Effects Of Electroacupuncture Of Large Intestine Combined With Giant Deficiency On TNF-α And NF-κB In UC Rats

Objective:To comparative observe the expression level of serum tumour necrosis factor-α(TNF-a) and the expression of nuclear factor kappa-B (NF-kB)and nicotinic acetylcholine receptora7(nAChRa7) in colon tissue by using electroacupuncture at five different acupoints-Shangjuxu, Zusanli, Xiajuxu, Yanglingquan and Chengjin on rat ulcerative colitis models, and to exprore whether Shangjuxu has relative specificity when treating its corresponding viscera’s disease.Methods:Seventy SD rats (50%male and50%female) were randomly divided into7groups:a blank group, a model group, Shangjuxu group, Zusanli group, Xiajuxu group, Yanglingquan group and Chengjin group,10rats in a group. The blank group was treated with0.9%NaC1by enama, and the UC rat model was made with by trinitrobenzene sulphoneacid with ethanal enama. Then the five treatment groups respectively received electroaccupuncture at corresponding acupoints for10consecutive days.The electroacupuncture parameters were density wave (10/50HZ),the negative electrode on the right acupoint and the positive on the left, the intensity between1-3mA with limb tremor in degrees, once a day,30min for once.Then colon tissues were collected to make UC score by eye first and histological changes in the colon were observed by light microscopy. ELISA method is used to check serum TNF-a content and the expression of NF-kB in the colon were detected by immunohistochemistry. The expression of nAChRa7in the colon were detected by western blot.Results:1. Histological changes of rat’s colon tissue in each group:①The results of each group’s rats colon UC scores are:compared to the blank group, the rest six groups of colon UC scores were increased with statistically significant difference(P<0.01or P<0.05);compared to the model group, Shangjuxu group and Zusanli group’s were lower with statistically significant difference(P<0.01or P<0.05); compared to Shangjuxu group, the UC scores in Xiajuxu group, Yanglingquan group and Chengjin group increased with statistically significant difference(P<0.01or P<0.05) but Zusanli group’s score had a faint difference without statistically significance (P>0.05); compared to Zusanli group, Xiajuxu and Yanglingquan group’s scores were similar without statistically significance (P>0.05) while Chengjin group’s score was increased with statistically significant difference(P<0.05).②Analyzing rat’s colon tissue’s histological changes under light microscope after HE staining models:the blank group’s colon tissues had no significant pathological changes; ulceration and inflammation of colon can be observed in the model group; the tissue injury changes of Chengjin group was still obvious; pathological lesions of colonic mucosa were reduced in the Shangjuxu group, Zusanli group,Xiajuxu group and Yanglingquan group, and the Shangjuxu group was more obvious.2.The result of serum TNF-α level in each group rats:compared with the blank group, other groups’s TNF-α level increased with statistically significant difference(P<0.01); compared with the model group, the TNF-a content of five treatment groups decreased obviously (P<0.01); compared with Shangjuxu group, TNF-a content in other four treatment groups increased obviously(P<0.01or P<0.05); compared with Zusanli group and Xiajuxu group, Chengjin group’s TNF-αincreased significantly (P<0.05) but Yanglingquan group was on the contrary(P>0.05).3.The result of NF-kB expression in each group rats:compared with the blank group, other groups’s NF-kB expression increased with statistically significant difference(P<0.01); compared with the model group, the NF-kB expression of four groups of lower He-sea points decreased obviously(P<0.01or jP<0.05); compared with Shangjuxu group, NF-kB expression in Xiajuxu group, Yanglingquan group and Chengjin group increased with statistically significant difference(P<0.01or P<0.05) but Zusanli group’s score had a faint difference without statistically significance (P>0.05); compared with Zusanli group and Xiajuxu group, Chengjin group’s NF-kB expression increased significantly (P<0.05) but Yanglingquan group was on the contrary(P>0.05).4.The result of nAChRa7expression in rat colon for each group:Compared with the blank group, the expression of nAChRa7in other groups(except Shangjuxu group) made obvious decrease(P<0.01); compared with the model group, nAChRa7expression in Shangjuxu group,Zusanli group,Xiajuxu group and Yanglingquan group increased signifcantly (P<0.01); compared with Shangjuxu group, the nAChRa7expression of groups including Xiajuxu,Yanglingquan and Chengjin decreased with significant (P<0.01) while Zusanli group had a faint difference without statistically significance (P>0.05); compared with Zusanli group, Xiajuxu group and Yanglingquan group, Chengjin group’s nAChR α7expression decreased significantly (P<0.05).Conclusions:1.Electroacupuncture at Shangjuxu, Zusanli, Xiajuxu, Yanglingquan and Chengjin all could significantly decrease rat’s UC score, serum TNF-a level,colon tissue’s NF-kB expression and increase the expression of colon nAChR a7in UC model rats. It may be achieved by adjusting the factors related to NF-kB signaling pathway, inhibiting the immune response, regulating inflammatory response and improving pathological changes of colonic mucosa.2.The effect of treating UC inflammation in rats when electroacupuncture at Shangjuxu was better than Xiajuxu,Yanglingquan and Chengjin, partially better than Zusanli, which approved that Shangjuxu has some relative specificity when treating UC.3.This experiment’s results partly confirmed that "curing viscera diseases by He points" refers to that the clinical significance of lower He-sea points lies mainly in treating its corresponding viscera’s disease,its specific mechanism still need further study.