The Effect of Electroacupuncture on Intestinal Permeability in Rats with Functional Dyspepsia Based on TLR4/ERK Pathway
Objective:To explore the effect of electroacupuncture on intestinal mucosal permeability in rats with functional dyspepsia(FD)through the TLR4/ERK pathway.Methods:A total of 50 SPF-grade 8-week-old male SD rats were randomly divided into blank group(n=10)and modeling group(n=40).The FD model was established in the modeling group using a combination of Guo's tail clamping stimulation method,irregular diet method and ice physiological saline gavage method.A total of 30 successfully modeled rats were randomly divided into model group,electroacupuncture group and TAK242 group,with 10 rats in each group.The electroacupuncture group received electroacupuncture intervention after needling"Zusanli"(ST 36)and"Taichong"(LR 3),with continuous waves,a frequency of 2 Hz,intensity of 1 mA,30 minutes per session,once daily for 14 days.The TAK242 group was intravenously injected with the TLR4 inhibitor TAK242(0.5 mg/kg)once daily for 14 days.After the intervention,the residual rate in the stomach and the propulsion rate in the small intestine were compared among the groups.The serum concentration of diamine oxidase(DAO)was measured by ELISA.The concentrations of lipopolysaccharide(LPS)and D-lactic acid(D-LA)were measured by biochemical methods.The MBP protein expression was measured by immunohistochemical.The protein expressions of TLR4,MyD88,MEK,ERK,p-MEK and p-ERK1/2 in the duodenal tissue were detected by Western blotting.Results:The gastric residue rate significantly increased in model group compared to the blank group(P＜0.01).After intervention,both the electroacupuncture group and the TAK242 group showed a significant decrease in gastric residue rate compared to the model group(P＜0.01).The electroacupuncture group showed lower gastric residue rate than TAK242 group(P＜0.05).The small intestinal propulsion rate significantly decreased in model group compared to the blank group(P＜0.01).After intervention,both the electroacupuncture group and the TAK242 group showed a significant increase in small intestinal propulsion rate compared to the model group(P＜0.01).The electroacupuncture group showed higher small intestinal propulsion rate than TAK242 group(P＜0.05).Compared to the blank group,the relative expression levels of TLR4 and MyD88 proteins in the duodenal tissue significantly increased in model group(P＜0.01).Compared to the model group,the relative expression levels of TLR4 and MyD88 proteins in the duodenal tissue significantly decreased in the electroacupuncture group and the TAK242 group(P＜0.01).The TAK242 group showed lower relative expression level of TLR4 protein than electroacupuncture group(P＜0.05),while there was no statistical difference in the relative expression level of MyD88 protein between TAK242 group and electroacupuncture group(P＞0.05).Com-pared to the blank group,the protein levels of MEK,ERK,p-MEK and p-ERK1/2 in the duodenal tissue were significantly upregulated in model group(P＜0.01).The electroacupuncture group and TAK242 group showed lower protein levels of MEK,ERK,p-MEK and p-ERK1/2 in the duodenal tissue,than model group(P＜0.01).The electroacupuncture group showed lower protein levels of MEK,ERK,p-MEK and p-ERK1/2 in the duodenal tissue,than TAK242 group(P＜0.05).The model group showed higher average optical density of MBP in the duodenal tissue,than blank group(P＜0.01).The average optical density of MBP in the duodenal tissue was significantly down regulated in electroacupuncture group and TAK242 group compared to the model group(P＜0.01).The electroacupuncture group showed lower average optical density of MBP in the duodenal tissue than TAK242 group(P＜0.05).Compared to the blank group,the levels of serum DAO,D-LA and LPS significantly increased in model group(P＜0.01).After intervention,the levels of serum DAO,D-LA and LPS significantly decreased in electroacupuncture group and TAK242 group(P＜0.01).The electroacupuncture group showed lower levels of serum DAO,D-LA and LPS than TAK242 group(P＜0.05).Conclusion:Electroacupuncture can alleviate the inflammatory response of duodenal tissue in FD rats,and restore intestinal mucosal barrier function,and its mechanism may be related to downregulating the expression of TLR4-ERK pathway related proteins.
万方数据
维普
