CTGF对破骨前体细胞RAW264.7增殖及碳酸酐酶Ⅱ分化的影响

Effect of CTGF on carbonic anhydrase II and the proliferation and the differentiation of RAW264.7 cells

目的研究结缔组织生长因子(CTGF)对体外培养的破骨细胞前体细胞RAW264.7增殖及对核因子Kappa B配体受体(RANKL)诱导体外培养的破骨细胞前体细胞RAW264.7分化为成熟多核破骨细胞的影响。方法使用200 ng/mLCTGF干预培养的破骨细胞前体细胞RAW264.7,采用3H-TdR掺入法检测RAW264.7细胞增殖率;使用200 ng/mL CTGF与RANKL单独或共同处理RAW264.7细胞,抗酒石酸酸性磷酸酶(TRAP)染色观察TRAP阳性多核细胞,Western blot检测碳酐酶Ⅱ蛋白的表达。结果 CTGF可显著促进RAW264.7细胞增殖;200 ng/mLCTGF与RANKL共同处理RAW264.7细胞可促进RAW264.7细胞分化为成熟多核破骨细胞;200 ng/mL CTGF与RANKL共同处理RAW264.7细胞可促进RAW264.7细胞碳酐酶Ⅱ蛋白的表达。结论 CTGF促进体外培养的破骨细胞前体细胞RAW264.7增殖,促进RANKL诱导的破骨细胞前体细胞RAW264.7分化为成熟多核破骨细胞。

Objective To investigate the effect of CTGF on the proliferation of cultured RAW264.7 cells and the differentiation of mature multinuclear osteoclasts induced by RANKL. Methods RAW264. 7 cells were cultured with the intervention of 200ng/ ml CTGF. The cell proliferation rate was detected using （^3H）-thymidine （^3H-TdR） incorporation method. After the intervention with or without RANKL, TRAP staining was performed to observe TRAP-positive multinuclear cells （MNCs）. The expression of carbonic anhydrase II was detected using Western blotting. Results CTGF could significantly promote the proliferation of RAW264.7 cells. The intervention of 200ng/ml CTGF combined with RANKL could promote RAW264. 7 cell differentiation into mature multinuclear osteoclasts. It also could promote the expression of carbonic anhydrase II in RAW264. 7 cells. Conclusion CTGF canpromote the proliferation of cultured RAW264.7 cells and promote its differentiation into mature multinuclear osteoclasts induced by RANKL.