藁本内酯衍生物对软骨细胞外基质合成与分解代谢影响  

Effects of ligusticum cycloprolactam on synthesis and catabolism of extracellular matrix in chondrocytes

作　　者：刘建军 陈欣 汪欣月[3] 齐伟 王多贤 席芳琴 李宁 柳军玺 谢兴文[1,2] LIU Jianjun;CHEN Xin;WANG Xinyue;QI Wei;WANG Duoxian;XI Fangqin;LI Ning;LIU Junxi;XIE Xingwen(Affiliated Hospital of Gansu University of Traditional Chinese Medicine,Lanzhou 730000,China;Gansu University of Traditional Chinese Medicine,Lanzhou 730000,China;Beijing University of Chinese Medicine,Beijing 100029,China;Lanzhou Institute of Chemical Physics,Chinese Academy of Sciences,Lanzhou 730000,China)

机构地区：[1]甘肃中医药大学附属医院,甘肃兰州730000 [2]甘肃中医药大学,甘肃兰州730000 [3]北京中医药大学,北京100029 [4]中国科学院兰州化学物理研究所,甘肃兰州730000

出　　处：《中国骨质疏松杂志》2023年第11期1615-1620,共6页Chinese Journal of Osteoporosis

基　　金：甘肃省青年科技基金计划(20JR10RA345);兰州市城关区科技计划项目(2020-2-2-7);甘肃中医药大学附属医院院内创新基金(gzfy-2019-12);甘肃省科技计划资助(20JR5RA129);兰州市科技发展指导性计划项目(2020-ZD-65),甘肃省拔尖领军人才项目。

摘　　要：目的探究藁本内酯衍生物(LIGc)对IL-1β诱导大鼠软骨细胞外基质合成与分解代谢的影响。方法将大鼠软骨细胞分为空白对照组、IL-1β组及LIGc高剂量组(0.4μmol/mL)、中剂量组(0.2μmol/mL)、低剂量组(0.1μmol/mL)组。CCK-8法检测细胞活力;细胞免疫荧光检测COL2α1和ACAN的表达;采用RT-qPCR检测软骨细胞中SOX9、PRG4、MMP-13、ADAMTS-5的基因表达水平。Western blot检测软骨细胞中HMGB1、COX2、TLR4、NF-κB p65的蛋白表达水平。结果与正常组比较,IL-1β组软骨细胞增殖降低;COL2α1和ACAN的表达降低;SOX9、PRG4 mRNA表达下调,同时MMP-13、ADAMTS-5 mRNA表达上调;HMGB1、COX2、TLR4、NF-κB p65蛋白表达升高。与IL-1β组比较,LIGc能够促进软骨细胞的增殖;增强COL2α1和ACAN的表达;上调SOX9、PRG4 mRNA表达,同时下调MMP-13、ADAMTS-5 mRNA表达;下调HMGB1、COX2、TLR4、NF-κB p65蛋白表达。结论LIGc能够抑制IL-1β所致炎症因子表达,上调软骨细胞外基质合成代谢因子、下调分解代谢因子延缓软骨细胞外基质降解的作用,其机制可能与OA免疫炎症反应的TLR4/NF-κB信号通路相关。Objective To investigate the effect of ligusticum cycloprolactam(LIGc)on IL-1β-induced extracellular matrix synthesis and catabolism in rat cartilage.Methods Rat chondrocytes were divided into blank control group,IL-1βgroup and LIGc high-dose(0.4μmol/mL),medium-dose(0.2μmol/mL)and low-dose(0.1μmol/mL)groups;cell viability was detected by CCK-8 assay;cell immunofluorescence was performed to detect the expression of COL2α1 and ACAN;and chondrocyte gene expression levels were detected by RT-qPCR.gene expression levels of SOX9,PRG4,MMP-13 and ADAMTS-5 in the cells.The protein expression levels of HMGB1,COX2,TLR4,and NF-κB p65 in chondrocytes were detected by Western blot.Results Compared with the normal group,chondrocyte proliferation was reduced in the IL-1βgroup;the expression of COL2α1 and ACAN was decreased;the mRNA expression of SOX9 and PRG4 was downregulated,whereas the mRNA expression of MMP-13 and ADAMTS-5 was upregulated;and the protein expression of HMGB1,COX2,TLR4 and NF-κB p65 was increased.Compared with the IL-1βgroup,LIGc was able to promote the proliferation of chondrocytes;increase the expression of COL2α1 and ACAN.The expression of SOX9 and PRG4 mRNA was upregulated,while the expression of MMP-13 and ADAMTS-5 mRNA was downregulated.The protein expression of HMGB1,COX2,TLR4 and NF-κB p65 was downregulated.Conclusion LIGc can inhibit the expression of inflammatory factors induced by IL-1β,upregulate the anabolic factor of chondrocyte extracellular matrix and down-regulate the catabolic factor to delay the degradation of chondrocyte extracellular matrix.The mechanism may be related to the TLR4/NF-κB signalling pathway of the immune inflammatory response in osteoarthritis.

关 键 词：骨关节炎 藁本内酯衍生物 软骨细胞外基质 白细胞介素-1Β 

分 类 号：R336[医药卫生—人体生理学]