Effects of autophagy inhibitor 3-methyladenine on a diabetic mice model

作者全名："Ren, Hai-Wen; Yu, Wen; Wang, Ya-Nan; Zhang, Xin-Yi; Song, Shun-Qiong; Gong, Shu-Yu; Meng, Ling-Yao; Gan, Chen; Liu, Ben-Ju; Gong, Quan"

作者地址："[Ren, Hai-Wen; Yu, Wen; Wang, Ya-Nan; Zhang, Xin-Yi; Song, Shun-Qiong; Gong, Shu-Yu; Meng, Ling-Yao; Gan, Chen] Yangtze Univ, Hlth Sci Ctr, Jingzhou 434023, Hubei, Peoples R China; [Ren, Hai-Wen] Chongqing Med Univ, Bishan Hosp, Dept Clin Lab, Chongqing 402760, Peoples R China; [Liu, Ben-Ju] Yangtze Univ, Med Sch, Dept Human Anat, Jingzhou 434023, Hubei, Peoples R China; [Gong, Quan] Yangtze Univ, Clin Mol Immunol Ctr, Med Sch, Jingzhou 434023, Hubei, Peoples R China; [Gong, Quan] Yangtze Univ, Med Sch, Dept Immunol, Jingzhou 434023, Hubei, Peoples R China"

通信作者："Liu, BJ (通讯作者)，Yangtze Univ, Med Sch, Dept Human Anat, Jingzhou 434023, Hubei, Peoples R China.; Gong, Q (通讯作者)，Yangtze Univ, Med Sch, Dept Immunol, Jingzhou 434023, Hubei, Peoples R China."

来源：INTERNATIONAL JOURNAL OF OPHTHALMOLOGY

ESI学科分类：CLINICAL MEDICINE

WOS号：WOS:001067764300009

JCR分区：Q2

影响因子：1.9

年份：2023

卷号：16

期号：9

开始页：1456

结束页：1464

文献类型：Article

关键词：diabetic mellitus; 3-methyladenine; autophagy; fibrosis; apoptosis; mice

摘要："AIM: To investigate the role of autophagy inhibitor 3-methyladenine (3-MA) on a diabetic mice model (DM) and the potential mechanism. METHODS: Male C57BL/6J mice were randomly divided into a normal control group (NC group) and an DM group. DM were induced by multiple low-dose intraperitoneal injection of streptozotocin (STZ) 60 mg/kg.d for 5 consecutive days. DM mice were randomly subdivided into untreated group (DM group), 3-MA (10 mg/kg. d by gavage) treated group ( DM+3-MA group) and chloroquine (CQ; 50 mg/kg by intraperitoneal injection) treated group (DM+CQ group). The fasting blood glucose (FBG) levels were recorded every week. At the end of experiment, retinal samples were collected. The expression levels of pro-apoptotic proteins cleaved caspase-3, cleaved poly ADP-ribose polymerase 1 (PARP1) and Bax, anti-apoptotic protein Bcl- 2, fibrosis-associated proteins Fibronectin and type 1 collagen a1 chain (COL1A1), vascular endothelial growth factor (VEGF), inflammatory factors interleukin (IL)-1 beta and tumor necrosis factor (TNF)-a, as well as autophagy related proteins LC3, Beclin-1 and P62 were determined by Western blotting. The oxidative stress indicators 8- hydroxydeoxyguanosine (8-OHdG) and malondialdehyde (MDA) were detected by commercial kits. RESULTS: Both 3- MA and CQ had shor t- term hypoglycemic effect on FBG and reduced the expression of VEGF and inflammatory factors IL-1 beta and TNF-alpha in DM mice. 3-MA also significantly alleviated oxidative stress indicators 8-OHdG and MDA, decreased the expression of fibrosisrelated proteins Fibronectin and COL1A1, pro-apoptotic proteins cleaved caspase- 3, cleaved PARP1, as well as the ratio of Bax/Bcl-2. CQ had no significant impact on the oxidative stress indicators, fibrosis, and apoptosis related proteins. The results of Western blotting for autophagy related proteins showed that the ratio of LC3 II/LC3 I and the expression of Beclin-1 in the retina of DM mice were decreased by 3-MA treatment, and the expression of P62 was further increased by CQ treatment. CONCLUSION: 3-MA has anti-apoptotic and anti-fibrotic effects on the retina of DM mice, and can attenuate retinal oxidative stress, VEGF expression and the production of inflammatory factors in the retina of DM mice. The underlying mechanism of the above effects of 3-MA may be related to its inhibition of early autophagy and hypoglycemic effect."

基金机构：National Natural Science Foundation of China [82270893]; Joint Fund of Health Committee of Hubei Province [WJ2019-16]

基金资助正文：Supported by the National Natural Science Foundation of China (No.82270893); Joint Fund of Health Committee of Hubei Province (No.WJ2019-16).