基于Nrf2/Keap1/HO-1信号通路探究针刺疗法对高脂血症大鼠氧化应激与脂质水平的影响

Effect of acupuncture therapy on oxidative stress and lipid levels in rats with hyperlipidemia based on the Nrf2/Keap1/HO-1 signaling pathway

目的基于Nrf2/Keap1/HO-1信号通路探究针刺疗法对高脂血症大鼠氧化应激与脂质水平的改善作用。方法SPF级雄性Wistar大鼠32只,随机分成空白组、模型组、辛伐他汀组(20 mg·kg^(-1))和针刺组,每组8只。空白组饲喂普通饲料,自由饮用蒸馏水;其他各组饲喂高脂饲料,自由饮用10%(m·v^(-1))果糖溶液方法建立高脂血症大鼠模型,共8 w。采用酶联免疫吸附法(ELISA)测定大鼠血清TC、TG、LDL、HDL、AST、ALT、SOD、CAT、MDA水平;HE染色观察肝脏组织病理学改变;蛋白质免疫印迹法(Western Blot,WB)测定Nrf2、HO-1、Keap1蛋白表达。结果与空白组比较,模型组TC、TG、LDL、ALT、AST、MDA水平均明显升高,HDL、SOD、CAT水平明显降低,差异有统计学意义(P<0.05或0.01)。肝脏组织H&E染色切片肝细胞排列略紊乱,出现轻度脂肪病变,Nrf2、HO-1蛋白表达降低,Keap1蛋白表达升高;与模型组相比,针刺组TC、TG、LDL、ALT、AST、MDA均明显降低,HDL、SOD、CAT水平明显升高,差异有统计学意义(P<0.05或0.01)。肝脏组织H&E染色切片脂肪性病变程度较低,Nrf2、HO-1蛋白表达升高,Keap1蛋白表达降低。结论针刺疗法可以通过激活Nrf2/Keap1/HO-1信号通路增加机体抗氧化能力,从而达到治疗高脂血症的目的。

Objective To expolore improvement effects of acupuncture therapy on oxidative stress and lipid levels in rats with hyperlipidemia based on the Nrf2/Keap1/HO-1 signaling pathway.Methods 32 specific pathogen free(SPF)male Wistar rats were randomly divided into the blank group,the model group,the simvastatin group(20 mg·kg^(-1))and the acupuncture group,with 8 rats in each group.The blank group was fed with regular feed and freely drank distilled water;The other groups were fed with high-fat feed and freely drank 10%(m·v^(-1))fructose solution to establish a rat model of hyperlipidemia for a total of 8 weeks.Serum total cholesterol(TC),triglycerie(TG),low-density lipoprotein(LDL),high density lipoprotein(HDL),aspartate transaminase(AST),alanine transaminase(ALT),superoxide dismutase(SOD),CAT and MDA levels in rats were determined by enzyme-linked immunosorbent assay(ELISA);HE staining was used to observe histopathological changes in the liver;Western Blot(WB)was used to determine the expression of Nrf2,HO-1and Keap1 proteins.Results Compared with the blank group,in the model group the levels of TC,TG,LDL,ALT,AST and MDA were significantly increased(P<0.01),and the levels of HDL,SOD and CAT were significantly reduced(P<0.01).The arrangement of liver cells in H&E stained sections of liver tissue was slightly disordered,and mild fatty lesions appeared.The expression of Nrf2 and HO-1 proteins was reduced,and the expression of Keap1 protein was increased.Compared with the model group,in the acupuncture group the levels of TC,TG,LDL,ALT,AST and MDA were significantly reduced(P<0.01),and HDL,SOD and CAT levels were significantly increased(P<0.01).The degree of fatty lesions in H&E stained sections of liver tissue was lower.The expression of Nrf2 and HO-1 proteins was increased,and the expression of Keap1 protein was reduced.Conclusion Acupuncture therapy can increase body antioxidant capacity by activating the Nrf2/Keap1/HO-1 signaling pathway,thereby achieving the purpose of treating hyperlipidemia.