基于AMPK-PGC-1α信号通路探讨“上火”动物模型的发病机制

Study on the pathogenesis of the animal model of‘Shanghuo’based on AMPK-PGC-1αsignal pathway

目的:探讨AMPK-PGC-1α信号通路及氧化应激在“上火”动物模型中的作用。方法:用干姜附子肉桂水煎液灌胃构建阴虚动物模型,用党参黄芪水煎液灌胃构建实热动物模型,21d后氢氧化钠晶体灼烧法叠加口腔溃疡模型。记录大鼠体质量、肛温、溃疡愈合等情况;检测血浆中SOD活力和GSH、MDA含量;分别检测肝脏中AMPK、PGC-1αmRNA和蛋白表达水平;HE染色观察口腔黏膜病理学改变。结果:与正常对照组比较,模型组GSH浓度显著升高(P<0.01),阴虚溃疡组浓度显著高于模型组和实热溃疡组(P<0.01);与正常对照组比较,模型组SOD活力显著降低(P<0.01),阴虚溃疡组显著低于模型组和实热溃疡组(P<0.01,P<0.05)。与模型组比较,阴虚溃疡组与实热溃疡组AMPK及PGC-1α基因表达显著下调(P<0.01),且实热溃疡组表达较阴虚溃疡组显著下调(P<0.01),阴虚溃疡组与实热溃疡组AMPK及PGC-1α蛋白表达呈下调趋势,但差异无统计学意义。与阴虚溃疡组比较,实热溃疡组AMPK蛋白表达显著升高(P<0.01)。与正常对照组比较,各组大鼠口腔黏膜组织均发生表皮脱落、萎缩和炎性细胞浸润。结论:“上火”动物模型中存在氧化应激损伤,且实热溃疡模型损伤程度重于阴虚溃疡模型;“上火”动物模型中AMPK-PGC-1α信号通路受到抑制,可能是不同“上火”状态出现抗氧化能力差异的原因之一。

Objective:Researching the role of AMPK-PGC-1αsignaling pathway and oxidative stress in animal models of Shanghuo.Methods:Ganjiang Fuzi Rougui water decoction was used to build yin deficiency animal model.Dangshen Huangqi water decoction was used to build excess heat animal model.The body mass,anal temperature and ulcer healing of rats were recorded;the SOD activity and GSH,MDA content in plasma were detected;the mRNA and protein expression levels of AMPK and PGC-1αin liver were detected respectively;the pathological changes of oral mucosa were observed by HE staining.Results:Compared with normal control group,the GSH concentration in the model group was higher and YD-RAU group was higher than that in the model group and EH-RAU group.Results:Compared with NC group,the SOD of M group was lower(P<0.01)and YD-RAU group was much lower than that in M group and EH-RAU group(P<0.01,P<0.05).Compared with M group,the mRNA level of AMPK and PGC-1αwas much lower in YD-RAU group and EH-RAU group(P<0.01),and the level in EH-RAU group was much lower than the level in YD-RAU group(P<0.01).Compared with M group,the protein level of AMPK and PGC-1αin YD-RAU group and EH-RAU group revealed the down trend but have no statistical differences.Compared with YD-group,the protein level of AMPK was increased in EH-group(P<0.01).The pathological results of oral mucosa of rats showed that all mucosal tissues were infiltrated by inflammatory cells and the epidermal was exfoliation and atrophy.Conclusion:There is oxidative stress injury in the‘Shanghuo’animal model,and the degree of injury in the real heat ulcer model is more serious than that in the yin deficiency ulcer model;the inhibition of ampk-pgc-1αsignal pathway in the‘Shanghuo’animal model may be one of the reasons for the difference of antioxidant capacity in different‘Shanghuo’states.