Fu Jianming, Hu Xiaojie
Department of Ophthalmology, The Second People's Hospital of Yueqing, Wenzhou, Zhejiang 325608, P.R. China.
Department of Ophthalmology, Wenzhou People's Hospital, Wenzhou, Zhejiang 325000, P.R. China.
Exp Ther Med. 2022 Jun;23(6):420. doi: 10.3892/etm.2022.11347. Epub 2022 Apr 29.
Diabetic cataracts (DC) is one of the main causes of blindness among patients with diabetes mellitus. The aim of the present study was to examine the effect of simvastatin on lens epithelial cells in DC and the underlying mechanism. The viability of SRA01/04 cells treated with different concentrations of simvastatin was detected using a Cell Counting Kit-8 assay before and after high glucose (HG) treatment. The expression levels of E-cadherin, N-cadherin, Vimentin and α-smooth muscle actin (α-SMA), proteins associated with epithelial-mesenchymal transition (EMT), in addition to RhoA, Rho-associated kinases (ROCK)1 and ROCK2, proteins related to RhoA/ROCK signaling, were also measured in SRA01/04 cells treated with HG and simvastatin, with or without U46619, using western blot analysis. DCFH-DA dyes, superoxide dismutase (SOD) and glutathione (GSH)/glutathione disulfide (GSSG) kits were used to measure the levels of oxidative stress parameters in SRA01/04 cells treated with HG and simvastatin with or without U46619. The cell viability of SRA01/04 cells treated with simvastatin was found to be significantly elevated after HG treatment. The protein expression levels of E-cadherin were increased but those of N-cadherin, Vimentin and α-SMA decreased after HG and simvastatin treatment, and this was reversed by U46619. The levels of SOD and GSH-GSSG were found to be increased whereas reactive oxygen species levels were decreased, effects that were reversed by U46619. Additionally, the protein expression levels of RhoA, ROCK1 and ROCK2 were markedly decreased. These findings provided evidence that simvastatin increased HG-induced SRA01/04 cell viability and exerted inhibitory effects on EMT and oxidative stress that occurs during DC.
糖尿病性白内障（DC）是糖尿病患者失明的主要原因之一。本研究的目的是探讨辛伐他汀对DC晶状体上皮细胞的影响及其潜在机制。在高糖（HG）处理前后，使用细胞计数试剂盒-8检测不同浓度辛伐他汀处理的SRA01/04细胞的活力。还通过蛋白质印迹分析测量了HG和辛伐他汀处理的SRA01/04细胞中与上皮-间质转化（EMT）相关的蛋白质E-钙黏蛋白、N-钙黏蛋白、波形蛋白和α-平滑肌肌动蛋白（α-SMA）的表达水平，以及与RhoA/ROCK信号相关的蛋白质RhoA、Rho相关激酶（ROCK）1和ROCK2的表达水平，处理条件包括添加或不添加U46619。使用DCFH-DA染料、超氧化物歧化酶（SOD）和谷胱甘肽（GSH）/谷胱甘肽二硫化物（GSSG）试剂盒测量HG和辛伐他汀处理的SRA01/04细胞中氧化应激参数的水平，处理条件包括添加或不添加U46619。发现HG处理后，用辛伐他汀处理的SRA01/04细胞的活力显著提高。HG和辛伐他汀处理后，E-钙黏蛋白的蛋白质表达水平升高，而N-钙黏蛋白、波形蛋白和α-SMA的表达水平降低，且U46619可逆转这种变化。发现SOD和GSH-GSSG的水平升高，而活性氧水平降低，U46619可逆转这些作用。此外，RhoA、ROCK1和ROCK2的蛋白质表达水平明显降低。这些发现证明辛伐他汀可提高HG诱导的SRA01/04细胞活力，并对DC期间发生的EMT和氧化应激产生抑制作用。
