Effect Of Shenlingbaizhu Powder On Colonic Epithelial Transport Protein In Rats With Functional Diarrhea And Spleen Deficiency Syndrome

Research BackgroundFunctional diarrhea(FDr)refers to the syndrome of continuous or repeated occurrence of defecation(pastey feces)or watery feces without abdominal pain or abdominal discomfort.The symptoms of diarrhea appeared for at least 6 months,and the symptoms met the diagnostic criteria in the first 3 months,and no gastrointestinal organic changes were found after examination.FDr is a common functional digestive system disease,with intestinal dysfunction as the main clinical manifestation,and belongs to systemic functional disorders.The incidence rate in China is 1.54%,which is at a high level in Asia.It is difficult to cure.It has brought huge economic and medical burdens to patients and society,which seriously affects the quality of life of patients.The pathogenesis of functional diarrhea is not completely clear,but it is mainly related to abnormal gastrointestinal motility,visceral sensory dysfunction,intestinal ion transport disorders,intestinal flora imbalance,gastrointestinal hormones and brain-gut axis abnormalities,social psychological factors,Genetic factors and dietary factors are related.However,there is still insufficient research on the exact pathogenesis,so we still need to dig deeper in this direction.There is no clear classification of functional diarrhea,but it is the same as the classification of diarrhea.According to the duration of the disease,it can be divided into chronic diarrhea and acute diarrhea;according to its pathogenesis,it can be divided into:secretory diarrhea,exudative diarrhea,osmotic diarrhea and gastrointestinal dynamic diarrhea.Although the pathogenesis is not the same,it will eventually lead to imbalance in the absorption and secretion of intestinal water and electrolytes,leading to diarrhea.The imbalance of intestinal water and electrolyte absorption and secretion is achieved through transport proteins,so we will focus on the changes of transport proteins during diarrhea.Because the pathogenesis of diarrhea is not completely clear,modern medicine lacks effective treatments,and most of them are symptomatic treatments.For example,the use of adsorption antidiarrheal drugs to regulate abnormal ion transport,micro-ecological preparations to regulate intestinal flora,intestinal movement inhibitors to slow intestinal peristalsis,and antidepressant and anxiety drugs to improve psychosocial stress in patients.The clinical curative effect is poor and easy to repeat.It cannot fundamentally solve the patient’ s problem,and it is urgent to find new treatment methods.Functional diarrhea belongs to the category of "diarrhea" in Chinese medicine.TCM believes that feelings of external evils,emotional disorders,inadequate diet,chronic illness,and inadequate endowment can all lead to the disease.The site of the disease is in the intestine,and it is also related to the spleen,stomach,liver,and kidneys.The pathological factors are wet,the spleen is weak and wet,but the movement is not healthy.Therefore,the basic principle of traditional Chinese medicine treatment is to strengthen the spleen and dampness,and at the same time take into account the mutual effects of the internal organs,coordinate the internal organs,restore their normal physiological functions,and fundamentally improve the patient’ s symptoms.Shenling Baizhu Powder is a classic representative for the treatment of functional diarrhea with spleen deficiency syndrome.It has the effects of strengthening the spleen,nourishing qi,and dampening and stopping diarrhea.The mechanism of Shenling Baizhu San in improving intestinal ion transport is not clear.This study started with intestinal transporters to study the effect of Shen Ling Bai Zhu San on the abnormal intestinal ion secretion and transport.Research purposesTo observe the effect of Shenling Baizhu Powder on intestinal mucosa and intestinal epithelial cells of functional diarrhea and spleen deficiency model rats,and to explore its mechanism of strengthening spleen and stopping diarrheaResearch contentSixty SPF-grade Wistar rats at 3 weeks of age,(200±5)g.The rats were randomly divided into normal group,model group,positive drug group,Shenlingbaizhu powder high,medium and low dose groups,a total of 6 groups,10 rats in each group.The normal group was fed normally,and the remaining 5 groups were fed with high lactose feed for 14 days.At the same time,the rats were placed on a small platform in a special rectangular water tank from 22:00 to 7:00 the next day,and water was poured around the platform for 14 consecutive days.Method of modeling.During this period,the general condition and body weight,food intake,water intake,grasping power,and fecal water content of the rats were observed.After successful modeling,rats were dosed at 10ml·kg-1,normal group was given normal saline,model group was not given,positive drug group was given montmorillonite powder,and Chinese medicine group was given Shenlingbaizhu powder at different doses.7d.After the treatment was completed,the apical blood and distal colon tissue of each group of rats were measured,and the contents of serum amylase,lactic acid,D-xylose,and 5-HT were measured by colorimetric method or ELISA.The rats were observed by HE staining of pathological tissue sections The pathological changes of the colon were determined by immunohistochemistry and Western blotting methods.Research result1 GeneralAfter modeling,compared with the normal group,the general state of the model group and the remaining 4 groups such as mental state,flexibility,etc.,and the weight,food intake,water consumption,grasping power,etc.were significantly reduced,and the differences were statistically significant(P<0.05),and the diarrhea rate was 100%;compared with the model group,there was no significant difference in the indexes among the other groups of rats(P>0.05).After treatment,compared with the normal group,although the weight of the western medicine group,Chinese medicine high,middle and low dose groups increased to varying degrees,the body weight,food intake,water consumption,and grip strength did not return to normal levels,and the differences were statistically significant.(P<0.05);Compared with the model group,the weight of Western medicine group,Chinese medicine high,medium and low dose groups increased significantly,the difference was statistically significant(P<0.05)The amount of drinking water was significantly increased,and the difference was statistically significant(P<0.05);the grip of the Chinese medicine high and middle dose groups was significantly increased,and the difference was statistically significant(P<0.05).Diarrhea did not reappear in the western medicine group,the Chinese medicine high and medium dose groups,and 12.56%of the rats in the low dose group had diarrhea.Compared with the western medicine group,the body weight and food intake of the Chinese medicine medium-dose group increased significantly,and the differences were statistically significant(P<0.05);the water consumption of the high-and medium-dose groups of Chinese medicine increased significantly,and the differences were statistically significant(P<0.05).2 Serological indicatorsCompared with the normal group,the serum amylase and D-xylose contents of the model group were not significantly different(P>0.05)the serum lactic acid and 5-HT contents of the model group were significantly increased,and the differences were statistically significant(P<0.05).Compared with the model group,the serum amylase content of the rats in the high and low dose groups of Chinese medicine was significantly reduced,and the difference was statistically significant(P<0.05).The serum D-xylose content of the rats in the high,medium and low dose groups of Chinese medicine was significantly increased.The difference was stati stically significant(P<0.05);the levels of serum lactic acid and 5-HT in the western medicine group,Chinese medicine high,medium and low dose groups were significantly reduced,and the difference was statistically significant(P<0.05).Compared with the western medicine group,there were no significant differences in serum amylase,D-xylose,lactic acid,and 5-HT content in the high,middle,and low dose groups of Chinese medicine(P>0.05).3 HE stainingThe conventional pathology of the distal colon showed that the tissue structure of the distal colonic mucosa of the rats in each group was basically normal.The distal colon of the model group,the western medicine group,and the low-dose Chinese medicine group was slightly hyperemic,and no obvious neutrophil infiltration and interval There was no obvious abnormality in the distal colonic tissue of the normal group,the high-dose Chinese medicine group,and the medium-dose group.4 Expression of NHE3,GLUT2,SGLT1,5-HT4 receptor and FFA3 in colon tissue4.1 semi-quantitative results of immunohistochemistryCompared with the normal group,the expression of Na+/H+exchanger 3(NHE3),glucose transporter 2(GLUT2),and sodium-dependent glucose transporter(SGLT1)in the distal colon of the model group was significantly reduced,with differences.Statistical significance(P<0.05);expression of 5-HT4 receptor and free fatty acid receptor 3(FFA3)in the model group was significantly increased,and the differences were statistically significant(P<0.05);Compared with the model group,the expressions of NHE3 and SGLT1 in the western medicine group,high,medium and low dose groups were not significantly different(P>0.05);the expressions of GLUT2 in the high,medium and low dose groups of Chinese medicine were significantly increased,and the differences were statistically significant(P<0.05)the expression of 5-HT4 receptor in the high,medium and low dose groups of Chinese medicine was significantly reduced,and the difference was statistically significant(P<0.05);the expression of FFA3 in the high and medium dose groups of Chinese medicine was significantly reduced,and the difference was statistically significant(P<0.05).Compared with the western medicine group,there were no significant differences in the expressions of NHE3,GLUT2,SGLT1,5-HT4 receptor and FFA3 in the high,medium and low dose groups of Chinese medicine(P>0.05).4.2 Western blotting resultsCompared with the normal group,the expression of NHE3,GLUT2,and SGLT1 in the model group was significantly reduced,and the difference was statistically significant(P<0.05);the expression of 5-HT4 receptor and FFA3 in the model group was significantly increased,and the difference was statistically significant(P<0.05);Compared with the model group,the expressions of NHE3,GLUT2,and SGLT1 in the western medicine group,Chinese medicine high,medium,and low dose groups were significantly increased,and the differences were statistically significant(P<0.05);the western medicine group,Chinese medicine high,medium,and low dose groups 5-HT4 receptor and FFA3 expression were significantly reduced,and the difference was statistically significant(P<0.05);Compared with the western medicine group,the expression of GLUT2 in the high-dose Chinese medicine group was significantly increased,and the difference was statistically significant(P<0.05).Analysis conclusion1 Shenling Baizhu Powder can not only play an antidiarrheal effect,but also improve the hair,mental state,and flexibility of functional diarrhea model rats with spleen deficiency syndrome,promote weight gain,increase the amount of food and water consumed by rats,and increase Grasping power in rats.And in terms of weight gain and food intake,the effect of Shenling Baizhu Powder is better than Montmorillonite Powder.2 Shenling Baizhu Powder can improve the intestinal absorption function of functional diarrhea model rats with spleen deficiency syndrome,increase D-xylose absorption rate,relieve limb soreness,and reduce serum lactic acid concentration.3 Shenling Baizhu Powder can reduce the serum 5-HT concentration in functional diarrhea model rats with spleen deficiency syndrome4 Shenling Baizhu Powder can up-regulate the expression of NHE3,GLUT2,and SGLT1,down-regulate the expression of FFA3 and 5-HT4,and regulate the secretion and absorption of water and electrolytes by the colon,thereby achieving the antidiarrheal effect.