Preliminary Study On Pharmacodynamics And Mechanise Of Dendrobium Huoshanse In Treating Constipation Model Of Spleen Yin Deficiency

Objective:To study the effect of Dendrobium huoshanense(DH)on constipation rats caused by spleen-yin deficiency,to clarify its mechanism of action,and to provide an experimental basis for further research.Method:Part 1:A constipation model of spleen-yin deficiency was established in SD rats.The modeling method is compounded by three factors,and the process is divided into two stages,totaling 30 days.On days 1-14,a combination of excessive fatigue(swimming to endurance limit)and an eating disorder(gastric feeding of cabbage and lard every other day)was used to establish a rat model of spleen-qi deficiency model.The success of the model was evaluated using indicators such as anal temperature,body weight,volume diet,and water consumption to evaluate the model with other indicators;on days 15-23,on the basis of the first stage,add gavage medicines(Evodia,Aconite,Cinnamon)to establish a constipation rat model of spleen-yin deficiency.The success of the model was evaluated using indicators such as the time for the first discharge of black stool,the number of fecal particles,and the moisture content of the feces.After the model is successfully established,the rats were randomly divided into 5 groups: model group,Dendrobium huoshanense high,medium and low dose group,Liu Wei Di Huang Pill group.On the 7th day of administration,record the weight,volume diet,and water consumption of the rat;collect the number of bowel movements in the rat within 12 hours for detect the water content of the feces.Determination of the intestinal carbon advancement rate in the small intestine using the carbon advancement experiment.HE staining was used to observe the pathological changes of colon and ileum in each group.Part 2:The content of SP,VIP in serum,MTL,SS in ileum tissues were detected by ELISA method;RT-q PCR method was used to detect the expression of AQP1-11(Except for AQP10)in the colon tissue of each group of rats;the expression level of AQP1,4,5 in colon tissue was detected by Western blot.Result:Part 1:In the stage of spleen-qi deficiency,compared with the normal group,the weight of the model group rats(P<0.01),anal temperature(P<0.05),volume diet(P<0.01),water consumption(P<0.05)were lower than normal group;At the stage of spleen-yin deficiency,the growth rate of body weight of the model group was lower than that of the normal group(P<0.05),the anal temperature increased(P<0.01),the number of fecal particles at 12 hours(P<0.01),and the fecal water content(P<0.05)were lower than the normal group,and the first black stool was eliminated longer than the normal group(P<0.01).After the administration,compared with the model group,the weight of rats in each group of DH aderminstration groups increased(P<0.01),volume diet increased(P<0.01),the number of defecation particles in 12 hours increased(P<0.01),and the small intestine carbon powder advanced rate increased(P<0.05),the fecal moisture content(P<0.01)increased.The results of HE staining showed that in the ileal tissue,the intestinal mucosal epithelial cells of the normal group were tightly connected,intact and neatly arranged,while the model group had loose,irregularly shaped epithelial cells with inflammatory cell infiltration and immunity.The phenomenon of cell aggregation,and gland atrophy,other drug groups have improved to varying degrees.In the colon tissue,the crypt structure of the normal group was intact,the goblet cells were obvious,and the cells were arranged neatly,while the crypt structure of the model group was changed or even degenerated,the goblet cells were greatly reduced,the inflammatory cells infiltrated,and the basal lymphocytes aggregated.Other administration groups have improved to varying degrees.Part 2:ELISA test results showed that compared with the normal group,the MTL content in the ileal tissue of the model group decreased and the SS content increased(P <0.01);the SP content in the serum decreased and the VIP content increased(P<0.01).Compared with the model group,the MTL content in the ileal tissue of rats in each administration group increased,the SS content decreased(P<0.01),the SP content in serum increased,and the VIP content decreased(P<0.01).RT-q PCR test results showed that compared with the normal group,the m RNA levels of AQP1,AQP6,and AQP9 in the model group were decreased(P<0.05),and the m RNA levels of AQP2,AQP3,AQP4,AQP5,AQP8,and AQP11 were lower Increase(P<0.05).Each dosing group can change the expression of these indicators to varying degrees.Western results showed that compared with the normal group,the protein expression levels of AQP1,4,and 5 were significantly higher than that of the normal group(P<0.05),and the Dendrobium Huoshanense administration group could reduce the expression of AQP4,5 at varying degrees(P<0.05)In the low-dose group,the expression of AQP1 was significantly reduced(P<0.05).Conclusion:Dendrobium huoshanense effectively increased the body weight,volume diet,defecation particles within 12 hours,increased fecal moisture content,improved pathological symptoms of ileum and colon tissue in model rats,and increase the secretion of MTL in the ileal tissue,inhibit the secretion of SS,increase the secretion of SP in the serum of rats,and inhibit the secretion of VIP,promote the expression of AQP1,AQP6,AQP9 m RNA levels in colon tissue,inhibit the expression of AQP2,AQP3,AQP4,AQP5,AQP8,AQP11 m RNA levels,and increase to varying degrees Protein expression levels of AQP1,4,and 5.This indicates that Dendrobium huoshanense has obvious effect on the treatment of spleen-yin deficiency constipation,and its mechanism may be related to regulating the secretion of gastrointestinal hormones and improving the abnormality of body water metabolism.