阅读说明：本技术 苦参总黄酮在制备治疗溃疡性结肠炎药物中的应用 (Application of total flavonoids of sophora flavescens in preparing medicine for treating ulcerative colitis ) 是由 陈磊 刘怡 邵晶 罗芸 于 2019-11-29 设计创作，主要内容包括：本发明属于医药技术领域,具体涉及苦参总黄酮在制备治疗溃疡性结肠炎药物中的应用。经动物模型实验证明,经口服给药的途径,苦参总黄酮可明显改善溃疡性结肠炎导致的体重下降、结肠组织溃疡、结肠长度变短、结肠出血等情况,并且相较于市面上的皮质类固醇或免疫抑制剂等药物的毒副作用或不良反应小,更加绿色安全。(The invention belongs to the technical field of medicines, and particularly relates to application of sophora flavescens total flavonoids in preparation of a medicine for treating ulcerative colitis. Animal model experiments prove that the lightyellow sophora root total flavonoids can obviously improve the conditions of weight loss, colonic tissue ulcer, colon length shortening, colonic bleeding and the like caused by ulcerative colitis by an oral administration way, and has less toxic and side effects or adverse reactions compared with the drugs such as corticosteroid or immunosuppressant on the market, and is more green and safe.)

1. Application of radix Sophorae Flavescentis total flavonoids in preparing medicine for treating colitis ulcerosa is provided.

2. The use of the total flavonoids of Sophorae radix of claim 1 in the preparation of a medicament for the treatment of ulcerative colitis, wherein the preparation method of the total flavonoids of Sophorae radix comprises the steps of:

s1, adding an ethanol solution into the sophora flavescens, extracting for three times under reflux, extracting for 2 hours each time, and combining the extracting solutions;

s2, removing ethanol in the extracting solution by using a vacuum rotary evaporator, evaporating to dryness in a water bath to obtain an extract, dissolving the extract by using warm water at the temperature of 40-50 ℃, and filtering an aqueous solution of the extract;

s3, extracting the filtered extract water solution with ethyl acetate until the color of an ethyl acetate layer does not change any more, and combining the ethyl acetate layer solution;

s4, recovering ethyl acetate from the ethyl acetate layer solution by a vacuum rotary evaporator to obtain lightyellow sophora root total flavone extract, and freeze-drying to obtain lightyellow sophora root total flavone powder.

3. The use of the kuh-seng total flavonoids of claim 2 in the preparation of a medicament for the treatment of ulcerative colitis, wherein the volume fraction of the ethanol solution in step S1 is 80-95%.

4. The use of the Sophora flavescens ait total flavonoids of claim 2 in the preparation of a medicament for treating ulcerative colitis, wherein the material-to-liquid ratio of Sophora flavescens ait to an ethanol solution in step S1 is 1 (6-9).

5. The application of the sophora flavescens total flavonoids in preparing the medicine for treating ulcerative colitis according to claim 2, wherein the volume ratio of the extract to the warm water in step S2 is 1 (6-10).

6. The use of the Sophora flavescens total flavonoids of claim 2 in the preparation of a medicament for the treatment of ulcerative colitis, wherein the volume ratio of the aqueous extract solution to ethyl acetate in step S3 is 1: 1.

7. The use of the lightyellow sophora root total flavonoids in the preparation of a medicament for treating ulcerative colitis according to claim 1, wherein the medicament is an oral preparation.

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to application of sophora flavescens total flavonoids in preparation of a medicine for treating ulcerative colitis.

Background

Ulcerative Colitis (UC) is a chronic inflammatory bowel disease whose exact mechanism of pathogenesis is not yet clear, and is usually characterized by alternating periods of clinical remission and disease flare-up with the clinical symptoms of abdominal pain, stool dilution, and stool with blood adhesion, weight loss and fever. The pathological manifestations are chronic inflammatory cell infiltration, congestion and erosion of intestinal mucosa. The diseased intestinal segment is primarily the rectum, distal colon, or entire colon. Acute UC has higher mortality, chronic UC is easy to repeat, the recovery possibility is low, and the chronic UC has higher probability to be converted into rectal cancer. In recent years, the incidence rate of UC is on the rise year by year in Asia, and a prospective inflammatory bowel disease population study carried out in 13 countries or regions of Asia-Pacific province shows that the average annual inflammatory bowel disease incidence rate of 13 countries or regions is 1.50/10 ten thousand people, and the annual inflammatory bowel disease incidence rate of China is 3.64/10 ten thousand people, so that it is very important to systematically clarify the pathogenesis of ulcerative colitis and apply proper drug treatment.

The clinical diagnosis and treatment of ulcerative colitis have problems, and firstly, the current diagnosis mainly combines clinical examination, laboratory examination, imaging examination, endoscopy and histopathological representation to carry out comprehensive analysis, and diagnosis is made on the basis of exclusion, and UC and intestinal infection cannot be distinguished; secondly, the existing medicines and therapies have defects, and western medicines for treating UC control acute attack, mucosa repair and maintain and relieve. Aminosalicylic acid and monoclonal antibody drugs are not necessarily effective for all patients, and side effects or adverse reactions of corticosteroids, immunosuppressants and other drugs are excessive. Furthermore, the lack of clinical understanding of the pathogenic heterogeneity in the treatment of UC has led to the failure of the development of several new classes of biological drugs, such as anti-IL 13 drugs for ulcerative colitis.

In traditional Chinese medicine, ulcerative colitis is classified into the categories of "intestinal disease," "chronic dysentery," and "diarrhea" according to clinical symptoms. The chronic dysentery is a more accurate description of the disease due to the persistent and recurrent disease progression of UC. This disease is caused by the affection of damp-heat or food injury or emotional disorder or overstrain, which leads to the impairment of spleen qi, the dampness is endogenous, the damp stagnation is chronic, the heat transformation, the damp-heat fumigation, the stagnation of the intestines, the transmission disorder, the struggle with qi and blood, the blood collaterals are damaged, the qi is coagulated and the blood is stagnated, the blood is spoiled, and the internal ulcer is formed. Damp-heat accumulation in the intestine is always an important factor in the development and progression of UC, and the treatment is based on clearing heat and drying dampness. The mechanism of UC onset is not fully understood in modern medicine. Summarizing the existing research, the currently widely accepted pathogenesis of UC includes mucosal barrier damage, immune barrier abnormalities and biological barrier disorders. The mucosal barrier of the gut includes the tight junctions between the intestinal mucosal epithelium and the intestinal mucus layer. The tight junctions between epithelial cells are composed of proteins such as occludin, junctional adhesion molecules, and occludin. The deficiency of these proteins in the onset of UC results in the failure of fibroblasts to form the typical tight junctions, the breakdown of the intestinal mucosal barrier leading to increased permeability, the translocation of bacteria, endotoxins, etc. to the mucosal lamina propria, and the activation and release of inflammatory factors by immune cells to exacerbate inflammation. In addition, the damage of the intestinal mucosa barrier in UC causes antigenic substances to stimulate the intestinal mucosa and trigger excessive immune activity in the intestinal tract to induce UC. An imbalance of helper T cells and regulatory T cells, and an imbalance of pro-inflammatory and anti-inflammatory cytokines all contribute to UC development. Moreover, the intestinal flora is a biological barrier of the colon mucosa, the intestinal flora of ulcerative colitis patients is obviously different from that of healthy patients, the intestinal microorganism species of UC patients are reduced, the stability of dominant flora is damaged, beneficial bacteria are reduced, pathogenic bacteria are increased, and the secreted endotoxin can damage intestinal epithelial cells and destroy the integrity of the intestinal mucosa.

Evidence of a large number of evidence-based medicines and modern pharmacology proves that the traditional Chinese medicine has obvious advantages and definite curative effect in the aspect of preventing and treating ulcerative colitis and complications thereof, has irreplaceable effect, has the advantage of integrally regulating and controlling multiple systems and multiple targets in traditional Chinese medicine intervention treatment, is wide in clinical application and low in side effect, and is easy to accept by patients. Radix Sophorae Flavescentis is the dry root of Sophora flavescens ait of Sophora of Leguminosae, and has cold and bitter taste, and bitter taste of five flavors, mainly enters heart channel, spleen channel and kidney channel, and has effects of clearing heat and eliminating dampness. It is recorded in Ben Cao gang mu that it has the action of treating intestinal wind and bloody diarrhea and dysentery with heat, and can be used for treating diarrhea, red and white dysentery, abdominal pain, tenesmus, etc. due to gastrointestinal damp-heat. The recent clinical application of radix sophorae flavescentis and compound prescription (such as compound sophora flavescens decoction and fragrant ginseng pills) taking radix sophorae flavescentis as monarch drug for treating various enteritis is also common and has remarkable effect. The main components of the radix sophorae flavescentis are radix sophorae flavescentis alkaloid and kurarinone, a great amount of research on the alkaloid in the radix sophorae flavescentis is carried out by people in the past, and the alkaloid is applied to medicine production, and the research on the kurarinone is relatively late and less. Meanwhile, because the existing lightyellow sophora root is decocted by a traditional water method and is pulverized less (when treating ulcerative colitis, the lightyellow sophora root is pulverized into powder, such as bitter ginseng tablets, fragrant ginseng pills and the like), lightyellow sophora root flavonoid components which are difficult to dissolve in water are discarded along with medicine residues. With the intensive research on the pharmacological actions of sophora flavescens, various pharmacological actions of sophora flavescens flavone such as diabetes resistance, carbohydrate network resistance, anti-inflammation, antibiosis and anticancer are explored, and the extraction and separation of sophora flavescens flavone for separate administration or combined administration with sophora flavescens alkaloid are especially important. In clinical practice, matrine is often used for treating ulcerative colitis, but research on treating ulcerative colitis by matrine is rarely reported.

Summarizing the prior art, the extraction methods of total flavonoids of sophora flavescens can be divided into two main categories. One is alcohol extraction and acid precipitation method, such as patent documents CN200710122642.9, CN201110200778.3, CN201610801529.2, CN201110045288.0, etc., and also such as published documents "structure identification and content determination research of 7 sophora flavescens ait components in sophora flavescens ait total flavone antibacterial extract", "ultrasonic extraction process of sophora flavescens ait total flavone and antibacterial research", etc. Still another is an alcohol extraction organic solvent extraction method, such as patent documents CN200410030938.4, cn200410030937.x, etc., and also documents "extraction separation and biological activity research of kuh-seng flavonoid compound" and "research of flavonoid component in kuh-seng". The alcohol extraction and acid precipitation method needs to firstly extract by ethanol, the extracting solution is acidified by hydrochloric acid and then centrifuged to obtain residue, the residue needs to be washed by water to be neutral and then concentrated and dried to obtain the kuh-seng flavone extract, and the operation of intermediate acidification, centrifugation and water washing is complicated, so that the preparation process of the kuh-seng flavone is needed to be simplified.

Disclosure of Invention

In order to fill the blank of the prior art, the invention provides the application of the lightyellow sophora root total flavonoids in preparing the medicine for treating ulcerative colitis. Animal model experiments prove that the lightyellow sophora root total flavonoids can obviously improve the conditions of weight loss, colonic tissue ulcer, colon length shortening, colonic bleeding and the like caused by ulcerative colitis by an oral administration way, and has less toxic and side effects or adverse reactions compared with the drugs such as corticosteroid or immunosuppressant on the market, and is more green and safe.

Further, the medicament is an oral preparation.

Further, the preparation method of the sophora flavescens total flavonoids comprises the following steps:

s1, adding an ethanol solution into the sophora flavescens, extracting for three times under reflux, extracting for 2 hours each time, and combining the extracting solutions;

s2, removing ethanol in the extracting solution by using a vacuum rotary evaporator, evaporating to dryness in a water bath to obtain an extract, dissolving the extract by using warm water at the temperature of 40-50 ℃, and filtering an aqueous solution of the extract;

s3, extracting the filtered extract water solution with ethyl acetate until the color of an ethyl acetate layer does not change any more, and combining the ethyl acetate layer solution;

s4, recovering ethyl acetate from the ethyl acetate layer solution by a vacuum rotary evaporator to obtain lightyellow sophora root total flavone extract, and freeze-drying to obtain lightyellow sophora root total flavone powder.

Furthermore, the volume fraction of the ethanol solution in the step S1 is 80-95%.

Furthermore, the ratio of the radix Sophorae Flavescentis to the ethanol solution in step S1 is 1 (6-9).

Furthermore, the volume ratio of the extract to the warm water in the step S2 is 1 (6-10).

Furthermore, the volume ratio of the extract water solution to the ethyl acetate in the step S3 is 1: 1.

The preparation method of the sophora flavescens total flavonoids has the advantages of simple steps, easily controlled conditions, suitability for industrial production, high active ingredient content, reliable curative effect and less side effect. Through detection and analysis, the main medicinal components in the sophora flavescens total flavone extract comprise structural compounds such as flavonol, flavanone, chalcone, isoflavone, flavanonol, pterocarpin flavone and the like.

Therefore, compared with the prior art, the invention has the advantages that:

(1) the invention provides application of sophora flavescens total flavonoids in preparing a medicine for treating ulcerative colitis, researches and explains main active ingredients of the sophora flavescens total flavonoids and an action mechanism of the sophora flavescens total flavonoids in treating ulcerative colitis in detail, and fills up the blank of the prior art.

(2) The sophora flavescens total flavonoids extracted by the invention can obviously improve the conditions of weight loss, colonic tissue ulcer, colonic length shortening, colonic bleeding and the like caused by ulcerative colitis, and compared with the drugs such as corticosteroids, immunosuppressants and the like on the market, the sophora flavescens total flavonoids have the advantages of small toxic and side effects or adverse reactions, are more green and safe, and have good market prospect.

Drawings

FIG. 1 is the absorption spectra before and after developing color of norkurarinone control and sample.

FIG. 2 is a graph showing the statistical effect of the weight of rats in each group.

FIG. 3 is a graph showing the statistical effect of groups on the length of rat colon.

FIG. 4 is a colon diagram of rats in each group.

Figure 5 is a graph of the statistics of DAI scores for various groups of rats.

FIG. 6 is a graph showing the results of fecal occult blood tests for various groups of rats.

FIG. 7 is a graph showing the results of HE-stained pathological sections of rats in each group.

Detailed Description

The present invention will be described in detail with reference to specific embodiments, which are illustrative of the invention and are not to be construed as limiting the invention.