葛根素抑制强直性脊柱炎患者成纤维细胞的增殖和成骨分化

Puerarin inhibits the proliferation and ossification of fibroblasts in ankylosing spondylitis

目的探讨葛根素对强直性脊柱炎(ankylosing spondylitis,AS)患者成纤维细胞增殖和成骨分化的影响及其机制。方法从AS患者和股骨颈骨折患者髋关节囊组织中分离培养成纤维细胞,采用不同浓度的葛根素(25、50、100、150、200 mg/L)处理细胞,CCK-8检测细胞增殖,Western blot检测IL-6、TNF-α、骨钙素(OCN)和骨桥蛋白(OPN)的蛋白表达,检测细胞的ALP活性,qRT-PCR检测ColⅠ基因的mRNA表达。结果在正常成纤维细胞中,150 mg/L和200 mg/L的葛根素能够明显抑制细胞增殖。在AS成纤维细胞中,100、150、200 mg/L的葛根素对细胞增殖有明显抑制作用,并且200 mg/L的葛根素对两种细胞增殖的抑制作用最强。葛根素降低正常成纤维细胞和AS患者成纤维细胞中致炎性细胞因子IL-6和TNF-α的蛋白表达,减弱细胞ALP活性,降低ColⅠ基因的mRNA表达,并抑制OCN和OPN的蛋白表达。结论葛根素可以抑制正常成纤维细胞和AS患者成纤维细胞的增殖和成骨性分化。

Objective To explore the effect and underlying mechanism of puerarin on cell proliferation and ossification of fibroblasts in ankylosing spondylitis(AS).Methods Fibroblasts were isolated from the hip capsule tissue of AS patients or femoral neck fractures.The cells were treated with puerarin at different concentrations(25,50,100,150,and 200 mg/L).Cell proliferation was measured by CCK-8 method.Protein expressions of IL-6,TNF-α,OCN and OPN were measured by Western blot.ALP activity was detected and the mRNA expression of Col I was evaluated by qRT-PCR.Results Puerarin of 150 and 200 mg/L significantly inhibited the proliferation of normal fibroblasts,while puerarin of 100,150 and 200 mg/L significantly inhibited the proliferation of AS fibroblasts.Puerarin significantly inhibited protein expressions of pro-inflammatory cytokines IL-6 and TNF-α,decreased ALP activity,inhibited mRNA expression ofColⅠ,and attenuated protein expressions of OCN and OPN.Conclusion Puerarin inhibits cell proliferation and fibroblast ossification of normal and AS fibroblasts.