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目的:探讨桃叶珊瑚苷对紫外线B诱导的HaCaT细胞凋亡相关蛋白Bax、Bcl-2、Caspase-3表达的影响,进一步阐明桃叶珊瑚苷抗光老化的作用机制。方法:将指数生长期HaCaT细胞随机分为正常对照组、模型对照组、10~(-7)、10~(-6)、10~(-5) mol·L~(-1) AU处理组,采用64 m J·cm~(-2)的UVB照射建立细胞光损伤模型,以终浓度10~(-7)、10~(-6)、10~(-5)mol·L~(-1)AU处理光损伤细胞,试剂盒检测ROS含量、Caspase-3活性,流式细胞术检测细胞凋亡率,Western-Blot检测Bcl-2、Bax蛋白的表达量。结果:UV照射的HaCaT细胞ROS含量、细胞凋亡率、Caspase-3活性、Bcl-2、Bax蛋白表达量均升高(P<0.01),Bcl-2/Bax值降低(P<0.01);不同浓度桃叶珊瑚苷处理后,ROS含量、Caspase-3活性、Bax蛋白表达量降低,Bcl-2蛋白表达量、Bcl-2/Bax值升高,10~(-6)、10~(-5) mol·L~(-1)桃叶珊瑚苷组与模型对照组相比,差异具有统计学意义(P<0.05,0.01)。结论:桃叶珊瑚苷通过清除ROS,下调Bax、Caspase-3表达,上调Bcl-2表达,抑制细胞凋亡,保护受损的HaCaT细胞,拮抗光老化。
OBJECTIVE: To investigate the effect of aucubin on the expression of Bax, Bcl-2 and Caspase-3 in HaCaT cells induced by ultraviolet B and to elucidate the mechanism of aucubin against photoaging. Methods: The exponential growth phase HaCaT cells were randomly divided into normal control group, model control group, 10 ~ (-7), 10 ~ (-6), 10 ~ (-5) mol·L ~ The model of photodamage was established by UVB irradiation of 64 mJ · cm ~ (-2) at the final concentration of 10 -7, 10 -6, 10 -5 mol·L -1, 1) AU treated photodamaged cells. The activity of ROS and Caspase-3 were detected by kit. The apoptosis rate was detected by flow cytometry. The expression of Bcl-2 and Bax protein was detected by Western-Blot. Results: The ROS content, apoptosis rate, Caspase - 3 activity, Bcl - 2 and Bax protein expression increased (P <0.01) and Bcl-2 / Bax decreased in UV irradiated HaCaT cells (P <0.01). The contents of ROS, Caspase-3, Bax, Bcl-2 protein and Bcl-2 / Bax were decreased after treatment with aucubin at different concentrations for 10 ~ (-6), 10 ~ 5) mol·L -1 aucubin group compared with the model control group, the difference was statistically significant (P <0.05,0.01). CONCLUSION: Aucubin protects damaged HaCaT cells against ROS through down-regulating ROS, down-regulating Bax and Caspase-3 expression, up-regulating Bcl-2 expression and inhibiting apoptosis.