PDGF-BB对光老化皮肤成纤维细胞TIMPs及胶原蛋白的影响

The effect of PDGF-BB on TIMPs and collagen of skin fibroblasts after photoaging

目的探讨血小板衍生因子-BB(PDGF-BB)对光老化成纤维细胞(FBs)中胶原蛋白(COL)、基质金属蛋白酶(MMPs)、基质金属蛋白酶的组织抑制剂(TIMPs)的影响。方法利用cck-8试剂盒检测PDGF-BB对FBs增殖活性的影响;利用低剂量UVB重复照射成功建立FBs早衰模型;在末次造模结束24 h,利用Real-Time PCR技术检测MMPs的表达;末次造模结束48 h,利用West blot技术检测TIMPs和胶原的表达。结果在一定浓度范围(0～8 ng/ml)及处理时间(0～48 h)内,PDGF-BB对细胞有促进增殖作用,其中以1 ng/ml作用最佳。1 ng/ml PDGF-BB能缓解UVB诱导的TIMPs表达抑制(P<0.05),同时,能在蛋白水平上减缓UVB对FBsⅠ型及Ⅲ型胶原合成的抑制(P<0.05)。但对UVB诱导的MMPs的过表达无明显改善作用。结论 PDGF-BB虽然对MMPs无明显改善作用,但其能有效增加光老化FBs中TIMPs及Ⅰ型和Ⅲ型胶原的表达,调节光老化皮肤ECM合成/降解稳态,有望成为对抗光老化的新手段。

Objective To explore the effect of platelet-derived growth factor-BB(PDGF-BB) on collagen(COL), matrix metalloproteinases(MMPs), and tissue inhibitor of metalloproteinases(TIMPs) of skin fibroblasts(FBs) after photoaging. Methods The effect of PDGF-BB on FBs proliferation was detected by cell counting kit-8 assay. The FBs senescence model was successfully established by repeated irradiation of lowdose UVB; The influence of PDGF-BB on the expression of MMPs was evaluated through Real-Time PCR at 24 h after the last radiation. The synthesis of collagen and TIMPs was detected by Western Blot at 48 h after the last radiation. Results In a certain concentration range(0 to 8 ng/ml) and treatment period(0 to 48 h), PDGF-BB can promote cell proliferation; 1 ng/ml was the optimal. When the concentration of PDGF-BB is 1 ng/ml, it can alleviate the expression inhibition of TIMPs induced by UVB(P<0.05). At the same time, it can slowdown the inhibition of collagen synthesis induced by UVB in protein level(P<0.05). However, the overexpression of MMPs induced by UVB was not significantly improved. Conclusion Although PDGF-BB has no significant effect on MMPs, it can effectively increase the expression of TIMP and collagen in FBs and regulate the ECM synthesis/degradation homeostasis of photoaging skin. It is a promising way to ameliorate photoaging.