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目的探讨补肾强脊颗粒对强直性脊柱炎(AS)的作用机制。方法以体外培养的AS髋关节囊组织的成纤维细胞为实验对象,分正常组,AS对照组,AS诱导组,兔空白血清组,补肾强脊颗粒小、中、大剂量组和SASP对照组。在细胞因子骨形态发生蛋白(BMP)与转化生长因子-β(TGF-β)的诱导刺激下,观察补肾强脊颗粒对细胞因子诱导后的AS成纤维细胞增殖(样本5个,采用酸性磷酸酶法)、细胞周期(样本3个,采用PI法流式细胞术)及细胞凋亡率(样本3个,采用Annexin V法流式细胞术)的影响。结果补肾强脊颗粒含药血清作用后,对细胞因子诱导下的AS成纤维细胞增殖有明显的抑制作用(P<0.01);使细胞因子诱导下的AS成纤维细胞G0/G1期细胞比例显著增加(P<0.05),S期的细胞比例降低(P<0.01);促使细胞因子诱导下的AS成纤维细胞早期凋亡率、晚期凋亡率均有不同程度的增加(P<0.01)。结论补肾强脊颗粒可以通过抑制细胞因子对成纤维细胞增殖、细胞周期及凋亡的影响而达到抑制成纤维细胞进一步向成骨型的分化。
Objective To investigate the mechanism of Bushen Jiangji granule on ankylosing spondylitis (AS). Methods Fibroblasts from AS hip joint capsule cultured in vitro were divided into three groups: normal control group, AS control group, AS induction group, blank rabbit serum group, Bushenjiangji Granule small and medium dose group and SASP control group . Under the stimulation of bone morphogenetic protein (BMP) and transforming growth factor-β (TGF-β), we observed the proliferation of AS-induced fibroblasts induced by cytokine (5 samples, Enzyme method), cell cycle (3 samples, using PI method flow cytometry) and apoptosis rate (3 samples, using Annexin V method flow cytometry). Results Bushenqiangji granule could significantly inhibit the proliferation of AS fibroblasts induced by cytokines (P <0.01), and the proportion of cells in G0 / G1 phase induced by cytokines was significant (P <0.05), and the proportion of cells in S phase decreased (P <0.01). The apoptosis rate of AS fibroblasts induced by cytokines increased to some extent (P <0.01). Conclusion Bushen Jiangji granules can inhibit fibroblasts from further differentiating into osteoblasts by inhibiting the effects of cytokines on the proliferation, cell cycle and apoptosis of fibroblasts.