番石榴叶水提物对四氧嘧啶诱导体外胰岛INS-1细胞凋亡的保护作用及其机制  

Protective effect and mechanism of water extract from Psidium guajava leaves on alloxan-induced apoptosis of INS-1 cells

作　　者：梁结斐 陈震尧 吴伟斌[3] LIANG Jiefei;CHEN Zhenyao;WU Weibin(Department of Pharmacy,Zhaoqing Medical College,Guangdong Province,Zhaoqing 526020,China;Department of Pharmacy,Zhaoqing Hospital of Traditional Chinese Medicine,Guangdong Province,Zhaoqing 526020,China;Department of Basic Medical Science,Zhaoqing Medical College,Guangdong Province,Zhaoqing 526020,China)

机构地区：[1]肇庆医学高等专科学校药学院,广东肇庆526020 [2]广东省肇庆市中医院药剂科,广东肇庆526020 [3]肇庆医学高等专科学校基础医学院,广东肇庆526020

出　　处：《中国当代医药》2022年第31期5-10,15,共7页China Modern Medicine

基　　金：广东省中医药局中医药科研项目(20211456);广东省肇庆市科技创新指导类项目(2021040311010)。

摘　　要：目的探讨番石榴叶(PGL)水提物对四氧嘧啶(ALX)诱导的体外INS-1细胞凋亡的保护作用及其可能的机制。方法以大鼠胰岛细胞瘤细胞株INS-1建立体外模型,利用CCK-8法测定药物对INS-1细胞活力的影响;采用化学自发光法测定各组细胞的细胞内ATP含量;采用Hoechst33342法测定凋亡形态学;采用Annexin V-FITC法测定细胞凋亡;采用DCFH-DA法测定细胞内活性氧含量;采用Rhodamin 123法检测细胞线粒体膜电位;采用Western blot法测定Bcl-2、Bax、Cleaved-Caspase 3、GRP78、CHOP、Nrf2、HO-1蛋白的表达量。结果PGL水提物各剂量组的细胞活力、细胞内ATP含量、线粒体膜电位及Bcl-2、Nrf2及HO-1等凋亡及抗氧化应激相关蛋白的表达量和Nrf2的核转位程度均高于ALX20组,差异有统计学意义(P<0.05);PGL水提物各剂量组的细胞核浓染、核固缩及凋亡细胞的比例、细胞内活性氧含量以及Bax、Cleaved-Caspase 3、GRP78及CHOP等凋亡及内质网应激相关蛋白的表达均低于ALX组,差异有统计学意义(P<0.05)。结论PGL水提物能有效逆转ALX诱导的体外INS-1细胞凋亡,其机制与PGL水提物增强细胞抗氧化应激、减轻内质网应激有关。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.

关 键 词：番石榴叶 INS-1细胞 氧化应激 内质网应激 

分 类 号：R285.5[医药卫生—中药学]