < 0.05). This indicates that high doses of cordyceps sobolifera can repair colon tissue in mice and reduce inflammatory response.

Claims

1. Application of Cordyceps cicadae in preparing anti-fibrosis medicine is provided.

2. The application of cicada fungus in preparing anti-fibrosis drugs according to claim 1, wherein the cicada fungus is a natural cicada fungus, an artificial cicada fungus, a single material or a compound preparation thereof.

3. The application of cicada fungus in preparing anti-fibrosis drugs according to claim 1, wherein the cicada fungus is whole cicada fungus, fruiting body of cicada fungus or culture substrate of cicada fungus.

4. The use of cicada fungus according to claim 1 for preparing an anti-fibrotic drug, wherein the fibrotic disease is a fibrotic disease of intestinal wall tissue.

5. The use of cicada fungus according to claim 4 for preparing an anti-fibrosis drug, wherein the fibrotic disease of intestinal wall tissue is Crohn's disease.