重组人结缔组织生长因子对人成骨细胞增殖、分化及凋亡的影响

目的 研究重组人结缔组织生长因子(rCTGF)对人成骨细胞增殖、分化及凋亡的影响.方法 用rCTGF干预体外培养的人成骨细胞,采用[3H]-TdR掺入法检测人成骨细胞增殖率,α-磷酸萘酚法测定细胞内碱性磷酸酶活性变化,放射免疫法测定骨钙素含量的变化,Western印迹法检测Ⅰ型胶原表达的变化,应用流式细胞仪检测rCTGF埘成骨细胞凋亡的影响.结果 rCTGF呈剂量依赖性促进人成骨细胞增殖,200 ng/ml rCTGF达最大效应;rCrrGF干预显著促进人成骨细胞分化,rCTGF旱剂量依赖性增加Ⅰ型胶原、骨钙素表达及碱件磷酸酶活性;rCTGF干预可显著减少成骨细胞凋亡.结论 rCTGF可显著促进人成骨细胞增殖、分化,并抑制人成骨细胞凋亡.

Effects of recombinant human connective tissue growth factor on proliferation, differentiation, and apoptosis in human osteoblasts

Objective To investigate the effects of recombinant human connective tissue growth factor (rCTGF) on the proliferation, differentiation and apoptosis in human osteoblasts. Methods Human osteoblasts were treated with rCTGF, cell proliferation was measured by [3 H] -thymidine ([3 H] -TdR) incorporation method,the activity of alkaline phosphatase (ALP) was determined using α-nitrophenyl phosphate as a substrate, the expression of type Ⅰ collagen was determined by Western blotting, osteocalcin in conditioned media was measured by radioimmune assay, and cell apoptosis was assayed by flow cytometry. Results rCTGF promoted proliferation and differentiation of human osteoblasts in a dose-depentent manner, rCTGF increased ALP activity, osteocalcin and type I collagen secretion in a dose-depentent manner, rCTGF inhibited human osteoblastic apoptosis induced by serum deprivation. Conclusion rCTGF can promote osteoblastic proliferation, and differentiation and protect osteoblast against apoptosis, suggesting that CTGF plays an essential role in bone metabolism.