目的建立视网膜光感受器细胞外基质(Interphotoreceptor matrix,IPM)蛋白质sialoprotein associated with cones and rods(SPACR)分离和纯化方法。方法分离视网膜IPM,提取蛋白质组份,通过离子交换、凝胶过滤层析等技术分离纯化SPACR,并通过双向电泳及western blot技术验证。结果通过双向电泳技术及western blot结果显示分离纯化后仅有等电点在5-5.5,分子量约在150 kDa的单一条带和SPACR抗体反应,表明得到高纯度的SPACR。结论通过本文所述方法可以得到纯度较高的SPACR,为将来进一步对SPACR活性及结构研究奠定了基础。
To investigate the effects of topically applied 17β-estradiol on the expression of extracellular matrix proteins in aged human skin, 17β-estradiol (0.01%) and its vehicle (70%propylene glycol, 30%ethanol) were applied to aged (68-82 y, eight females and five males) human buttock skin under occlusion for 2 wk (three times per week). Topical 17β-estradiol was found to increase the expression of type 1 procollagen mRNA and protein significantly in human aged skin in vivo. In addition, MMP-1 protein levels were reduced by topical 17β-estradiol. The expressions of TGF-β1,TGF-βtype II receptor, and Sma and Mad related (Smad)3 were increased by topical 17β-estradiol in aged human skin, and TGF-β1 neutralizing antibody inhibited 17β-estradiol-induced procollagen synthesis in cultured fibroblasts. We also found that the expressions of tropoelastin and fibrillin-1 mRNA and protein, and elastic fibers in aged skin were also increased by topical 17β-estradiol. Topical 17β-estradiol also increased keratinocyte proliferation and the epidermal thickness in aged human skin. We also observed the same effects of topical 17β-estradiol in young skin. In conclusion, our results suggest that topical 17β-estradiol treatment may improve the cutaneous function of aged human skin by improving the connective tissue and increasing epidermal thickness.
