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Bushen-Qiangdu-Zhilv decoction inhibits osteogenic differentiation of rat fibroblasts by regulating connexin 43

机译:补肾强毒汤通过调节连接蛋白43抑制大鼠成纤维细胞的成骨分化

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摘要

Bushen-Qiangdu-Zhilv (BQZ) decoction is a traditional Chinese medicinal compound widely used for treating ankylosing spondylitis (AS). However, the mechanisms underlying effects of BQZ remain largely unknown. Osteoblast differentiation of fibroblasts plays an important role in heterotopic ossification (HO) of AS, and connexin 43 (Cx43) is crucially involved in the osteoblast differentiation of fibroblasts. The aim of the present study was to evaluate the effects of BQZ on the osteogenic differentiation of fibroblasts by regulating Cx43. Rat fibroblasts were treated with freeze-dried powder of BQZ, in the presence or absence of recombinant human bone morphogenetic protein-2 (rhBMP-2). MTS assays were performed to examine the inhibitory effects of BQZ on fibroblast proliferation. Western blot assays were conducted to detect the protein expression of core-binding factor alpha 1 (Cbfα1), Cx43 and phosphorylated Cx43 (pCx43). BQZ appeared to inhibit fibroblast proliferation in a dose-dependent manner. Furthermore, the expression of Cbfα1 and Cx43/pCx43 was significantly suppressed by BQZ, with or without rhBMP-2 stimulation. Therefore, the present results indicate that BQZ may exert an anti-AS effect by suppressing the osteogenic differentiation of fibroblasts via Cx43 regulation.
机译:补肾强身汤(BQZ)是一种中药,广泛用于治疗强直性脊柱炎(AS)。但是,BQZ的潜在作用机理仍然未知。成纤维细胞的成骨细胞分化在AS的异位骨化(HO)中起着重要作用,而连接蛋白43(Cx43)至关重要地参与了成纤维细胞的成骨细胞分化。本研究的目的是通过调节Cx43评估BQZ对成纤维细胞成骨分化的影响。在有或没有重组人骨形态发生蛋白2(rhBMP-2)的情况下,将BQZ冻干粉处理大鼠成纤维细胞。进行MTS测定以检查BQZ对成纤维细胞增殖的抑制作用。进行蛋白质印迹分析以检测核心结合因子α1(Cbfα1),Cx43和磷酸化的Cx43(pCx43)的蛋白表达。 BQZ似乎以剂量依赖的方式抑制成纤维细胞的增殖。此外,在有或没有rhBMP-2刺激的情况下,BQZ均可显着抑制Cbfα1和Cx43 / pCx43的表达。因此,本结果表明,BQZ可通过经由Cx43调节来抑制成纤维细胞的成骨分化而发挥抗AS作用。

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