Huo Hai-Yan, Wang Jin, Zhang Xu-Mei, Zhang Wen-Ting, Yue Ji-Ping, Jiao Xiang-Ying
Departement of Physiology, Shanxi Medical University, Taiyuan 030001, China.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2020 Mar;36(2):119-123. doi: 10.12047/j.cjap.5878.2020.027.
To investigate whether the increased expression of thioredoxin interacting protein (TXNIP) in diabetes affects the senescence of islet β cells. Six normal mice (db/m) and six diabetic mice (db/db) were randomly selected. Fasting blood glucose was measured by blood sugar meter, the expression levels of TXNIP protein, p16, p21 and Rb in pancreatic tissues were detected by Western blot, senescence-associated beta-galactosidase activity in pancreatic tissue was determined by immunochemical staining. INS-1 islet beta cells were randomly divided into 7 groups (=6), and transfected with lentiviruses (30 μl) for 4 to 6 hours, then was screened with puromycin (PM, 3 μg/m) for 7 days to construct normal group, scramble ShRNA group (interference with airborne poison group), TXNIP-ShRNA-1 group (TXNIP silence group-1), TXNIP-ShRNA-2 group (TXNIP silence group 2), TXNIP-ShRNA-3 group (TXNIP silence group 3), Ad-GFP group (overexpression of the air virus group), Ad-TXNIP-GFP group (TXNIP overexpression group) stably transferred INS-1 islet beta cell line. TXNIP protein expression was detected by Western blot, aging-related beta-galactosidase activity was detected by immunochemical staining, the changes of expression of p16, p21 and Rb was determined by Western blot. Compared with normal mice, the fasting blood glucose of db/db group was increased significantly (＜0. 01), the expression of TXNIP protein was increased significantly in pancreatic tissues(＜0. 05), positive staining rate of β- galactosidase was increased significantly in pancreatic tissues, p16/p21/Rb protein expression levels were increased significantly (＜0. 05). Compared with Ad-GFP group, the positive staining rate of β- galactosidase in Ad-TXNIP-GFP group was increased significantly, p16/p21/Rb protein expression levels were increased significantly (＜0. 01). Compared to the scramble ShRNA group, the positive staining rate of β- galactosidase in TXNIP-ShRNA group was decreased, p16/p21/Rb protein expression levels were decreased significantly (＜0. 05). Diabetes can induce islet β-cell senescence by up-regulating TXNIP expression.
为研究糖尿病中硫氧还蛋白相互作用蛋白（TXNIP）表达增加是否影响胰岛β细胞衰老。随机选取6只正常小鼠（db/m）和6只糖尿病小鼠（db/db）。用血糖仪测量空腹血糖，采用蛋白质免疫印迹法检测胰腺组织中TXNIP蛋白、p16、p21和Rb的表达水平，通过免疫化学染色法测定胰腺组织中衰老相关β-半乳糖苷酶活性。将INS-1胰岛β细胞随机分为7组（每组n = 6），用慢病毒（30 μl）转染4至6小时，然后用嘌呤霉素（PM，3 μg/ml）筛选7天，构建正常组、乱序短发夹RNA组（干扰空载病毒组）、TXNIP短发夹RNA-1组（TXNIP沉默组-1）、TXNIP短发夹RNA-2组（TXNIP沉默组2）、TXNIP短发夹RNA-3组（TXNIP沉默组3）、腺病毒绿色荧光蛋白组（空载病毒过表达组）、腺病毒TXNIP绿色荧光蛋白组（TXNIP过表达组）稳定转染的INS-1胰岛β细胞系。采用蛋白质免疫印迹法检测TXNIP蛋白表达，通过免疫化学染色法检测衰老相关β-半乳糖苷酶活性，采用蛋白质免疫印迹法测定p16、p21和Rb表达变化。与正常小鼠相比，db/db组空腹血糖显著升高（P＜0.01），胰腺组织中TXNIP蛋白表达显著增加（P＜0.05），胰腺组织中β-半乳糖苷酶阳性染色率显著升高，p16/p21/Rb蛋白表达水平显著增加（P＜0.05）。与腺病毒绿色荧光蛋白组相比，腺病毒TXNIP绿色荧光蛋白组β-半乳糖苷酶阳性染色率显著升高，p16/p21/Rb蛋白表达水平显著增加（P＜0.01）。与乱序短发夹RNA组相比，TXNIP短发夹RNA组β-半乳糖苷酶阳性染色率降低，p16/p21/Rb蛋白表达水平显著降低（P＜0.05）。糖尿病可通过上调TXNIP表达诱导胰岛β细胞衰老。
