AIM: To examine the protective effect of estradiol on the cultured hepatocytes under oxidative stress.METHODS: Hepatocytes of rat were isolated by using perfusion method, and oxidative stress wes induced by a serum-fr...AIM: To examine the protective effect of estradiol on the cultured hepatocytes under oxidative stress.METHODS: Hepatocytes of rat were isolated by using perfusion method, and oxidative stress wes induced by a serum-free medium and FeNTA. MDA level was determined with TBA method. Cell damage was assessed by LDH assay. Apoptosis of hepatocytes was assessed with cytoflowmetric analysis. Expression of Bcl-xl in cultured hepatocytes was detected by Western blot.The radicalscavenging activity of estradiol was valued by its ability to scavenge the stable free radical of DDPH.RESULTS: Oxidative stress increased LDH (from 168 ± 25 x10-6 IU. cell 1 to 780 ± 62 x 10-6 IU. cell-1 ) and MDA(from 0.28 ±0.07 x 10-6 nmol. cell-1 to 1. 35 ± 0.12 × 10-6 nmol. cell-1 ) levelsin cultured hepatocyte, and estradiol inhibited both LDH andMDA production in a dose dependent manner. In thepresence of estradiol t0-6 mol. L-1, 107 mol. L-1 and 10-8 mol.L-1 ,the LDH levels are 410 ± 53 × 10-6 IU. cell-1 ( P ＜ 0.01 vsoxidative group), 530 ± 37 × 10-6 IU. cell-1 ( P ＜ 0. 01 vsoxidative group), 687 ± 42 x 10-6 IU. cell-1 ( P ＜ 0.05 vsoxidative group) respectively, and the MDA level are 0.71 ±0.12 x 10-6 nmol. cell-1 ( P ＜ 0.01 vs oxidative group), 0.97 ± 0.11 × 10-6 nmol. cell-1 ( P ＜ 0.01 vs oxidative group) and 1.27 ±0. 19 x 10-6 nmol. cell-1 respectively. Estradiol suppressedapoptosis of hepatocytes induced by oxidative stress,administration of estradiol (10-6 mol/ L)decreased theapoptotic rate of hepatocytes under oxidative stress from 18.6 ± 1.2% to 6.5 ± 2.5%, P ＜ 0.01. Bcl-xl expression wasrelated to the degree of liver cell damage due to oxidativestress, and estradiol showed a protective action.CONCLUSION: Estmdiol protects hepatocytes from oxidativedamage by means of its antioxidant activity.展开更多
文摘AIM: To examine the protective effect of estradiol on the cultured hepatocytes under oxidative stress.METHODS: Hepatocytes of rat were isolated by using perfusion method, and oxidative stress wes induced by a serum-free medium and FeNTA. MDA level was determined with TBA method. Cell damage was assessed by LDH assay. Apoptosis of hepatocytes was assessed with cytoflowmetric analysis. Expression of Bcl-xl in cultured hepatocytes was detected by Western blot.The radicalscavenging activity of estradiol was valued by its ability to scavenge the stable free radical of DDPH.RESULTS: Oxidative stress increased LDH (from 168 ± 25 x10-6 IU. cell 1 to 780 ± 62 x 10-6 IU. cell-1 ) and MDA(from 0.28 ±0.07 x 10-6 nmol. cell-1 to 1. 35 ± 0.12 × 10-6 nmol. cell-1 ) levelsin cultured hepatocyte, and estradiol inhibited both LDH andMDA production in a dose dependent manner. In thepresence of estradiol t0-6 mol. L-1, 107 mol. L-1 and 10-8 mol.L-1 ,the LDH levels are 410 ± 53 × 10-6 IU. cell-1 ( P ＜ 0.01 vsoxidative group), 530 ± 37 × 10-6 IU. cell-1 ( P ＜ 0. 01 vsoxidative group), 687 ± 42 x 10-6 IU. cell-1 ( P ＜ 0.05 vsoxidative group) respectively, and the MDA level are 0.71 ±0.12 x 10-6 nmol. cell-1 ( P ＜ 0.01 vs oxidative group), 0.97 ± 0.11 × 10-6 nmol. cell-1 ( P ＜ 0.01 vs oxidative group) and 1.27 ±0. 19 x 10-6 nmol. cell-1 respectively. Estradiol suppressedapoptosis of hepatocytes induced by oxidative stress,administration of estradiol (10-6 mol/ L)decreased theapoptotic rate of hepatocytes under oxidative stress from 18.6 ± 1.2% to 6.5 ± 2.5%, P ＜ 0.01. Bcl-xl expression wasrelated to the degree of liver cell damage due to oxidativestress, and estradiol showed a protective action.CONCLUSION: Estmdiol protects hepatocytes from oxidativedamage by means of its antioxidant activity.
