Studies On Genes Involved In The Preventive Effect Of Electroacupuncture At Fenglong Acupoint (ST40) On Hypercholesterolemia Mice

The efficiency of electroacupuncture (EA) has been concerned in treating clinical hypercholesterolemia. Our studies have shown that preventive effect of EA at Fenglong acupoint (ST40) on hypercholesterolemia mice. In previous study, the mice models of hypercholesterolemia group (HC), normal group (NM), EA treatment group (EA), simvastatin treatment group (ST), EA preventive group (EP), EA non-acupoint group (EN) were stablished, and the results of blood biochemical indicators manifested that the treatment of EA on fenglong acupoint could markedly prevent the rise of the levels of total cholesterol, high density cholesterol and low density cholesterol of plasma. In order to provide molecular evidence to understand the mechanism of the preventive effect of EA on hypercholesterolemia, identification of differentially expressed genes responding to the hypercholesterolemia have been carried out in HC and EP mice liver.In this study, the livers of HC and EP mice was chosen, the technique of SMART was used to get the total full-length cDNAs from the mice's liver cells from the two groups. Following that, the technique of Suppression Subtractive Hybridization (SSH) was applied in positive and negative directions to acquire the differentially expressed gene profiles. In order to reduce the level of false positives in the two libraries, Negative Subtraction Chain (NSC), a novel subtracted method, was applied to subtracted cDNA populations. Then Dot blot assay was used to select for and amplify differentially expressed cDNAs. Total 22 differential by displayed fragments were cloned and sequenced. The expressions of eleven genes were up-regulated, and five genes were down-regulated in EP mice compared with HC mice. Except two genes whose functions were still unknown, the others were mainly involved in cholesterol metabolism, lipid metabolism, glucose metabolism and immune response.This study was the first time using SSH accompanied with NSC to study gene expression of liver in hypercholesterolemic mice, and optimized the protocol of NSC and SSH. The consequence obtained from this study might be able to arouse our interest to further study the mechanism of the preventive effect of acupuncture on gene level...