| 王 洁,何振兴,赵菊花,李 达,李 燃,杨 羽.绞股蓝总皂苷对光老化人皮肤成纤维细胞凋亡及Caspase-3信号通路的影响[J].,2017,17(19):3632-3635 |
| 绞股蓝总皂苷对光老化人皮肤成纤维细胞凋亡及Caspase-3信号通路的影响 |
| The Effect of Gypenosides on Cell Apoptosis and Caspase-3 Signaling Pathway in Human Skin Fibroblasts |
| 投稿时间：2017-03-16 修订日期：2017-04-05 |
| DOI：10.13241/j.cnki.pmb.2017.19.008 |
| 中文关键词: 绞股蓝总皂苷 光老化 皮肤成纤维细胞 凋亡 Caspase-3 信号通路 |
| 英文关键词: Gypenosides Photo-aging Human skin fibroblasts cell Apoptosis Caspase-3 Signaling pathway |
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| 中文摘要: |
| 摘要 目的：探讨绞股蓝总皂苷（Gyp）对光老化人皮肤成纤维细胞（HSF细胞）凋亡以及Caspase-3信号通路的影响。方法：分别以80、160、320 mg/d剂量的Gyp生理盐水溶液灌胃大鼠7d后取血并分离血清，长波紫外线（UVA）照射方法（照射剂量36 J/cm3）构建光老化HSF细胞模型以得到低剂量组、中剂量组、高剂量组，同时以空白对照组（未照射细胞）、UVA模型组、正常组为对照。UVA诱导的细胞活性氧表达采用二氯荧光素（DCF）法测定，细胞凋亡情况采用TUNEL法测定，HSF细胞活性采用四甲基偶氮唑盐微量酶反应比色法（MTT法）测定，Bax、Bcl-2、Caspase-3基因和蛋白表达分别采用反转录-聚合酶链反应（RT-PCR）和West- ern-blotting方法进行测定。结果：与空白对照组比较，其余5组的OD值、HSF细胞凋亡数、活性氧（平均荧光强度）、活性氧水平、Bax、Bcl-2、Caspase-3 mRNA及蛋白表达水平差异具有统计学意义（P<0.05）；与UVA模型组和正常组比较，低、中、高剂量组OD值、HSF细胞凋亡数、活性氧（平均荧光强度）、活性氧水平、Bax、Bcl-2、Caspase-3 mRNA及蛋白表达水平差异具有统计学意义（P<0.05）；低、中、高剂量组随着剂量增加OD值、Bcl-2mRNA和蛋白表达水平逐渐升高，细胞凋亡数、活性氧（平均荧光强度）、活性氧水平、Bax、Caspase-3 mRNA和蛋白表达水平逐渐降低（P<0.05）。结论：Gyp通过抑制细胞内活性氧的产生以及Bax的表达，以及激活Bcl-2、Caspase-3信号通路而逆转UVA诱导的HSF细胞凋亡，进而延缓HSF细胞的光老化现象。 |
| 英文摘要: |
| ABSTRACT Objective: To explore the influence of gypenosides (Gyp) on cell apoptosis and Caspase-3 signaling pathway in pho- to-aging human skin fibroblasts(HSF cell). Methods: Blood samples of rats gavaged to Gyp saline solution with 80, 160, 320 mg/d re- spectively after 7 days were taken, and the serum was isolated. The photo-aging HSF cell model were established by the long-wave ultra- violet(UVA)(irradiation dose 36 J/cm3), which were divided into Low-dose group, Middle-dose group and High-dose group,and the blank control group(without irradiated cells), UVA model group,Normal group were selected as control group. The activity of reactive oxygen species induced by UVA was determined by the dichlorofluorescein (DCF) method, the cell apoptosis was measured by the TUNEL method, the activity of HSF cells was determined by the MTT method, the gene an protein expression of Bax, Bcl-2, Caspase-3 were de- tected by the reverse transcription-polymerase chain reaction (RT-PCR) and Western-blotting respectively. Results: The OD value, HSF cell apoptosis number, the reactive oxygen species(average fluorescence intensity), reactive oxygen species levels, mRNA and protein ex- pression of Bax, Bcl-2 and Caspase-3 in the other 5 groups compared with blank control group, the differences were statistically signifi- cant (P<0.05); The OD value, HSF cell apoptosis number, the reactive oxygen species(average fluorescence intensity),reactive oxygen species levels, mRNA and protein expression of Bax, Bcl-2 and Caspase-3 in Low, Middle, High-dose group compared with UVA model group and Normal group, the differences were statistically significant (P<0.05); Low, medium and high -dose group increased with the dose of OD value, mRNA and protein expression of Bcl-2 increased gradually, HSF cell apoptosis number, the reactive oxygen species(average fluorescence intensity), reactive oxygen species levels, mRNA and protein expression of Bax and Caspase-3 decreased gradually(P<0.05). Conclusion: Gyp can delay the aging of HSF cell by reversing HSF cell apoptosis induced by UVA, the possible mechanism is inhibition of the production of intracellular reactive oxygen species and expression of Bax, and activation of Bcl-2, Caspase-3 signaling pathway. |
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