论文部分内容阅读
目的:研究京尼平苷对光老化HaCaT细胞的保护作用及其机制。方法:取对数生长期的HaCaT细胞,调整其密度为1×106个/mL接种于6孔板、5×104个/mL接种于96孔板,用总剂量64 mJ·cm-2(照射强度0.61 mW·cm-2×照射时间105 s)的紫外线B(UVB)照射细胞建立光老化模型,以5×10-5,5×10-6,5×10-7mol·L-1的京尼平苷处理光老化细胞24 h,分别检测细胞中超氧化物歧化酶(SOD)活性、谷胱甘肽过氧化物酶(GSH-Px)活性、丙二醛(MDA)含量,RT-PCR法检测细胞中P38、肿瘤坏死因子α(TNF-α)和白介素6(IL-6)mRNA的表达量,酶联免疫法检测细胞上清液中TNF-α和IL-6的含量。结果:UVB照射HaCaT细胞后,SOD活性、GSH-Px活性明显降低,MDA含量、p38 mRNA表达量、TNF-α和IL-6 mRNA及蛋白的表达量明显升高(P<0.01);5×10-5,5×10-6mol·L-1京尼平苷处理细胞后,SOD活性升高为(34.72±2.03),(38.96±3.56)U·mg-1;GSH-Px活性升高为(28.11±2.76),(40.02±3.63)U·mg-1;MDA含量下降为(4.82±0.42),(4.60±0.41)nmol·mg-1,p38 mRNA的相对含量下降为(0.97±0.09),(0.94±0.09);TNF-αmRNA的相对含量下降为0.57±0.05,0.53±0.05;IL-6 mRNA的相对含量下降为0.65±0.06,0.64±0.06;TNF-α蛋白的表达量降低为(34.05±2.43),(25.55±1.84)ng·L-1,IL-6蛋白的表达量降低为(28.51±1.95),(19.32±1.55)ng·L-1,与模型对照组相比,差异具有统计学意义(P<0.05,P<0.01)。结论:京尼平苷能抵抗UVB对HaCaT细胞的损伤,其机制可能与抑制氧化损伤,调控p38信号通路,调节TNF-α和IL-6的表达有关。
Objective: To study the protective effect and mechanism of geniposide on photodamaged HaCaT cells. Methods: The logarithmic growth phase HaCaT cells were seeded in a 6-well plate at a density of 1 × 106 cells / mL and seeded into 96-well plates at 5 × 104 cells / mL for a total dose of 64 mJ · cm-2 (UVB) irradiated cells with intensity of 0.61 mW · cm-2 × irradiation time of 105 s) were used to establish the photo-aging model. The photo-aging model was established with 5 × 10-5, 5 × 10-6, 5 × 10-7 mol·L-1 The cells were treated with nipagin for 24 h. The activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) were detected by RT-PCR The expression of P38, TNF-α and IL-6 mRNA were detected by enzyme-linked immunosorbent assay (ELISA). The contents of TNF-α and IL-6 in the supernatant were detected by ELISA. Results: The activity of SOD and the activity of GSH-Px in UVB-irradiated HaCaT cells were significantly decreased. The contents of MDA, p38 mRNA, TNF-α and IL-6 mRNA and protein were significantly increased (P <0.01) The activity of SOD increased to (34.72 ± 2.03) and (38.96 ± 3.56) U · mg-1 after treatment with 10-5 and 5 × 10-6mol·L-1 of geniposid. The GSH-Px activity increased (28.11 ± 2.76) and (40.02 ± 3.63) U · mg-1, respectively. The content of MDA decreased (4.82 ± 0.42) and (4.60 ± 0.41) nmol · mg-1 and the relative content of p38 mRNA decreased to (0.97 ± 0.09) , (0.94 ± 0.09). The relative content of TNF-αmRNA decreased to 0.57 ± 0.05 and 0.53 ± 0.05 respectively. The relative content of IL-6 mRNA decreased to 0.65 ± 0.06 and 0.64 ± 0.06 respectively. The expression of TNF- 34.05 ± 2.43 and 25.55 ± 1.84 ng · L-1, respectively, and the expression of IL-6 protein decreased by (28.51 ± 1.95) and (19.32 ± 1.55) ng · L-1, respectively. Compared with the model control group, With statistical significance (P <0.05, P <0.01). Conclusion: Geniposide can resist the damage of HaCaT cells induced by UVB, which may be related to the inhibition of oxidative damage, the regulation of p38 signaling pathway and the regulation of TNF-α and IL-6 expression.