电针背俞穴对非酒精性脂肪肝病大鼠肝肿瘤坏死因子-α的表达及脂质过氧化的影响

Effect of Electroacupuncture Stimulation of Back-shu Points on Expression of TNF-α and Lipid Peroxidation Reaction in the Liver Tissue in Non-alcoholic Fatty Liver Disease Rats

目的:探讨肿瘤坏死因子-α(TNF-α)在非酒精性脂肪肝病(NAFLD)发病机制中的作用并观察电针背俞穴的疗效及作用途径。方法:将Wistar大鼠随机分为正常组(11只)和造模组(32只),造模组饲以高脂饮食复制NAFLD大鼠模型,8周后随机处死正常组1只和造模组2只,病理组织学检查以验证造模成功。然后将余下的造模组大鼠随机分为3组:NAFLD模型组(10只)、电针组(10只)、药物组(10只)。药物组将1%的东宝肝泰混悬液按照0.28g/kg(20mL/kg)灌胃,1次/d;电针组选取双侧背俞穴"脾俞""肾俞""膈俞"进行电针治疗,1次/d,每次20min。连续治疗4周后处死所有动物,HE染色观察肝组织病理学变化情况;全自动生化分析仪测定肝内游离脂肪酸(FFA)含量,比色法检测肝内丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性;免疫组化法检测肝组织中TNF-α的表达。结果:与正常组比较,模型组大鼠均出现中度至重度的肝细胞脂肪变性,肝组织TNF-α表达上调,FFA、MDA升高(P<0.01),SOD活性降低(P<0.01);与模型组比较,电针组和药物组肝组织脂肪变性得到不同程度的减轻,肝组织TNF-α表达下调,FFA、MDA均有不同程度的降低(P<0.01,P<0.05),SOD活性升高(P<0.01,P<0.05)。结论:电针背俞穴可抑制非酒精性脂肪肝大鼠TNF-α表达的上调,降低FFA的释放和减轻脂质过氧化反应,这可能是其治疗NAFLD的作用机制之一。

Objective To observe the effect of electroacupuncture （EA） stimulation （EAS） of back-shu acupoints on expression of tumor necrosis factor-α（（TNF-α） and lipid peroxidase reaction in the liver in non-alcoholic fatty liver disease （NAFLD） rats. Methods Wistar rats were randomly divided into normal group （ n = 11 ）, model group （n = 10）, EAS group （ n = 10） and medication group （n = 10）. The NAFLD model was established by feeding the animals with high fat diet for 8 weeks. EAS was applied to bilateral ＂Pishu＂ （BL 20）, ＂Geshu＂ （BL 17） and ＂Shenshu＂ （BL 23） for 20 min, once daily for 4 weeks. Rats of the medication group were treated by 1% Dongbao Gantai suspension （0.28 g/kg,20 mL/kg） once daily for 4 weeks. Pathological changes of the liver tissue were observed by microscope after H.E. staining. Hepatic free fatty acid （FFA） content was assayed by using an automatic biochemistry analyzer, malondialdehyde （MDA） content and superoxide dismutase （SOD） ac- tivity were detected by penthiobarbituric acid colorimetric method and xanthine oxidase colorimetric method, respectively. The expression of liver TNF-α was detected by immunohistochemistry. Results Compared with the normal group, rats of the model group showed a moderate to severe fatty degeneration of liver cells, significant up-regulation of hepatic TNF-α expression, FFA and MDA contents （P〈0. 01）, and marked down-regulation of SOD activity （P〈0.01）. Following 4 weeks＇ treatment, compared with the model group, liver fatty degeneration was reduced at different degrees in both EAS and medication groups; liverFFA and MDA contents and TNF-α expression were significantly down-regulated （P〈0.01, P〈0.05）, and hepatic SOD activity was notably increased （P〈0.01, P〈0.05） in both EAS and medication groups, suggesting a reduction of hepatic lipid peroxidation. No significant differences between the EAS and medication groups in the liver FFA and MDA contents, SOD activity and TNF- α expression （P〈0.05）. Cenclusion EA intervention can improve liver fatty degeneration, inhibit high fat induced up-regulation of hepatic TNF-α expression, FFA and MDA contents and down-regulation of SOD activity in non-alcohol fatty liver model rats, which may contribute to its effect in improving NAFLD.