摘要
目的:研究枳壳甘草汤对大鼠椎间盘退变(IDD)及髓核组织生物钟相关基因表达的影响。方法:60只SD大鼠随机分成假手术组、模型组、塞来昔布组及枳壳甘草汤低、中、高剂量组,每组10只。大鼠尾椎间盘针刺构建IDD模型。各组予相应药物干预后取材,X线扫描及病理染色检测大鼠尾椎间盘病理改变,酶联免疫吸附法检测尾椎间盘组织炎症因子白细胞介素(IL)-1β、IL-4、IL-6、肿瘤坏死因子-α(TNF-α)水平,实时荧光定量PCR检测椎间盘髓核组织生物钟相关mRNA表达水平,Western Blot检测椎间盘髓核组织细胞外基质及生物钟相关蛋白表达水平。结果:与假手术组比较,模型组尾椎间盘高度指数显著降低(P<0.01),病理改变明显,椎间盘IL-1β、IL-6、TNF-α及ADAMTS4、MMP3蛋白表达显著升高(P<0.01),IL-4,Aggrecan、CollagenⅡ蛋白及Bmal1、Clock、Rev-erbα、Rorα、Per1、Cry1 mRNA和蛋白表达显著降低(P<0.01)。与模型组比较,塞来昔布组及枳壳甘草汤中、高剂量组大鼠尾椎间盘高度指数显著升高(P<0.01),病理改变减轻,椎间盘IL-1β、IL-6、TNF-α及ADAMTS4、MMP3蛋白表达显著降低(P<0.01),IL-4及Aggrecan、CollagenⅡ蛋白表达显著升高(P<0.01,P<0.05),塞来昔布组Clock、Rev-erbα、Rorα、Per1、Cry1 mRNA及蛋白表达显著升高(P<0.01,P<0.05),枳壳甘草汤高剂量组Bmal1、Clock、Rev-erbα、Rorα、Per1、Cry1 mRNA及蛋白表达显著升高(P<0.01)。结论:枳壳甘草汤可以抑制退变椎间盘髓核组织炎症及细胞外基质降解,延缓IDD,内在机制可能与其对生物钟基因的调控有关。
Objective:To investigate the effects of Zhiqiao Gancao Decoction on intervertebral disc degeneration(IDD)and the expression of circadian clock-related genes in the nucleus pulnosus tissue of a rat model.Methods:Sixty SD rats were randomly divided into sham group,model group,celecoxib group and Zhiqiao Gancao Decoction low-dose,medium-dose and high-dose groups,with 10 rats in each group.IDD model was constructed by needle puncture of the rat tail intervertebral disc.After respective interventions,samples were taken from each group.X-ray scanning and pathological staining were performed to detect pathological changes in the rat tail intervertebral disc.Enzyme-linked immunosorbent assay was used to detect the levels of inflammatory factors interleukin(IL)-1β,IL-4,IL-6,tumor necrosis factor-alpha(TNF-α)in the intervertebral disc tissue.Quantitative Real-time PCR was used to detect the expression levels of clock-related genes mRNA in the nucleus pulposus tissue,and Western Blot was used to detect the expression levels of extracellular matrix and circadian clock-related proteins in the nucleus pulposus tissue.Results:Compared with the sham group,the model group showed a significant decrease in intervertebral disc height index(P<0.01),significant pathological changes in the intervertebral disc,significant increase in the expression of IL-1β,IL-6,TNF-α,ADAMTS4,MMP3 proteins(P<0.01),significant decrease in the expression of IL-4,Aggrecan,Collagen II proteins,and Bmall,Clock,Rev-erba,Rora,Perl,Cryl mRNA and proteins(P<0.01).Compared with the model group,the celecoxib group and the Zhiqiao Gancao Decoction medium-dose and high-dose groups showed a significant increase in intervertebral disc height index(P<0.01),alleviation of intervertebral disc pathological changes,significant decrease in the expression of IL-1β,IL-6,TNF-α,ADAMTS4,MMP3 proteins(P<0.01),significant increase in the expression of IL-4,Aggrecan,Collagen I proteins(P<0.01,P<0.05).In the celecoxib group,the expresion of Clock,Rev-erba,Rora,Perl,Cryl mRNA and proteins were significantly increased(P<0.01,P<0.05);in the Zhiqiao Gancao Decoction high-dose group,the expression of Bmall,Clock,Rev-erba,Rora,Perl,Cryl mRNA and proteins were significantly increased(P<0.01).Conclusion:Zhiqiao Gancao Decoction can inhibit the inflammation of nucleus pulposus tissueand the degradation of extracelular matrix,and delay IDD,the internal mechanism may be related to the regulation of circadian clock genes.
作者
黄泽灵
徐波
陆斌杰
蔡学峰
裴帅
刘昱江
祝学思
沈晓峰
李宇卫
HUANG Zeling;XU Bo;LU Binjie;CAI Xuefeng;PEI Shuai;LIU Yujiang;ZHU Xuesi;SHEN Xiaofeng;LI Yuwei(Suzhou TCM Hospital Affiliated to Nanjing University of Chinese Medicine,Suzhou 215009,China;Orthopaedic Traumatology Institute,Suzhou Academy of Wumen Chinese Medicine,Suzhou 215009,China)
出处
《中华中医药杂志》
北大核心
2025年第2期516-523,共8页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金面上项目(No.82174399)
苏州市科技发展计划项目(No.SKYD2023150)
江苏省研究生培养创新工程研究生实践创新计划(No.SJCX24_1023)
江苏省中医药管理局重点项目(No.ZD202230)
苏州市中医药管理局临床重点项目(No.LCZX202320)
中医骨伤科学重点实验室建设项目(No.JSDW202253,No.SZS2022019)
江苏省医学重点学科/实验室建设单位(No.苏卫科教[2022]17号)。
关键词
腰痛
椎间盘退变
生物钟
炎症因子
细胞外基质
枳壳甘草汤
Low back pain
Intervertebral disc degeneration(IDD)
Circadian clock
Inflammatory factors
Extracellular matrix
ZhiqiaoGancaoDecoction
