CN108904564A - The anti-functional dyspepsia FD active component preparation method and application of cynanchum wilfordii - Google Patents

Abstract

The invention discloses a method for separating active components of Geshanxiao anti-functional dyspepsia and its application. The preparation method is to extract the coarse powder of Geshanxiao medicinal material with 70% ethanol under reflux for three times, and the filtrate is combined to recover ethanol and concentrated until it has no alcohol smell. Adsorbed on ₁₀₁ macroporous resin, eluted with 60% ethanol, recovered ethanol from the eluent and concentrated to extract, and obtained D ₁₀₁ 60% ethanol segment as Geshanxiao anti-functional dyspepsia active component. The present invention screens out that the active component of Geshanxiao against functional dyspepsia is the extract of Geshanxiao D101 60% ethanol segment, thereby providing an application of Geshanxiao in medicine, so that it can be promoted in the field of medicine and help Treat symptoms such as functional dyspepsia.

Description

Preparation method and application of Geshan Xiaokang functional dyspepsia active component

technical field

The invention relates to a preparation method and application of Geshanxiaokang functional dyspepsia active components, belonging to the technical field of medicine.

technical background

Geshanxiao is the lumpy root of Cynanchum auriculatum Royle.et.Wight (Miao medicine name: vob bex teb), also known as Feilaihe, Guoshanxiao, etc. It was first recorded in "Compendium of Materia Medica", and included in the 2003 edition of "Quality Standards for Traditional Chinese Medicine and Ethnic Medicine in Guizhou Province", "Chinese Materia Medica", "Guiyang Folk Medicinal Herbs", etc. It is a commonly used medicinal material for ethnic minorities in Guizhou. It is commonly used by folks to treat digestive system diseases such as anorexia, enteritis, red and white dysentery, and indigestion. Modern pharmacological studies have shown that Geshanxiao has anti-functional dyspepsia, chronic atrophic gastritis, irritable bowel syndrome and other effects.

Functional dyspepsia (FD) is a common disease with frequent occurrence and high recurrence rate, and its pathogenesis is still unclear. According to statistics, the incidence rate of FD population in my country is 18% to 45%, accounting for 20% to 40% of digestive outpatient clinics. With the progress of gastroenterology research in recent years, people have gradually realized that gastrointestinal functional diseases are related to many factors such as changes in visceral sensitivity, changes in inflammatory functions, and changes in the regulatory functions of the central nervous system and enteric nervous system. Studies have found that gastrointestinal dysfunction and gastrointestinal mucosal inflammation are common in FD patients. Professor Jan Tack from the Translational Research Center for Gastrointestinal Disorders in Belgium believes that increased intestinal mucosal permeability may also lead to inflammation and FD. As a commonly used medicinal material for treating indigestion in Guizhou Province, Geshanxiao has a long history of medicinal use. According to domestic and foreign literature reports, Geshanxiao products or total extracts of medicinal materials have anti-depressant, anti-functional dyspepsia, anti-tumor effects, and its chemical components are mainly steroidal glycosides, acetophenones, lignans, Coumarins, organic acids, etc. The current study on the pharmacological activity of Geshan's digestive components, except that 21 steroidal saponins may have antidepressant and antitumor effects, the material basis of other pharmacological activities is still unclear. At present, most of the medicines containing Geshanxiao used clinically are only compound or relatively rough preparations, lacking the development and utilization of active components and active ingredients, lacking objective and scientific quality control indicators of Geshanxiao, and it is difficult to guarantee the clinical application of Geshanxiao related products. curative effect.

Contents of the invention

The purpose of the present invention is to provide a preparation method and application of Geshanxiao anti-functional dyspepsia active components.

Technical scheme of the present invention is as follows:

A method for preparing Geshanxiao anti-functional dyspepsia active components comprises the following steps: take Geshanxiao medicinal material coarse powder and reflux extract it with 70% ethanol for 3 times, combine the filtrate to recover ethanol and concentrate until it has no alcohol smell, and use D ₁₀₁ macroporous resin Adsorbed, eluted with 60% ethanol, recovered ethanol from the eluate and concentrated to the extract to obtain D ₁₀₁ 60% ethanol segment as Geshanxiao anti-functional dyspepsia active component.

Wherein, the aforementioned reflux extraction with 70% ethanol for 3 times includes the following steps: adding 8 times the amount of 70% ethanol for the first time and refluxing for 1.5 hours, adding 6 times the amount of 70% ethanol for the second and third times and refluxing for 1 hour respectively.

The present invention also proposes the application of the active components screened by the above method in the preparation of medicaments for treating functional dyspepsia.

The present invention screens out that the active component of Geshanxiao against functional dyspepsia is the extract of Geshanxiao D10160% ethanol segment, thereby providing an application of Geshanxiao in medicine, so that it can be popularized in the field of medicine and help to treat Symptoms such as functional dyspepsia.

Detailed ways

The present invention will be described in further detail below in conjunction with embodiment.

Example 1:

(1) Preparation of Geshanxiao components

Take 5Kg of Geshanxiao medicinal material coarse powder and extract with 70% ethanol under reflux for 3 times, each time for 3.5 hours, add 8 times the amount of 70% ethanol for the first time and reflux for 1.5 hours, add 6 times the amount of 70% ethanol for the second and third times respectively Reflux for 1 hour, the filtrate was combined to recover ethanol and concentrated to no alcohol smell, adsorbed with D ₁₀₁ macroporous resin, eluted with water, 60% ethanol, and 85% ethanol in sequence, recovered ethanol from the eluate and concentrated to extract, and obtained 4 groups Divide: 70% ethanol extract A, D ₁₀₁ water segment B, D ₁₀₁ 60% ethanol segment C, D ₁₀₁ 85% ethanol segment D.

(2) Screening of active components of Geshanxiao against functional dyspepsia

In the experiment, Geshanxiao components (70% ethanol extract A, D ₁₀₁ water section B, D ₁₀₁ 60% ethanol section C, D ₁₀₁ 85% ethanol section D) were used to study the small intestine propulsion and anti-inflammatory effects.

①Small intestine propulsion test

A: Effects on propulsive function of small intestine in normal mice

Get 60 SPF level Kunming mice, half male and half female, 10 in every group, are randomly divided into 6 groups: blank control group (normal saline), domperidone suspension group (50mg/kg), 70% ethanol extract A , D ₁₀₁ water segment B, D ₁₀₁ 60% ethanol segment C, D ₁₀₁ 85% ethanol segment D. Mice in each group were administered by intragastric administration at 0.2 mL/10 g continuously for 4 days, once a day, and fasted for 12 hours after administration on the third day. After 1 hour of the last administration, 0.2 mL/mouse of 5% charcoal powder suspension (10% gum arabic+5% activated carbon) was administered by intragastric administration respectively. After 30 minutes, the mice were killed by cervical dislocation, and the abdominal cavity was opened to separate the mesentery. Cut the intestinal tube from the upper end to the pylorus and from the lower end to the ileocecal, place it on the table and straighten it, measure the length of the intestinal tube "total length of the small intestine", and use the following formula to calculate the distance from the pylorus to the leading edge of the end of charcoal Charcoal propulsion percentage. (See Table 1).

Charcoal powder propulsion percentage (%) = charcoal powder propulsion distance (cm)/total length of small intestine (cm)×100%

Table 1 The effect of active components of Geshanxiao on the small intestine propulsion function of normal mice ( n=10)

Note: Compared with the blank group, *p<0.05,**p<0.01

B: Effects on small intestinal propulsion dysfunction caused by atropine

Get 70 SPF grade Kunming mice, half male and half male, and are randomly divided into 7 groups, 10 in every group, respectively: blank control group (normal saline), model control group, domperidone suspension group (50mg/kg) And 70% ethanol extract A, D ₁₀₁ water segment B, D ₁₀₁ 60% ethanol segment C, D ₁₀₁ 85% ethanol segment D. Oral administration once a day for 4 consecutive days. Before the last administration, fasting and involuntary water 12h, after administration 30min, except blank control group, all the other 5 groups were intraperitoneally injected atropine sulfate 0.5mg/kg, gavage gave freshly prepared charcoal powder suspension (10% Gum Arabic + 5% activated charcoal) 0.2mL/mouse, the mice were killed by cervical dislocation 20 minutes later, the small intestine was taken out and flattened, and the distance between the full length of the small intestine and the distance from the pyloric sphincter to the end of the small intestine was measured and calculated by the same method as above. (See Table 2).

Table 2 The effect of active components of Geshanxiao on small intestinal propulsion dysfunction caused by atropine ( n=10)

Note: Compared with the blank group, *p<0.05,**p<0.01

② Capillary permeability test

Get 60 KM mice, half male and half female, body weight 18-22g, randomly divided into control group (normal saline), positive drug group, 70% ethanol extract group, D ₁₀₁ water section group, D ₁₀₁ 60% ethanol section, D ₁₀₁ 85% ethanol segment, 10 rats in each group. Oral administration, 2 times a day, continuous administration for 3 days, 1 hour after the last administration in each group, 0.5% Evans blue normal saline solution 10ml/kg was injected into the tail vein, followed by intraperitoneal injection of 0.6% acetic acid 0.2ml/rat. After 20 minutes, the mice were sacrificed, the abdominal wall was cut open, and the peritoneal cavity was washed several times with 6 ml of normal saline, the combined washing fluid was collected, and the volume was adjusted to 10 ml by adding normal saline. Centrifuge at 3000rpm for 15min, take the supernatant, measure the absorbance of the supernatant at 590nm, carry out statistical processing, and obtain the inhibition rate (inhibition rate (%)=(absorbance of blank group-absorbance of drug administration group)/absorbance of blank group× 100%) (see Table 3).

The inhibitory effect of table 3 on the increase of mouse peritoneal capillary permeability caused by acetic acid ( n=10)

Note: Compared with the blank group, *p<0.05,**p<0.01

③ Anti-inflammatory activity study (aspirin-induced gastric mucosal epithelial cell GES-1 injury model)

A: Toxicity of each component of Geshanxiao to GES-1 cells

The experimental results showed that the doses of 70% ethanol extract group, D ₁₀₁ water group, D ₁₀₁ 60% ethanol segment, and D ₁₀₁ 85% ethanol segment had no inhibitory effect on the growth of GES-1 cells when they were not more than 1000 μg/ml (P >0.05), therefore, in the follow-up experiments, the four groups in the following sections all selected the administration concentration not exceeding 1000 μg/ml.

B: Study on the survival rate of aspirin-induced GES-1 damaged cells by each component of Geshanxiao

The results of cell survival rate showed that compared with the model group, Geshanxiao D ₁₀₁ 60% ethanol segment could improve the survival rate of aspirin-injured GES-1 cells to varying degrees, and the dose of 200-400 μg/mL significantly improved the survival rate of injured cells (P<0.05 ) (see Table 4).

The different components of table 4 are to aspirin damage GES-1 cell viability ( n=3)

Note: *P<0.05, **P<0.01vsAspirin group; ^## P<0.01vs Con group

The above is only a preferred example of the present invention, and is not intended to limit any data and form. Any data modification and equivalent changes made to the above embodiments according to the technical essence of the present invention still belong to the technology of the present invention. within a substantial range.

Claims (3)

1. A preparation method for Geshanxiao anti-functional dyspepsia active components, characterized in that: take Geshanxiao medicinal material coarse powder and use 70% ethanol to reflux extract 3 times, and the filtrate is combined and recovered to ethanol and concentrated to no alcohol smell, and is extracted with D ₁₀₁ Adsorbed on macroporous resin, eluted with 60% ethanol, recovered ethanol from eluent and concentrated to extract, and obtained D ₁₀₁ 60% ethanol segment as Geshanxiao anti-functional dyspepsia active component. 2. The preparation method and application of Geshanxiaokang functional dyspepsia active components according to claim 1, characterized in that: the reflux extraction with 70% ethanol for 3 times comprises the following steps: adding 8 times the amount of 70 for the first time % ethanol was refluxed for 1.5 h, and 6 times the amount of 70% ethanol was added for the second and third times respectively, and refluxed for 1 h. 3. Application of the active component screened according to claim 1 or 2 in the preparation of a medicament for treating functional dyspepsia.