Protective effect and mechanism of water extract from Psidium guajava leaves on alloxan-induced apoptosis of INS-1 cells
LIANG Jiefei1 CHEN Zhenyao2 WU Weibin
1.Department of Pharmacy,Zhaoqing Medical College,Guangdong Province,Zhaoqing 526020,China;
2.Department of Pharmacy,Zhaoqing Hospital of Traditional Chinese Medicine,Guangdong Province,Zhaoqing 526020,China;
3.Department of Basic Medical Science,Zhaoqing Medical College,Guangdong Province,Zhaoqing 526020,China
Abstract：Objective To investigate the protective effect of water extracts from Psidium guajava leaves (PGL) on INS-1 cells apoptosis induced by alloxan (ALX)in vitro and its possible mechanism.Methods Rat islet cell line INS-1 was used to establish an in vitro model,and the effects of drugs on the viability of INS-1 cells were determined by CCK-8 method.The intracellular ATP content of each group was determined by chemical self-illumination method.Hoechst33342 method was used to determine the apoptotic morphology.Annexin V-FITC assay was used to determine cell apoptosis.The content of intracellular reactive oxygen species was determined by DCFH-DA method.Rhodamin 123 method was used to detect the mitochondrial membrane potential.The protein expression levels of Bcl-2,Bax,Cleaved Caspase 3,GRP78,CHOP,Nrf2 and HO-1 were determined by Western blot.Results The cell viability,intracellular ATP content,mitochondrial membrane potential,expression levels of Bcl-2,Nrf2,HO-1 and the degree of nuclear transposition of Nrf2 in PGL water extract groups were higher than those in ALX20 group,the differences were statistically significant (P＜0.05).The ratio of nuclear pyknosis and apoptotic cells,intracellular reactive oxygen species content and the expression of Bax,Cleaved-Caspase 3,GRP78 and CHOP in PGL water extract groups were lower than those of ALX20 group,the differences were statistically significant (P＜0.05).Conclusion PGL water extracts can effectively reversed the apoptosis of INS-1 cells induced by alloxan in vitro,and the mechanism was related to the enhancement of antioxidant stress and reduction of endoplasmic reticulum stress.
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