精子顶体蛋白酶是精子在女性生殖道内运行及穿透卵透明带与卵子结合而完成受精的特异性主酶之一。检测精子顶体蛋白酶活性(APA)对正确评价精子的受精能力及探讨顶体酶抑制剂的机理,筛选及开发新型避孕药都有明确的意义。本文介绍经本实...精子顶体蛋白酶是精子在女性生殖道内运行及穿透卵透明带与卵子结合而完成受精的特异性主酶之一。检测精子顶体蛋白酶活性(APA)对正确评价精子的受精能力及探讨顶体酶抑制剂的机理,筛选及开发新型避孕药都有明确的意义。本文介绍经本实验室改良的明胶底物玻片法检测人精子顶体蛋白酶活性,以精子顶体溶解明胶基质所显示的平均光轮直径(MHD)作为 APA 的指标,测得正常功能精子 APA 的 MHD 为29.96μm。证实该法能最大限度地显示 APA的光轮直径,操作简便、实用,可有效地用于临检。同时以该法测到顶体酶抑制剂 TLCK 的浓度梯度效应,验证了该法的可靠性。展开更多
Hydrogen peroxide (H2O2) induces the hypertrophy in cultured H9c2 cardiomyocytes and cell death in glutathione (GSH)-depleted H9c2 cells. In the present study, we observed that pretreatment with a serine protease inhi...Hydrogen peroxide (H2O2) induces the hypertrophy in cultured H9c2 cardiomyocytes and cell death in glutathione (GSH)-depleted H9c2 cells. In the present study, we observed that pretreatment with a serine protease inhibitor, N-a-tosyl-L-lysine chloromethyl ketone (TLCK), significantly prevented the H2O2-induced cell damages in GSH-depleted H9c2 cells in a concentration-dependent manner. The phase contrast microscopy revealed that although the exposure of the GSH-depleted H9c2 cells to H2O2 resulted in a globular shape of the cells, TLCK prevented the occurrence of H2O2-induced morphological changes. TLCK also inhibited the generation of reactive oxygen species in the cells after addition of H2O2, suggesting that the antioxidant action of TLCK is involved in the protection against the cell damages by H2O2. Application of TLCK after ~30 min of exposure to H2O2 could significantly protect the cells from cell damages. The other serine protease inhibitors that were tested could not prevent the cell damages in GSH- depleted H9c2 cells. Pretreatment with an inhibitor of nuclear factor-kB translocation into the nucleus and a proteasome inhibitor did not prevent the cell damages in GSH-depleted H9c2 cells. An inhibitor of p53 significantly prevented the cell damages in GSH-depleted H9c2 cells. These results suggest that antioxidative action of TLCK prevents the death of GSH-depleted H9c2 cardiomyocytes induced by H2O2.展开更多
文摘精子顶体蛋白酶是精子在女性生殖道内运行及穿透卵透明带与卵子结合而完成受精的特异性主酶之一。检测精子顶体蛋白酶活性(APA)对正确评价精子的受精能力及探讨顶体酶抑制剂的机理,筛选及开发新型避孕药都有明确的意义。本文介绍经本实验室改良的明胶底物玻片法检测人精子顶体蛋白酶活性,以精子顶体溶解明胶基质所显示的平均光轮直径(MHD)作为 APA 的指标,测得正常功能精子 APA 的 MHD 为29.96μm。证实该法能最大限度地显示 APA的光轮直径,操作简便、实用,可有效地用于临检。同时以该法测到顶体酶抑制剂 TLCK 的浓度梯度效应,验证了该法的可靠性。
文摘Hydrogen peroxide (H2O2) induces the hypertrophy in cultured H9c2 cardiomyocytes and cell death in glutathione (GSH)-depleted H9c2 cells. In the present study, we observed that pretreatment with a serine protease inhibitor, N-a-tosyl-L-lysine chloromethyl ketone (TLCK), significantly prevented the H2O2-induced cell damages in GSH-depleted H9c2 cells in a concentration-dependent manner. The phase contrast microscopy revealed that although the exposure of the GSH-depleted H9c2 cells to H2O2 resulted in a globular shape of the cells, TLCK prevented the occurrence of H2O2-induced morphological changes. TLCK also inhibited the generation of reactive oxygen species in the cells after addition of H2O2, suggesting that the antioxidant action of TLCK is involved in the protection against the cell damages by H2O2. Application of TLCK after ~30 min of exposure to H2O2 could significantly protect the cells from cell damages. The other serine protease inhibitors that were tested could not prevent the cell damages in GSH- depleted H9c2 cells. Pretreatment with an inhibitor of nuclear factor-kB translocation into the nucleus and a proteasome inhibitor did not prevent the cell damages in GSH-depleted H9c2 cells. An inhibitor of p53 significantly prevented the cell damages in GSH-depleted H9c2 cells. These results suggest that antioxidative action of TLCK prevents the death of GSH-depleted H9c2 cardiomyocytes induced by H2O2.
