Objective： To evaluate the effect of Zhuanggu Jianxi Decoction （壮骨健膝方, ZGJXD） on interleukin- 1 β- （IL-1 β- ）-induced degeneration of chondrocytes （CDs） as well as the activation of caveolin-p38 mitogen-...Objective： To evaluate the effect of Zhuanggu Jianxi Decoction （壮骨健膝方, ZGJXD） on interleukin- 1 β- （IL-1 β- ）-induced degeneration of chondrocytes （CDs） as well as the activation of caveolin-p38 mitogen- activated protein kinase （MAPK） signal pathway, investigating the possible molecular mechanism that ZGJXD treats osteoarthritis. Methods： Serum pharmacology was applied in the present study, where ZGJXD was orally administrated to New Zealand rabbits and then ZGJXD containing serum （ZGJXD-S） was collected for following in vitro experiments. CDs were isolated aseptically from New Zealand rabbits and then cultured in vitro. Upon IL- l β-stimulation, the degeneration of CDs was verified by inverted microscope, toluidine blue stain and type H collagen immunocytochemistry. After IL-1 β-stimulated CDs were intervened with blank control serum, ZGJXD-S, together with or without SB203580 （a specific inhibitor of p38 MAPK） for 48 h, caveoUn-1 protein expression and the phosphorylation level of p38 were determined by Western blotting, and the mRNA expression of IL- 1β-, tumor necrosis factor ~ （TNF-oL）, matrix metalloproteinase 3 （MMP-3） and MMP-β were examined by real-time polymerase chain reaction. Results： IL-1 β- stimulation induced degeneration of CDs, increased caveolin-1 expression and p38 phosphorylation, up-regulated the mRNA level of IL-1 β, TNF-β, MMP-3 and MMP-β. However, the IL-1 β-induced activation of caveolin-p38 signaling and alteration in the expression of p38 downstream target genes were suppressed by ZGJXD-S and/or SB203580 in CDs. Conclusion： ZGJXD can prevent CDs degeneration via inhibition of caveolin-p38 MAPK signal pathway, which might be one of the mechanisms that ZGJXD treats osteoarthritis.展开更多
背景:软骨细胞退变常导致骨关节炎、椎间盘退变、半月板退变等骨科常见疾病,沉默信息调节因子1(silent information regulator 1,SIRT1)基因与软骨细胞退变关系密切,当敲减ATDC5小鼠软骨细胞系中SIRT1基因后,可通过软骨细胞退变相关信...背景:软骨细胞退变常导致骨关节炎、椎间盘退变、半月板退变等骨科常见疾病,沉默信息调节因子1(silent information regulator 1,SIRT1)基因与软骨细胞退变关系密切,当敲减ATDC5小鼠软骨细胞系中SIRT1基因后,可通过软骨细胞退变相关信号通路的作用影响细胞功能,进而影响骨科疾病的进程。目的:探讨SIRT1基因敲减后ATDC5小鼠软骨细胞中显著激活的软骨细胞退变相关信号通路情况及差异因子的表达情况。方法:携载或不携载SIRT1基因的慢病毒转染ATDC5细胞后,分为对照组(转染携载阴性对照慢病毒ATDC5细胞组)和实验组(转染携载SIRT1基因慢病毒ATDC5细胞组)。提取不同组别细胞中总RNA并进行质检,基因芯片检测细胞中编码RNA核酸的表达情况,结合生物信息学分析,寻找显著激活的软骨细胞退变相关信号通路,并对其中部分信号通路的差异变化因子进行阐述。结果与结论:①发现SIRT1基因敲减后ATDC5细胞中被显著激活的信号通路有42条;②选择了其中被显著激活的5条与软骨细胞退变相关且研究较少的信号通路:肝细胞生长因子信号通路(ETS1、PIK3CA、NRAS、PIK3C2A、FGFR2、ELF1、FGFR3、CDKN1A、AKT3、FRS2、PRKD3、MAP3K2)(Ratio=0.1390,Z-score=2.673)、神经调节因子信号通路(RPS6KB1、STAT5A、MTOR、BTC、HBEGF、RNF41)(Ratio=0.1360,Z-score=2.309)、胰岛素受体信号通路(TSC1、EIF4EBP1、PRKAR2B、PPP1R12A、TSC2)(Ratio=0.1060,Z-score=2.138)、趋化因子受体4信号通路(PAK4、EGR1、RHOJ、RHOB、PLCB1、GNG5)(Ratio=0.0970,Z-score=2.500)、肌动蛋白细胞骨架信号通路(ABI2、BRK1、PIP4K2B、MYLK、IQGAP1、ARPC1A、ACTA2、EZR、SSH2、ACTG1、IQGAP3)(Ratio=0.0921,Z-score=2.236)进行汇报,从而为研究软骨细胞退变相关疾病提供新颖靶点。展开更多
基金Supported by the National Natural Science Foundation of China(No.81072828)
文摘Objective： To evaluate the effect of Zhuanggu Jianxi Decoction （壮骨健膝方, ZGJXD） on interleukin- 1 β- （IL-1 β- ）-induced degeneration of chondrocytes （CDs） as well as the activation of caveolin-p38 mitogen- activated protein kinase （MAPK） signal pathway, investigating the possible molecular mechanism that ZGJXD treats osteoarthritis. Methods： Serum pharmacology was applied in the present study, where ZGJXD was orally administrated to New Zealand rabbits and then ZGJXD containing serum （ZGJXD-S） was collected for following in vitro experiments. CDs were isolated aseptically from New Zealand rabbits and then cultured in vitro. Upon IL- l β-stimulation, the degeneration of CDs was verified by inverted microscope, toluidine blue stain and type H collagen immunocytochemistry. After IL-1 β-stimulated CDs were intervened with blank control serum, ZGJXD-S, together with or without SB203580 （a specific inhibitor of p38 MAPK） for 48 h, caveoUn-1 protein expression and the phosphorylation level of p38 were determined by Western blotting, and the mRNA expression of IL- 1β-, tumor necrosis factor ~ （TNF-oL）, matrix metalloproteinase 3 （MMP-3） and MMP-β were examined by real-time polymerase chain reaction. Results： IL-1 β- stimulation induced degeneration of CDs, increased caveolin-1 expression and p38 phosphorylation, up-regulated the mRNA level of IL-1 β, TNF-β, MMP-3 and MMP-β. However, the IL-1 β-induced activation of caveolin-p38 signaling and alteration in the expression of p38 downstream target genes were suppressed by ZGJXD-S and/or SB203580 in CDs. Conclusion： ZGJXD can prevent CDs degeneration via inhibition of caveolin-p38 MAPK signal pathway, which might be one of the mechanisms that ZGJXD treats osteoarthritis.
文摘背景:软骨细胞退变常导致骨关节炎、椎间盘退变、半月板退变等骨科常见疾病,沉默信息调节因子1(silent information regulator 1,SIRT1)基因与软骨细胞退变关系密切,当敲减ATDC5小鼠软骨细胞系中SIRT1基因后,可通过软骨细胞退变相关信号通路的作用影响细胞功能,进而影响骨科疾病的进程。目的:探讨SIRT1基因敲减后ATDC5小鼠软骨细胞中显著激活的软骨细胞退变相关信号通路情况及差异因子的表达情况。方法:携载或不携载SIRT1基因的慢病毒转染ATDC5细胞后,分为对照组(转染携载阴性对照慢病毒ATDC5细胞组)和实验组(转染携载SIRT1基因慢病毒ATDC5细胞组)。提取不同组别细胞中总RNA并进行质检,基因芯片检测细胞中编码RNA核酸的表达情况,结合生物信息学分析,寻找显著激活的软骨细胞退变相关信号通路,并对其中部分信号通路的差异变化因子进行阐述。结果与结论:①发现SIRT1基因敲减后ATDC5细胞中被显著激活的信号通路有42条;②选择了其中被显著激活的5条与软骨细胞退变相关且研究较少的信号通路:肝细胞生长因子信号通路(ETS1、PIK3CA、NRAS、PIK3C2A、FGFR2、ELF1、FGFR3、CDKN1A、AKT3、FRS2、PRKD3、MAP3K2)(Ratio=0.1390,Z-score=2.673)、神经调节因子信号通路(RPS6KB1、STAT5A、MTOR、BTC、HBEGF、RNF41)(Ratio=0.1360,Z-score=2.309)、胰岛素受体信号通路(TSC1、EIF4EBP1、PRKAR2B、PPP1R12A、TSC2)(Ratio=0.1060,Z-score=2.138)、趋化因子受体4信号通路(PAK4、EGR1、RHOJ、RHOB、PLCB1、GNG5)(Ratio=0.0970,Z-score=2.500)、肌动蛋白细胞骨架信号通路(ABI2、BRK1、PIP4K2B、MYLK、IQGAP1、ARPC1A、ACTA2、EZR、SSH2、ACTG1、IQGAP3)(Ratio=0.0921,Z-score=2.236)进行汇报,从而为研究软骨细胞退变相关疾病提供新颖靶点。
