自噬在富氢液减轻UVB致皮肤成纤维细胞损伤中的作用

Hydrogen Alleviated Cellular Injury in Human Fibroblast Induced by UVB Irradiation Via Activating Autophagy

目的探讨自噬对富氢液减轻UVB照射导致的人皮肤成纤维细胞氧化损伤的调控作用。方法取健康男童包皮环切术后包皮,进行人真皮成纤维细胞的原代培养,将细胞分组:正常对照组(Con组);氢气对照组(Con+H_2);UVB照射组(UV组);UVB照射+富氢培养液组(UV+H_2组);UVB照射+富氢培养液+自噬抑制剂3-MA组(UV+H_2+3-MA组),各组培养6h、12h和24h,电镜观察自噬体和LC3的表达变化,根据上述结果,各组在UVB照射后培养6h后,进行相MTT、LDH、ROS、MDA和自噬蛋白Beclin-1、ATG5和P62的检测。结果与Con组和Con+H_2组比较,UV组MTT值和P62表达减少(P<0.05),自噬体、LDH、ROS、MDA、LC3Ⅱ/Ⅰ、Beclin-1和ATG5表达增加(P<0.05);与UV组比较,UV+H_2组自噬体、MTT、LC3Ⅱ/Ⅰ、Beclin-1和ATG5表达增加(P<0.05),LDH、ROS、MDA和P62表达减少(P<0.05);与UV+H_2组比较,UV+H_2+3-MA组自噬体、MTT、LC3Ⅱ/Ⅰ、Beclin-1和ATG5表达减少(P<0.05),LDH、ROS、MDA和P62表达增加(P<0.05)。结论富氢液通过激活自噬途径减轻UVB照射导致的人皮肤成纤维细胞损伤。

Objective To investigate the regulatory role of autophagy in alleviating cellular oxi- dative injury by hydrogen in human fibroblast that induced by UVB irradia- tion. Methods Dermal fibroblasts were isolated from the circumcised foreskin of healthy boys to manage the primary culture, which were then divided into five groups： control group （ Con group）, control ＋ H2 group （ Con ＋ H2 group）, UVB irradiation group （ UV group）, UVB irradiation ＋ H2 group （ UV ＋ H2 group）, and UVB irradia- tion ＋ H2 ＋ autophagy inhibitor 3-MA group （ UV ＋ H2 ＋ 3-MA group）. The autopha- gosome and LC3 expression were detected at 6h, 12h and 24h after UV irradiation in different groups, and on the basis of the results above, MTY, LDH, ROS, MDA au- tophagosome and autophagy proteins Beclin-1, ATG5 and P62 were detected at 6h af- ter UV irradiation in different groups. Results Compared with Con group and Con ＋ H2 group, MTT and P62 reduced, autophagosome, ROS, MDA, LC3 Ⅱ/Ⅰ , Beclin- 1 and ATG5 were increased in UV group （P 〈 0.05 ） ; compared with UV group, au- tophagosome, MTY, LC3 Ⅱ/Ⅰ, Beclin-1 and ATG5 increased, LDH, ROS, MDA and P62 reduced in UV ＋ H2 group （P 〈 0. 05） ; compared with UV ＋ H2 group, au- tophagosome, MTY, LC3 Ⅱ/ Ⅰ, Beclin-1 and ATG5 were increased（ P 〈 0. 05 ） , and LDH, ROS, MDA and P62 were elevated in UV ＋ H2 ＋ 3-MA group （P 〈 0. 05）. Conclusion Hydrogen-rich culture alleviated the cellular injury via activating auto- phagy in human fibroblast induced by UVB irradiation.