The Effects Of The Alcohol Extract Of Humulus Scandens On Ion Channels Of Mouse Intestine

?Objective? In this experiment on the basis of secretory diarrheic mouse model,we treated diarrheic mouses with the alcohol extract of humulus scandens.By detecting the therapeutic effect of the alcohol extract of humulus scandens on diarrheic mouse,and the expression of related genes in the intestinal ion channel,we studied the possible molecular mechanism that how the alcohol extract of humulus scandens cures diarrheic mouse.So these founds provide important theoretical scientific references for the development of new diarrhoea medicines and further use of the resources of humulus scandensa and large scale popularization.?Methods? 1?Chose 30 SPF male mouse(5 ~ 6 weeks,weight of 22 ± 1.02 g),randomly divided into three groups: control group,diarrheic model group,treatment group of humulus scandens(10 in each group).In addition to the control group,we treated the rest of the two groups with senna lavage,1 ml each mouse.After 30 min,we treated the control group and model group mice with 0.6 ml/mouse physiological saline;while treatment group with 0.6 ml/mouse alcohol extract of humulus scandens(flavonoids content 3.47%),gastric 7 days in a row.And each mouse fed in a single cage,bedding filter paper on the bottom of the cage,recording stool property and counting times of mushy stool in the day,observing the mental changes,weight changes,DAI score,thickness of colon intestinal wall and the situation of the small intestine pathological changes,so as to evaluate anti-diarrhoea effect of the alcohol extract of humulus scandens on mouse.2?Executing all mouses in the experimental groups after gavage for 7 days,scissoring and marking the type organization at the bottom of the colon,detecting the gene expression from related ion channel of all the mouses' colon with the RT-PCR: Na ion channels(NHE2,NHE3,NHE8),water ion channels(AQP2,AQP3,AQP4,AQP8)and chloride ion channels(CFTR);detecting the expression of target protein by Western blot,so as to clarify possible molecular mechanism of the alcohol extract of humulus scandens curing diarrhea,respectively in the molecular level and protein level.?Result?1?Compared with model group,in the treatment group the mental recovery,weight changes and recovery of small intestinal villus all show a significant effect;Mouses in the diarrheic group,after gavage 7 days in a row,show listlessness,increase of defecate number,diet and weight loss.And the fecal occult blood test results show that most of the mouse in the three days all detected out a positive fecal occult blood test,of which the model group shows obvious bleeding visible to the naked eyes,and thetreatment group have different degrees of bleeding,which shows that building a successful model.And mouses of the treatment group are healthier than model group,as time goes on,they mainly returned to normal,and fecal occult blood test showed negative;Meanwhile,compared with model group,mouse weight has a rising trend,significant difference(P < 0.05);Compared with model group,DAI score is significantly lower,no significant difference compared with control group(P > 0.05);thickness of intestinal wall,compared with control group,significant difference(P < 0.05),which shows that the treatment effect is remarkable.By histologic observing,small intestinal villus of control group are in alignment,the hierarchical structure is clear,no fluffy interstitial edema,no small angiectasis hyperemia,while disordered arrangement of small intestinal villus in model group,most of the villi falls off,villi on the surface of inflammatory exudation,fluffy interstitial edema,small blood vessels,filling.Treatment group compared with model group symptoms change,small intestinal villus neatly,most inflammatory exudation villi surface disappeared,no fluffy interstitial edema,no small blood vessels,congestion,small intestinal structure and normal tissue is roughly same,which shows that the treatment effect is obvious;2 ?RT-PCR results showed that,compared with the control group,the expression of NHE2,NHE3,AQP3,AQP4,AQP8 of diarrhea group significantly lowered(P < 0.05),of which the expression of NHE8,AQP2 and CFTR no significant change(P > 0.05);Compared with diarrhea group,the expression of treatment group AQP3,AQP4 and AQP8 raised obviously,expression of NHE2,NHE3 no significant change;Western blot results showed that,compared with the blank group,protein expression level of AQP3,AQP4,AQP8 in diarrhea group decreased significantly(P < 0.05);Compared with diarrhea group,protein expression level of AQP3,AQP4,AQP8 in treatment group increased(P < 0.05),with statistical significance.?Conclusion? 1 ? The alcohol extract of humulus scandens showed very good treatment effect on secretory diarrhea mouse,and good protection function of colon intestinal mucosa structure in mouse;2?The alcohol extract of humulus scandens can treat diarrhea,by adjusting the transcription and translation of water channel protein(AQP3,AQP4,AQP8).