补肾活血方抑制骨关节炎大鼠软骨细胞凋亡的机制  

The mechanism of Bushen Huoxue formula in the inhibition of chondrocyte apoptosis in rats with osteoarthritis

作　　者：胡宏志[1] 李娟[1] 毛滔[1] HU Hongzhi;LI Juan;MAO Tao(Department of Orthopedics and Trauma,the Second Affiliated Hospital of Hunan University of Traditional Chinese Medicine,Changsha 410005,China)

机构地区：[1]湖南中医药大学第二附属医院骨伤四科,湖南长沙410005

出　　处：《中国骨质疏松杂志》2025年第3期382-387,396,共7页Chinese Journal of Osteoporosis

基　　金：湖南省中医药科研计划项目(D2022114);湖南省自然科学基金省市联合项目(2022JJ50049)。

摘　　要：目的探讨补肾活血方(BSHX)抑制骨关节炎大鼠软骨细胞凋亡的作用机制。方法将CHON-001细胞随机分为空白组、LPS组(100 ng/mL)、BSHX组(100 ng/mL LPS+20%BSHX含药血清)、BSHX+DMSO组(100 ng/mL LPS+20%BSHX含药血清+0.1%DMSO)、BSHX+Derivative 83组(100 ng/mL LPS+20%BSHX含药血清+120μmol/L Derivative 83)。分析各组细胞增殖、凋亡以及上清液中肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6、IL-1β水平;大鼠随机分为假手术组、模型组、BSHX组,检测血清中一氧化氮合成酶(INOS)、IL-6、环氧合酶2(COX2)水平以及软骨组织中CollageⅡ、Aggrecan、ADAMTS-5 mRNA表达水平;并对软骨组织及细胞中Cleaved caspase-3、Bax、Bcl-2及Wnt3a/β-catenin蛋白表达水平进行分析。结果与空白组相比,LPS组细胞凋亡率、TNF-α、IL-6、IL-1β水平、Cleaved caspase-3、Bax、Wnt3a、β-catenin、p-NF-kB p65/NF-kB p65蛋白表达显著增加,24 h、48 h OD450值以及EdU阳性率、Bcl-2表达明显降低(P<0.05);BSHX改善了LPS诱导的CHON-001细胞上述指标(P<0.05),但Derivative 83逆转了BSHX对LPS诱导的CHON-001细胞的保护作用(P<0.05)。动物实验结果显示,与假手术组相比,模型组INOS、IL-6、COX2水平、ADAMTS-5 mRNA、Cleaved caspase-3、Bax、Wnt3a、β-catenin、p-NF-kB p65/NF-kB p65蛋白表达显著增加,CollageⅡmRNA、Aggrecan mRNA、Bcl-2表达明显降低(P<0.05);与模型组相比,BSHX组INOS、IL-6、COX2水平、ADAMTS-5 mRNA、Cleaved caspase-3、Bax、Wnt3a、β-catenin、p-NF-kB p65/NF-kB p65蛋白表达显著降低,CollageⅡmRNA、Aggrecan mRNA、Bcl-2表达明显增加(P<0.05)。结论BSHX抑制骨关节炎大鼠软骨细胞凋亡。Objective To investigate the mechanism of Bushen Huoxue formula(BSHX)in inhibiting apoptosis of chondrocytes in rats with osteoarthritis.Methods CHON-001 cells were randomly grouped into blank group,LPS group(100 ng/mL),BSHX group(100 ng/mL LPS+20%BSHX medicated serum),BSHX+DMSO group(100 ng/mL LPS+20%BSHX medicated serum+0.1%DMSO),and BSHX+Derivative 83 group(100 ng/mL LPS+20%BSHX medicated serum+120μmol/L Derivative 83).The proliferation and apoptosis of cells,and the levels of tumor necrosis factor(TNF)-α,interleukin(IL)-6 and IL-1βin the supernatant in each group were analyzed.The rats were randomly divided into sham operation group,model group,and BSHX group.The serum levels of nitric oxide synthase(INOS),IL-6,and cyclooxygenase 2(COX2),and the mRNA expression levels of Collage II,Aggrecan and ADAMTS-5 in cartilage tissue were measured.The expression levels of Cleaved caspase-3,Bax,Bcl-2,and Wnt3a/β-catenin in the cartilage and cells were analyzed.Results Compared to those in the blank group,the apoptosis rate,TNF-α,IL-6,IL-1βlevels,Cleaved caspase-3,Bax,Wnt3a,β-catenin,and p-NF-kB p65/NF-kB p65 protein expressions in the LPS group increased obviously.The OD 450 value at 24 h and 48 h,the positive rate of EdU,and the expression of Bcl-2 decreased obviously(P<0.05).BSHX improved the above indexes in LPS-induced CHON-001 cells(P<0.05).Derivative 83 reversed the protective effect of BSHX on LPS-induced CHON-001 cells(P<0.05).The result of animal experiments showed that compared to those in the sham group,the protein expressions of INOS,IL-6,and COX2,ADAMTS-5 mRNA,Cleaved caspase-3,Bax,Wnt3a,β-catenin,and p-NF-kB p65/NF-kB p65 in the model group increased obviously,and the expressions of collage II mRNA,aggrecan mRNA,and Bcl-2 decreased obviously(P<0.05).Compared to those in the model group,the protein expressions of INOS,IL-6,COX2,ADAMTS-5 mRNA,Cleaved caspase-3,Bax,Wnt3a,β-catenin,and p-NF-kB p65/NF-kB p65 decreased obviously in the BSHX group,the expressions of collage II mRNA,aggrecan mRNA and Bcl-2

关 键 词：补肾活血方 骨关节炎 软骨细胞 凋亡 

分 类 号：R285.5[医药卫生—中药学]