绞股蓝总皂苷对光老化人皮肤成纤维细胞凋亡的影响

Effects of gypenosides on the apoptosis of photoaging human dermal fibroblasts

目的:观察绞股蓝总皂苷(Gyp)对光老化人皮肤成纤维细胞株HSF凋亡的影响。方法:分别按80、160和320 mg/d的剂量灌胃大鼠Gyp生理盐水溶液,7 d后取血,分离血清备用。采用长波紫外线(UVA)照射方法建立光老化HSF细胞模型,照射剂量36 J/cm2,然后用制备的低、中、高剂量Gyp含药血清进行干预培养。12 h后,采用TUNEL法检测细胞凋亡情况,采用RT-PCR和Western blot法检测细胞中Bax、Bcl-2 mRNA和蛋白的表达。以未照射细胞(空白对照组)、模型细胞和正常血清干预的模型细胞(正常血清组)作对照。结果:6组凋亡细胞数,细胞中Bcl-2、Bax mRNA和蛋白的表达水平差异均有统计学意义(F=474.201,351.094、334.907和403.610、162.766,P<0.001)。与空白对照组比较,UVA模型组和正常血清组细胞凋亡增强,Bcl-2 mRNA和蛋白表达降低,同时Bax mRNA和蛋白表达升高(P<0.05)。与UVA模型组和正常血清组比较,低、中、高剂量Gyp组细胞凋亡减弱,细胞中Bcl-2 mRNA和蛋白表达升高,Bax mRNA和蛋白表达降低(P<0.05),高剂量组效果更显著(P<0.05)。结论:Gyp可以逆转UVA诱导的人皮肤成纤维细胞的凋亡。

Aim：To investigate the effect of gypenosides （ Gyp） on the apoptosis of photoaging human dermal fibro-blasts（HSF cells） induced by ultraviolet A（UVA） radiation.Methods：Rats were intragastricly administrated Gyp saline solution at doses of 80, 160 and 320 mg/d, respectively.7 d later, the serum was isolated.HSF cells were irradiated by UVA at a dose of 35 mJ/cm2 to establish photoaging cell model ,then these cells were cultured by low-,median-and high-dose Gyp medicated serum（Gyp groups）.12 h later, apoptosis was detected by TUNEL , the expressions of Bax and Bcl-2 mRNA and protein in cells were detected using RT-PCR and Western blot ,respectively .The cells without irradiation （ blank control group）, model cells without intervention（model group） and with normal serum intervention （normal serum group） were the controls.Results： There were significant differences in apoptosis cell number ,expressions of Bcl-2 mRNA,Bax mRNA,Bcl-2 and Bax protein in cells among the six groups （F=474.201,351.094,334.907 and 403.610,162.766,P〈0.001）.Compared with the blank control group , apoptosis of model group and normal serum group increased , and the ex-pressions of Bcl-2 mRNA and protein decreased,while those of Bax increased significantly （P〈0.05）.Compared with the model group and normal serum group , apoptosis of Gyp groups decreased ,and the expressions of Bcl-2 mRNA and protein increased,while those of Bax decreased significantly （P〈0.05）,especially the high-dose Gyp group（P〈0.05）.Conclu-sion：Gyp can protect HSF cells from UVA-induced apoptosis .