冠心康对动脉粥样硬化LDLR^(-/-)小鼠肝组织MAPKs/NF-κB信号通路相关蛋白表达的影响

Effect of Guanxinkang on expression of proteins related to MAPKs/NF-κB signal pathway in liver tissue of LDLR^(-/-) mice with atherosclerosis

目的观察冠心康对LDLR^(-/-)小鼠肝组织MAPKs/NF-κB信号通路相关蛋白表达的影响。方法将24只雄性LDLR^(-/-)小鼠随机分为模型组,冠心康低、中、高剂量组,以C57BL/6J小鼠为对照组。除对照组外,每组给予高脂饲料喂养12周。对照组、模型组每日给予200μL 0.9%NaCl溶液灌胃,冠心康低、中、高剂量组分别每日给予7.24 g/kg、14.48 g/kg、28.96 g/kg的中药煎剂200μL灌胃,共12周。采用生化检测方法检测小鼠血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、总胆固醇(TC)、三酰甘油(TG)水平;油红O染色法观察小鼠斑块面积;HE染色法观察小鼠肝组织结构;Western blot测定细胞外调节蛋白激酶(Erk1/2),分子量为38 kDa的促分裂素愿活化蛋白激酶(P38),c-Jun氨基末端激酶(JNK),核因子κB抑制蛋白(IκB),核因子κB蛋白表达(NF-κB);RT-PCR、ELISA法检测白介素-1β(IL-1β)、白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)mRNA及含量;免疫荧光法观察TNF-α表达。结果与模型组比较,冠心康各剂量组小鼠血清TC、TG、ALT、AST水平降低(P<0.05),主动脉斑块面积减少,肝组织病理变化改善,肝组织Erk1/2,P38,JNK,IκB,NF-κB水平降低(P<0.05),肝组织IL-1β、IL-6、TNF-αmRNA表达下调(P<0.05),血清IL-1β、IL-6、TNF-α含量减少(P<0.05),肝组织TNF-α表达减少(P<0.05)。结论冠心康可通过抑制MAPKs/NF-κB信号通路,减轻LDLR^(-/-)小鼠肝脏炎症,进而改善动脉粥样硬化。

Objective To observe the effect of Guanxinkang(GXK) on the expression of proteins related to MAPKs/NF-κB signal pathway in the liver tissue of LDLR^(-/-) mice.Methods Twenty-four male LDLR^(-/-) mice were randomly divided into model group,GXK low,medium and high dose groups,and C57 BL/6 J mice were used as the control group. Except for the control group,each group was given high-fat feed for 12 weeks. The control group and the model group were given 200 μL of 0.9% NaCl solution daily. The GXK low,medium and high dose groups were given 7.24 g/kg,14.48 g/kg,28.96 g/kg of Chinese medicine decoction,200 μL daily for 12 weeks. The serum levels of TC,TG,ALT and AST were detected by biochemical assay;the plaque area was observed by oil red O staining;the liver tissue structure was observed by hematoxylin-eosin(HE) staining;the expressions of Erk1/2,P38,JNK,IκB,NF-κB proteins were determined by Western blot;the mRNA and contents of IL-1,IL-6 and TNF-α were detected by RTPCR and ELISA respectively;and the expression of TNF-α was observed by immunofluorescence.Results Compared with the model group,the levels of serum TC,TG,ALT and AST were decreased in each dose group of GXK(P<0.05);the area of aortic plaques was reduced and pathological changes in the liver tissue were improved. And the phosphorylation level of Erk1/2,P38,JNK,IκB,and NF-κB protein in liver tissue was decreased in GXK groups(P<0.05). The expressions of IL-1β,IL-6,TNF-α mRNA in the liver tissue were down-regulated in GXK groups(P<0.05). The contents of IL-1β,IL-6 and TNF-α in serum were decreased(P<0.05)and the expression of TNF-α in liver tissue was reduced.Conclusion GXK can reduce liver inflammation and improve atherosclerosis in LDLR^(-/-) mice by inhibiting MAPKs/NF-κB signal pathway.