蒿秦化斑方对紫外线B诱导的HaCaT细胞光损伤的影响

目的观察蒿秦化斑方对紫外线B(ultraviolet-B,UVB)诱导的HaCaT细胞光损伤的保护作用并探讨其可能机制。方法建立300 mJ/cm 2 UVB辐射损伤HaCaT细胞的病理模型,使用蒿秦化斑方干预,实验分为空白对照组、UVB模型组及蒿秦化斑方高、中、低剂量组。使用MTT法测定蒿秦化斑方的安全性及HaCaT细胞的增殖;吸光度法测定HaCaT细胞中超氧化物歧化酶(superoxide dismutase,SOD)、过氧化氢酶(catalase,CAT)的活性和丙二醛(malondialdehyde,MDA)的含量;ELISA法检测HaCaT细胞细胞间粘附分子-1(intercellular adhesion molecule 1,ICAM-1)、血管细胞间黏附分子-1(vascular cell adhesion molecule 1,VCAM-1)、内皮细胞选择素(endothelium-selectin,E-selectin)、白细胞介素-10(interleukin-10,IL-10)、转化生长因子β(transforming growth facor-β,TGF-β)的含量。结果与空白对照组比较,UVB模型组细胞增殖能力降低(P<0.05),与UVB模型组比较,蒿秦化斑方高、中、低剂量组细胞增殖能力均增强(P<0.05);与空白对照组比较,UVB模型组SOD、CAT活性降低(P<0.05),MDA含量增加(P<0.05);与UVB模型组比较,蒿秦化斑方高剂量组SOD、CAT的活性增强(P<0.05),MDA的含量降低(P<0.05)。与空白对照组比较,UVB模型组ICAM-1、VCAM-1、E-selectin、IL-10含量均增加(P<0.05),TGF-β含量表达降低(P<0.05);与UVB模型组比较,蒿秦化斑方高剂量组ICAM-1、VCAM-1、E-selectin、IL-10的含量均降低(P<0.05),TGF-β含量升高(P<0.05)。结论高剂量蒿秦化斑方能够通过增强抗氧化酶SOD、CAT的活性及减少脂质过氧化物MDA的产生抑制UVB诱导的HaCaT细胞氧化损伤,通过降低HaCaT细胞中炎症相关因子ICAM-1、VCAM-1、E-selectin、IL-10的表达抑制UVB诱导的HaCaT细胞炎症反应,通过细胞增殖和上调TGF-β的表达修复HaCaT细胞屏障功能,从而对UVB诱导的HaCaT细胞光损伤起到保护作用。

Effects of Haoqin Huaban formula on photic injury of HaCaT cells induced by ultraviolet-B

Objective To observe the protective effect of Haoqin Huaban formula on photic injury of HaCaT cells induced by ultraviolet-B(UVB),and explore its possible mechanism.Methods A pathological model of HaCaT cells injured by 300 mJ/cm 2 UVB radiation was established.The intervention with Haoqin Huaban formula was carried out,and the experiment was divided into blank group,UVB model group,and high,medium and low dose of Haoqin Huaban formula groups.The safety of Haoqin Huaban formula and HaCaT cells’proliferation were detected by MTT method.The activities of superoxide dismutase(SOD)and catalase(CAT)and the content of malonaldehyde(MDA)in HaCaT cells were detected by absorbance method.The intercellular adhesion molecule-1(ICAM-1),vascular cell adhesion molecule-1(VCAM-1),endothelium-selectin(E-selectin),interleukin-10(IL-10)and transforming growth factor-β(TGF-β)of HaCaT cells were detected by ELISA.Results Compared with the blank group,the cell proliferation ability in the UVB model group were weakened(P<0.05).Compared with the UVB model group,the cell proliferation ability in the high,medium,and low dose groups of Haoqin Huaban formula all were strengthened(P<0.05).Compared with the blank group,the activities of SOD and CAT in the UVB model group decreased(P<0.05),and the content of MDA increased(P<0.05).Compared with the UVB model group,the activities of SOD and CAT in the high dose of Haoqin huaban formula group increased(P<0.05),and the content of MDA decreased(P<0.05).Compared with the blank group,the contents of ICAM-1,VCAM-1,E-selectin and IL-10 in the UVB model group increased(P<0.05),and the content of TGF-βdecreased(P<0.05).Compared with the UVB model group,the contents of ICAM-1,VCAM-1,E-selectin and IL-10 in the high dose of Haoqin Huaban formula group decreased(P<0.05),and the content of TGF-βincreased(P<0.05).Conclusion High dose of Haoqin Huaban formula group can inhibit the oxidative damage of HaCaT cells induced by UVB,through enhancing the activities of antioxidant enzymes SOD and CAT and reducing the production of lipid peroxide MDA.It can inhibit HaCaT cells’inflammatory response by reducing the expression of inflammation related factors,such as ICAM-1,VCAM-1,E-selectin,and IL-10.It can repair the barrier function of HaCaT cells by cell proliferation and up-regulating the expression of TGF-β.Thus,Haoqin Huaban formula can prodect HaCaT cells from photic injury induced by UVB.