CTGF对破骨前体细胞RAW264.7增殖及碳酸酐酶Ⅱ分化的影响

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。

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

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 can promote the proliferation of cultured RAW264.7 cells and promote its differentiation into mature multinuclear osteoclasts induced by RANKL.