银杏内酯B对H2O2诱导人视网膜色素上皮细胞氧化损伤的保护作用

目的　探讨银杏内酯Ｂ对体外培养的人视网膜色素上皮（ｒｅｔｉｎａｌｐｉｇｍｅｎｔｅｐｉｔｈｅｌｉａｌ，ＲＰＥ）细胞氧化损伤的保护作用及可能机制。方法　ＲＰＥ细胞传代培养２４ｈ后，随机分为阴性对照组、氧化损伤组、银杏内酯Ｂ低浓度组（１ｍｏｌ?Ｌ－１）和银杏内酯Ｂ高浓度组（１０ｍｏｌ?Ｌ－１），银杏内酯Ｂ预处理２４ｈ后，加入１００μｍｏｌ?Ｌ－１Ｈ２Ｏ２继续孵育１２ｈ，ＭＴＴ比色法检测细胞活力，流式细胞仪检测细胞凋亡率，Ｈｏｃｈｅｓｔ３３２５８染色观察凋亡细胞形态，比色法检测凋亡相关因子Ｃａｓｐａｓｅ-３及Ｃａｓｐａｓｅ-９的表达。结果　银杏内酯Ｂ能明显抑制Ｈ２Ｏ２诱导的ＲＰＥ细胞活力的下降，ＭＴＴ结果显示银杏内酯Ｂ低浓度组和银杏内酯Ｂ高浓度组细胞活性分别为（５３．３７±２．５３）％和（６９．５７±３．１７）％，与氧化损伤组的（３１．３３±２．４１）％比较，差异均具有统计学意义（均为Ｐ＜０．０５）；流式细胞计数结果显示银杏内酯Ｂ低浓度组和银杏内酯Ｂ高浓度组细胞凋亡率分别下降至（２７．５３±３．３４）％和（１３．３０±２．２５）％，与氧化损伤组的（４８．１３±２．６８）％比较，差异均具有统计学意义（均为Ｐ＜０．０１）。此外，银杏内酯Ｂ还可以减少Ｈ２Ｏ２所致ＲＰＥ细胞内Ｃａｓｐａｓｅ-３及Ｃａｓｐａｓｅ-９的表达。结论　银杏内酯Ｂ通过抑制Ｃａｓｐａｓｅ-３及Ｃａｓｐａｓｅ-９的表达有效抑制了Ｈ２Ｏ２对ＲＰＥ细胞的损伤，从而为其用于治疗ＲＰＥ细胞损伤提供可靠的实验依据。

Protective effects of ginkgolide B on H2O2-induced oxidative damage in RPE cells

Objective To discuss the protective effects and possible mechanisms of ginkgolide B on oxidative damage of human retinal pigment epithelial ( RPE) cells cultured in vitro. Methods RPE cells were cultured and passaged for 24 hours . and randomly divided into four groups: Negative control group, oxidative damage group, ginkgolide B low dose ( I mol . L-’) group and ginkgolide B high dose ( 10 mol . L’1 ) group ,pre-treated with ginkgolide B for 24 hours , and then treated with 100 Vmol . L -l H202 for 12 hours, cell viability were tested by MTT colorimetric detection, apoptotic rate was detected by flow cytometry, apoptotic cell morphology was observed by hochest33258 staining, expression of apoptosis-related factors Caspase-3 and Caspase-9 were tested by colorimetric detection. Results Ginkgolide B could effective inhibit H202-induced cells viability decrease in RPE cells. MTT assay results showed that cell viability in ginkgolide B low dose group and ginkgolide B high dose group were ( 53. 37 + 2. 53 ) % and( 69. 57 + 3. 17 ) % , there were statistically sigruficant differences compared with oxidative damage group ( 31. 33 + 2. 41 ) % ( all P < 0. 05 ) ; Flow cell count results showed that cell apoptotic rates in ginkgolide B low dose group and ginkgolide B high dose group decreased to ( 27. 53 +3. 34) % and ( 13. 30 +2. 25 ) % ,there were statistically significant differences compared with oxidative damage group (48. 13 +2. 68 ) % ( all P < 0. 01 ). Besides, ginkgolide B could decrease the expression of Caspase-3 and Caspase-9 in H2 02-induced oxidative damage of RPE cells. Conclusion Ginkgolide B can effectively inhibit H202-induced RPE cells damage by inhibiting the expression of Caspase-3 and Caspase-9 ,which provide reliable experimental basis for the treatment of injuries in RPE cells.