Liang Bing, Wei Wei, Wang Jianta, Zhang Mingming, Xu Ran, Wu Fei, Xiao Haitao, Tang Lei
a Department of Pharmacology, School of Basic Medical Science , Guiyang Medical University , Guiyang , China ;
b School of Pharmacy , Guiyang Medical University , Guiyang , China ;
Pharm Biol. 2016 Sep;54(9):1656-63. doi: 10.3109/13880209.2015.1113993. Epub 2016 Mar 14.
Context Hydrogen peroxide (H2O2)-induced damage in the lens epithelium leads to cell death and cataract. Semiaquilegia adoxoides (DC.) Makino (Ranunculaceae), a folk medicine of Hmong (an ethnic group of China), has been traditionally used to treat cataract; however, the underlying molecular mechanism is yet to be uncovered. Objective This study aimed to investigate whether the n-butanol extract of S. adoxoides (nSA) is effective against the H2O2-induced oxidative stress in human lens epithelial (HLE) cells. Materials and methods Human lens epithelial (SRA 01/04) cells were stimulated by H2O2 (250 μM) in the presence or absence of nSA. The antioxidant effects of nSA were determined in terms of cell viability (MTT assay), apoptosis (AnnexinV/PI staining), radical scavenging capability (various enzymatic assays), loss of mitochondrial membrane potential (Rhodamine 123 staining), expression of apoptotic markers including caspase-3 and caspase-9 and the change of Bcl-2/Bax ratio (western blot) in the HLE cells. Results The results showed that pretreatment of nSA (250, 500 and 1000 μg/mL) markedly reduced H2O2-induced cellular apoptosis and malondialdehyde accumulation, but elevated the activities of total superoxide dismutase, catalase, glutathione peroxidase. Thus, the total antioxidative capability was enhanced upon the nSA treatment meanwhile the loss of mitochondrial membrane potential was prevented. Moreover, nSA at concentrations of 250, 500 and 1000 μg/mL also significantly suppressed the activation of caspase-3 and -9, and increased the Bcl-2/Bax ratio in the HLE cells. Discussion and conclusion Our findings suggested that nSA is a potential prophylactic agent in the prevention of cataractogeneis.
背景 过氧化氢(H2O2)诱导的晶状体上皮损伤会导致细胞死亡和白内障。伏毛铁棒锤(毛茛科)是中国苗族的一种民间药物,传统上用于治疗白内障;然而,其潜在的分子机制尚未被揭示。目的 本研究旨在探讨伏毛铁棒锤正丁醇提取物(nSA)是否对H2O2诱导的人晶状体上皮(HLE)细胞氧化应激有效。材料与方法 在有或无nSA存在的情况下,用H2O2(250μM)刺激人晶状体上皮(SRA 01/04)细胞。通过细胞活力(MTT法)、凋亡(AnnexinV/PI染色)、自由基清除能力(各种酶法检测)、线粒体膜电位丧失(罗丹明123染色)、凋亡标志物包括半胱天冬酶-3和半胱天冬酶-9的表达以及HLE细胞中Bcl-2/Bax比值的变化(蛋白质免疫印迹法)来确定nSA的抗氧化作用。结果 结果表明,nSA(250、500和1000μg/mL)预处理显著降低了H2O2诱导的细胞凋亡和丙二醛积累,但提高了总超氧化物歧化酶、过氧化氢酶、谷胱甘肽过氧化物酶的活性。因此,nSA处理后总抗氧化能力增强,同时线粒体膜电位丧失得到预防。此外,250、500和1000μg/mL浓度的nSA也显著抑制了HLE细胞中半胱天冬酶-3和-9的激活,并增加了Bcl-2/Bax比值。讨论与结论 我们的研究结果表明,nSA是预防白内障发生的一种潜在预防剂。