田黄方对高脂饮食诱导小鼠肾脏损伤的改善作用及机制研究

Effect and mechanism of Tianhuang Formula on high fat diet induced renal injury in mice

目的探究田黄方(THF)对高脂饮食诱导C57BL/6J小鼠肾脏损伤的改善作用及机制。方法将SPF级的C57BL/6J雄性小鼠分为正常组、模型组、田黄方组,高脂饮食诱导5个月及给药干预6周;采用H&E染色观察小鼠肾脏组织病理情况;生化检测试剂盒检测肾脏匀浆脂代谢和氧化应激状态指标:乳酸脱氢酶(LDH)、低密度脂蛋白胆固醇(LDL-C)、总胆固醇(TC)、三酰甘油(TG)、丙二醛(MDA)和超氧化物歧化酶(SOD)水平;末端标记法(TUNEL)染色检测小鼠肾脏组织细胞凋亡情况;免疫组化法检测小鼠肾脏组织中凋亡蛋白Cleaved caspase-3的原位表达情况;采用蛋白免疫印迹法(WB)法检测PI3K、p-AKT、Bax、Cleaved caspase-3和Bcl-2蛋白的表达水平。结果与正常组相比,模型组小鼠肾脏病理可见肾小球体积增大及球囊黏连,肾脏TC、TG、LDL-C、LDH和MDA水平显著升高,SOD水平显著降低,凋亡蛋白Bax、Cleaved caspase-3蛋白表达亦显著增加,抗凋亡通路蛋白PI3K、pAKT、Bcl2和Bcl2/Bax表达显著减少。与模型组相比,THF组可有效改善肾脏病理损伤,并显著抑制高脂诱导的凋亡信号通路的激活。结论THF能够有效改善高脂诱导的小鼠肾损伤,其机制可能与调节PI3K/AKT信号通路、抑制细胞凋亡、改善机体氧化应激及脂质代谢紊乱有关。

Objective To explore the effect and mechanism of Tianhuang Formula(THF) on renal injury induced by high fat diet in C57BL/6J mice. Methods SPF-grade C57BL/6J male mice were divided into control group,model group and THF group, and high fat diet was fed for 5 months and drug intervention was carried out for 6weeks. H&E staining was used to observe the pathological conditions of kidney of mice. The levels of LDH, LDL-C,TC, TG, MDA and SOD in kidney were detected by biochemical assay kit to evaluate the lipid metabolism and oxidative stress levels. TUNEL staining was used to observe apoptosis cells in kidney. The expression of Cleaved caspase-3 in kidney was detected by immunohistochemical assay. The protein expression levels of PI3K, p-AKT,Bax, Cleaved caspase-3 and Bcl-2 were detected by Western blotting. Results Compared with the control group,The kidney showed increased giomerular volume, balloon adhesions, decreased SOD, and the increased levels of TC, TG, LDL-C, LDH and MDA significantly in model group. In the model group, the expression levels of Bax and Cleaved caspase-3 were up-regulated significantly, and PI3K, p-AKT, Bcl2, Bcl2/Bax were down-regulated markedly. Compared with the model group, THF could effectively alleviate the renal pathological damage and inhibit the activation of apoptosis signaling pathway induced by high fat diet in THF group. Conclusion THF could effectively improve the renal injury induced by high fat diet in mice, and the mechanism may be related to the regulation of PI3K/AKT signaling pathway, inhibition of cell apoptosis, alleviation of oxidative stress and lipid metabolism disorder.