Department of Reproductive Biomedicine, National Institute of Health and Family Welfare, Baba Gang Nath Marg, Munirka, New Delhi 110067, India.
Apoptosis. 2012 Nov;17(11):1131-43. doi: 10.1007/s10495-012-0749-7.
Using siRNA as a tool, the channelization of pathway in H(2)O(2) induced apoptosis of primary Leydig cells was investigated in vitro. Exposure (4 h) to H(2)O(2) (250 μM) induced maximum apoptosis but affected Leydig cell viability significantly. Therefore, expression of apoptotic marker genes, caspase-8, -9, -3 and polyadenosine ribose polymerase was subsequently investigated using the same concentration post 1 h exposure. Incubation with siRNA (20 nM) either for caspase-8 or -9, inhibited their individual expressions by 55-60 % and activity, 50-55 %. The inhibition efficiency using siRNA was comparable with post- or pre-H(2)O(2) treatment of cells. Like siRNA, Eugenia jambolana (100 μg/ml) plant extract too, effectively countered over-expression of all apoptotic marker proteins. Silencing expressions of caspase 8 but not 9 through siRNA leads to a profound inhibition of caspase 3 implying that H(2)O(2) induced Leydig cell apoptosis is preferably channeled through extrinsic and later extending to other pathways.
采用 siRNA 作为工具，研究了体外 H2O2 诱导原代 Leydig 细胞凋亡过程中信号通路的改变。暴露于 250μM H2O2（4 小时）可诱导最大程度的细胞凋亡，但显著影响 Leydig 细胞活力。因此，在相同浓度下，进一步研究了凋亡标记基因 caspase-8、caspase-9、caspase-3 和多聚（腺嘌呤二核苷酸）核糖聚合酶的表达。用 20 nM 的 siRNA 孵育 caspase-8 或 caspase-9，可分别抑制其表达 55-60%和活性 50-55%。siRNA 的抑制效率与细胞的 post-H2O2 或 pre-H2O2 处理相当。类似地，Eugenia jambolana（100μg/ml）植物提取物也能有效抑制所有凋亡标记蛋白的过度表达。通过 siRNA 沉默 caspase 8 的表达而不是 caspase 9 的表达，可显著抑制 caspase 3 的活性，这意味着 H2O2 诱导的 Leydig 细胞凋亡主要通过外源性途径，随后扩展到其他途径。
