五倍子水提物对UVB诱导光老化成纤维细胞的保护作用

Protective Effect of Galla Chinensis Extract on UVB-Induced Photoaging in Human Skin Fibroblast

目的探究五倍子水提物(galla chinensis extract,GCE)对中波紫外线(UVB)诱导的人皮肤成纤维细胞(HSF)光老化的保护作用及机制。方法通过自由基清除实验、弹性蛋白酶抑制实验评价五倍子水提物的抗衰老能力;建立UVB诱导的细胞光老化模型,给予五倍子水提物干预,将细胞分为对照组、模型组(UVB)和实验组(GCE)。通过CCK-8法检测细胞存活率,β-半乳糖苷酶染色法检测细胞衰老水平,流式细胞仪检测活性氧(ROS),试剂盒检测细胞中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、丙二醛(MDA)的活性;通过酶联免疫吸附试验(ELISA)检测基质金属蛋白酶-1(MMP-1)、白细胞介素-6(IL-6)、Ⅰ型胶原蛋白(COL-1)含量。蛋白免疫印迹法(Western blot)检测相关蛋白表达水平。结果五倍子水提物具有清除自由基、抑制弹性蛋白酶活性、显著提高UVB诱导的HSF存活率、COL-1含量、抗氧化物酶(CAT、SOD)活性等作用,与对照组及UVB组相比差异有统计学意义(P<0.05)。同时,五倍子水提物还可以显著抑制UVB诱导的HSF衰老,ROS、IL-6、MMP-1和MDA生成以及抑制丝裂原活化蛋白激酶(MAPK)信号通路中p-ERK1/2、p-JNK及p-p38蛋白的表达水平,与对照组及UVB组相比差异有统计学意义(P<0.05)。结论五倍子水提物可通过提高抗氧化酶活力以及抑制MAPK信号通路的激活,发挥抗光老化作用。

Objective To investigate the protective effect and mechanism of galla chinensis extract(GCE)onhumanskinfibroblasts(HSF)photoaginginduced by medium wavelength ultraviolet(UVB)light.MethodsTheanti-agingability oftheGCE was evaluated by free radical scavenging assay and elastase inhibition assay;a UVB-induced cellular photoaging model was established,and the GCE was given to intervene.The cells were divided into control group,model group(UVB),and experimental group(GCE).The cell viability was detected by CCK-8 assay,the senescence level was detected byβ-galactosidase staining assay,the reactive oxygen species(ROS)level was detected by cellflow cytometry,the activities of superoxide dismutase(SOD),catalase(CAT),andmalondialdehyde(MDA)weredetected using the kit;the levels of matrix metalloproteinase-1(MMP-1),interleukin-6(IL-6),and type I collagen(COL-1)were detected by enzyme-linked immunosorbent assay(ELISA).The expression levels of related proteins were detected by protein immunoblotting(Western blot).Results CCE had the positive effect of scavenging free radicals,inhibiting elastase activity,increasing the survival rate of UVB-induced HSF cells,the content of COL-1,and the activities of antioxidant enzymes(CAT,SOD).There were statistically significant differences compared with the control group and the UVB group(P<0.05).At the same time,GCE could also inhibite UVB-induced senescence of HSF cells,the production of ROS,IL-6,MMP-1,MDA,and the p-ERK1/2,p-JNK,p-p38 protein expression levels in the mitogen-activated protein kinase(MAPK)signalingpathway.Therewerestatistically significant differences compared with the control group and the UVB group(P<0.05).Conclusion The GCE exerts anti-photoaging effects by increasing the activity of antioxidant enzymes and inhibiting the activation of the MAPK signaling pathway.