基于AMPK/PGC1α信号通路探究降脂逐瘀汤对动脉粥样硬化大鼠脂代谢的影响

Impact of Jiangzhi Zhuyu Decoction on lipid metabolism in atherosclerotic rats based on AMPK/PGC1αsignaling pathway

目的:基于AMPK/PGC1α信号通路探究降脂逐瘀汤对动脉粥样硬化(AS)大鼠脂代谢的影响。方法:高脂饲料喂养及腹腔注射维生素D3诱导建立AS大鼠模型,随机分为模型组、降脂逐瘀汤(3.75 g/kg)低剂量组、降脂逐瘀汤(7.5 g/kg)中剂量组、降脂逐瘀汤(15 g/kg)高剂量组、降脂逐瘀汤(15 g/kg)高剂量+盐酸多索啡(AMPK抑制剂,0.2 mg/kg)组,每组12只,另取12只正常SD大鼠喂养普通饮食及腹腔注射生理盐水作为对照组。经降脂逐瘀汤和盐酸多索啡分组干预处理各组大鼠后,检测大鼠甘油三酯(TG)、总胆固醇(TC)、高密度脂蛋白胆固醇(HDL-C)和低密度脂蛋白胆固醇(LDL-C)水平;HE染色检测大鼠主动脉组织病理变化;油红O染色检测各组大鼠主动脉和肝脏脂质沉积;酶标仪测定大鼠血清炎症因子IL-8、IL-1β、IL-18水平;免疫印迹法检测大鼠主动脉组织AMPK/PGC1α通路蛋白表达。结果:与对照组相比,模型组大鼠主动脉组织发生严重病理损伤,主动脉和肝脏组织中有大量脂质沉积,肝脏脂肪变性严重,血清LDL-C、TC、TG、IL-8、IL-1β、IL-18水平显著升高(P<0.05),血清HDL-C水平、主动脉组织p-AMPK/AMPK与PGC1α蛋白表达显著降低(P<0.05)。与模型组相比,降脂逐瘀汤各剂量组大鼠主动脉组织病理损伤、主动脉脂质沉积和肝脏脂肪变性症状均减轻,血清LDL-C、TC、TG、IL-8、IL-1β、IL-18水平均降低(P<0.05),血清HDL-C水平、主动脉组织p-AMPK/AMPK与PGC1α蛋白表达均升高(P<0.05),且呈剂量依赖性(P<0.05);与降脂逐瘀汤高剂量组相比,降脂逐瘀汤高剂量+盐酸多索啡组大鼠主动脉组织病理损伤、主动脉脂质沉积和肝脏脂肪变性症状加重,血清LDL-C、TC、TG、IL-8、IL-1β、IL-18水平升高(P<0.05),血清HDL-C水平、主动脉组织p-AMPK/AMPK与PGC1α蛋白表达降低(P<0.05)。结论:降脂逐瘀汤可通过激活AMPK/PGC1α信号抵抗炎症发生发展,减轻大鼠主动脉组织病理损伤、主动脉脂质沉积和肝脏脂肪变性症状,改善AS大鼠脂代谢。

Objective:To explore impact of Jiangzhi Zhuyu Decoction on lipid metabolism in atherosclerotic(AS)rats based on AMPK/PGC1αsignaling pathway.Methods:AS model rats were induced and established by high-fat diet feeding and intraperitoneal injection of vitamin D3,and randomly grouped into model group,low-dose Jiangzhi Zhuyu Decoction(3.75 g/kg)group,medium-dose Jiangzhi Zhuyu Decoction(7.5 g/kg)group,high-dose Jiangzhi Zhuyu Decoction(15 g/kg)group,high-dose Jiangzhi Zhuyu Decoction(15 g/kg)+doxorphine hydrochloride(AMPK inhibitor,0.2 mg/kg)group,with 12 rats in each group,another 12 normal SD rats were fed with normal diet and intraperitoneally injected with normal saline as control group.After Jiangzhi Zhuyu Decoction and doxorphine hydrochloride were grouped to intervene rats in each group,levels of triacylglycerol(TG),total cholesterol(TC),high-density lipoprotein cholesterol(HDL-C)and low-density lipoprotein cholesterol(LDL-C)in rats were measured;HE staining was used to detect pathological changes of rat aorta;oil red O staining was used to detect lipid deposition in aorta and liver of rats in each group;microplate reader was used to detect levels of serum inflammatory mediators IL-8,IL-1βand IL-18;Western blot was used to detect expression of AMPK/PGC1αpathway protein in rat aorta.Results:Compared with control group,model group had severe pathological damage to aorta tissue,a large amount of lipid deposition in aorta and liver tissue,and severe liver steatosis,serum LDL-C,TC,TG,IL-8,IL-1βand IL-18 levels were obviously increased(P<0.05),serum HDL-C level,aortic tissue p-AMPK/AMPK and PGC1αprotein expression were obviously decreased(P<0.05).Compared with model group,aortic tissue pathological damage,aortic lipid deposition and hepatic steatosis symptoms of rats in Jiangzhi Zhuyu Decoction groups were alleviated,serum LDL-C,TC,TG,IL-8,IL-1βand IL-18 were obviously decreased(P<0.05),serum HDL-C level,aortic tissue p-AMPK/AMPK and PGC1αprotein expressions were obviously increased(P<0.05),which in a dose-dependent manner(P<0.05);compared with high-dose Jiangzhi Zhuyu Decoction group,aortic tissue pathological damage,aortic lipid deposition and liver steatosis were aggravated in high-dose Jiangzhi Zhuyu Decoction+doxorphine hydrochloride group,serum LDL-C,TC,TG,IL-8,IL-1β,and IL-18 levels were obviously increased(P<0.05),serum HDL-C level,aortic tissue p-AMPK/AMPK and PGC1αprotein expressions were obviously decreased(P<0.05).Conclusion:Jiangzhi Zhuyu Decoction can resist occurrence and development of inflammation by activating AMPK/PGC1αsignal,alleviate aortic pathological damage,aortic lipid deposition and liver steatosis symptoms in rats,and improve lipid metabolism in AS rats.