Acetylcholine protects high glucose-induced H9c2 cell injury by inhibiting autophagy / 中国药理学通报

ABSTRACT

Aim To study the effects of acetylcholine (ACh) on high glucose (HG)-induced cardiomyocytes injury. Methods H9c2 cells were treated with HG for 72 h to establish a model of HG-induced cardiomyocyte injury. The experiment was divided into control, HG group, HG + ACh(10~4 nmol • L"1) group, HG + ACh+ AICAR (0.5 nmol • L"1) group, HG + ACh + Rap(50 nmol • L'1) group, and HG+3-MA (2 mmol • L"1 ) group. Cell viability was evaluated by MTT and ATP assay. The expressions of autophagy, apopto-sis and AMPK-mTOR signaling pathway were detected by Western blot. The number of autophagosomes in each group was observed by transmission electron microscopy. Results Compared with control group, the ratio of LC3-II/LC3-I , p-AMPK/AMPK decreased (P < 0. 05), and the ratio of p-mTOR/mTOR and Bax/Bcl-2 increased in HG group (P <0. 05). Compared with HG group, the activity of cardiomyocytes in HG + ACh group increased, the ratio of LC3-D/LC3-I , p-AMPK/AMPK, Bax/Bcl-2 decreased (P < 0.05), the ratio of p-mTOR/mTOR increased (P < 0. 05 ) , and the ratio of Bax/Bcl-2 decreased in HG + 3-MA group. Compared with HG + ACh group, the p-AMPK/AMPK and p-mTOR/mTOR increased in HG + ACh + AICAR group (P < 0. 05 ) , LC3-U/LC3-1 was elevated ( P < 0. 05 ) , and the ratio of Bax/Bcl-2 increased in HG + ACh + Rap group. Conclusions ACh inhibits autophagy through the AMPK signaling pathway to protect HG-induced cardiomyocytes injury.