黄连解毒汤调节Akt/mTOR及Beclin-1自噬信号通路改善SHR主动脉损伤作用机制的研究

Mechanism of Huanglian Jiedu Decoction(黄连解毒汤)Regulating Akt/mTOR and Beclin-1 Autophagy Signaling Pathway to Improve SHR Aortic Injury

目的基于Akt/mTOR及Beclin-1自噬信号通路探讨黄连解毒汤(Huanglian Jiedu Decoction,HLJDD)对自发性高血压病大鼠(spontaneous hypertension rat,SHR)主动脉损伤改善作用的研究。方法将50只SHR随机分为模型组、HLJDD低剂量组(75 mg·kg^(-1)·d^(-1))、中剂量组(150 mg·kg^(-1)·d^(-1))、高剂量组(300 mg·kg^(-1)·d^(-1))以及卡托普利组(0.175 mg·kg^(-1)·d^(-1))5个组,另设WKY大鼠为空白对照组,每组10只。模型组和空白对照组灌胃给予等体积的氯化钠溶液,每天灌胃给药1次,5 d 1个疗程,共8个疗程。实验末,取主动脉HE染色检测组织病理学变化,TUNEL染色检测主动脉细胞凋亡率,采用qRT-PCR和Western blot检测Akt/mTOR及Beclin-1自噬通路相关基因和蛋白表达水平变化。结果与空白对照组相比,模型组主动脉内膜脱落严重,中膜弹性纤维排列发生紊乱,细胞凋亡率显著升高,HLJDD中、高剂量组和卡托普利组能够有效改善组织病理现象,显著降低细胞凋亡率。与空白对照组相比,模型组中Akt、mTOR、Beclin-1基因表达水平以及mTOR、Akt、LC3-II及Beclin-1的蛋白表达水平显著升高,经HLJDD干预后相关基因和蛋白表达水平均有不同程度的降低。结论HLJDD能够有效改善SHR主动脉组织损伤,其机制可能与抑制Akt/mTOR及Beclin-1自噬相关信号通路有关。

Objective To investigate the improvement of Huanglian Jiedu Decoction(黄连解毒汤,HLJDD)on aortic injury in spontaneously hypertensive rats(SHR)based on Akt/mTOR and beclin-1 autophagy signal pathway.Methods Fifty SHRs were randomly divided into five groups:model group,HLJDD low dose group(75 mg·kg^(-1)·d^(-1)),medium dose group(150 mg·kg^(-1)·d^(-1)),high dose group(300 mg·kg^(-1)·d^(-1))and captopril group(0.175 mg·kg^(-1)·d^(-1)).WKY rats were set as blank control group,with 10 rats in each group.The model group and the blank group were given the same volume of normal saline by gavage,once a day,a course of treatment for 5 days,a total of 8 courses of treatment.At the end of the experiment,HE staining was used to detect the histopathological changes.TUNEL staining was used to detect the apoptosis rate of aortic cells,and QRT PCR and Western blot were used to detect the expression levels of Akt/mTOR and beclin-1 autophagy pathway related genes and proteins.Results Compared with those of the blank control group,the aortic intima in the model group fell off seriously,the arrangement of elastic fibers in the middle membrane was disordered,and the apoptosis rate increased significantly.The medium and high dose HLJDD groups and captopril group can effectively improve the histopathological phenomenon and significantly reduce the apoptosis rate.Compared with those of the blank control group,the gene expression levels of Akt,mTOR and beclin-1 and the protein expression levels of mTOR,Akt,LC3-II and beclin-1 in the model group were significantly increased,and the expression levels of related genes and proteins decreased in varying degrees after HLJDD intervention.Conclusion HLJDD can effectively improve aortic tissue injury in SHR,and its mechanism may be related to the inhibition of Akt/mTOR and beclin-1 autophagy related signal pathway.