温肾通络止痛方对炎性骨丢失小鼠MAPK信号通路的影响  

Effects of Wenshen Tongluo Zhitong Decoction on inflammatory bone loss in mice by regulating MAPK signaling pathway

作　　者：韩秋革 余自层 郭杨[1] 潘娅岚 刘孟敏 孙杰[1] 王礼宁 员浬 马勇[1,2] HAN Qiuge;YU Ziceng;GUO Yang;PAN Yalan;LIU Mengmin;SUN Jie;WANG Lining;YUN Li;MA Yong(Laboratory of Traumatology&Orthopedics,Nanjing University of Chinese Medicine,Nanjing 210023,China;School of Chinese Medicine·School of Integrated Chinese and Western Medicine,Nanjing University of Chinese Medicine,Nanjing 210023,China;School of Nursing,Nanjing University of Chinese Medicine,Nanjing 210023,China)

机构地区：[1]南京中医药大学骨伤修复与重建新技术实验室,南京210023 [2]南京中医药大学中医学院·中西医结合学院,南京210023 [3]南京中医药大学护理学院,南京210023

出　　处：《中华中医药杂志》2023年第10期4694-4699,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金面上项目(No.82074458,No.82174411);江苏省自然科学基金面上项目(No.BK20221351);江苏高校中西医临床医学品牌专业建设工程资助项目(No.2020PPZXL261);江苏省科研与实践创新计划项目(No.KYCX22_1882)。

摘　　要：目的:观察温肾通络止痛方对炎性骨丢失小鼠骨微结构和骨吸收的影响,探讨其抗炎性骨丢失的作用机制。方法:将50只小鼠随机分为正常组,模型组,阿仑膦酸钠组,温肾通络止痛方低、高剂量组,每组10只。腹腔注射脂多糖(5 mg/kg,1次/周)构建小鼠炎性骨丢失模型。采用HE和Micro CT方法观察股骨结构,ELISA法检测血清中IL-6、IL-1β、ALP、TRAP的表达量,RT-PCR法检测胫骨组织中NFATC1、CTSK mRNA的表达,Western blot法检测胫骨组织中JNK、ERK、p38、NFATC1、CTSK蛋白表达。结果:与模型组比较,温肾通络止痛方高、低剂量组均可改善骨小梁密度,减小骨髓腔间隙,缓解小鼠骨质流失;增加3D BMD、BV/TV、Tb.Th,降低Tb.Sp;降低血清中IL-1β、IL-6、ALP、TRAP的表达(P<0.01);降低NFATC1、CTSK mRNA的表达水平(P<0.05),并抑制NFATC1、CTSK、p38、ERK、JNK蛋白表达(P<0.01)。结论:温肾通络止痛方能够显著改善炎性介导的小鼠骨质流失,其作用机制可能与MAPK信号通路有关。Objective:To observe the effects of Wenshen Tongluo Zhitong Decoction(WTZD)on bone microstructure and bone resorption in mice with inflammatory bone loss,and to preliminarily explore the mechanism of its anti-inflammatory bone loss.Methods:Fifty mice were randomly divided into normal group,model group,alendronate sodium group,WTZD low dose,high dose groups,with 10 mice in each group.Mice inflammatory bone loss model was established by intraperitoneal injection of LPS(5 mg/kg,once a week).HE and Micro CT were used to observe the femur structure,ELISA was used to detect the expression levels of IL-6,IL-1β,ALP and TRAP in serum,and RT-PCR was used to detect the mRNA expressions of NFATCI and CTSK in tibia tissue.The protein expressions of JNK,ERK,p38,NFATC1 and CTSK in tibia were detected by Western blot.Results:Compared with model group,both high and low dose groups of WTZD could improve bone trabecular density,reduce bone marrow space,and alleviate bone loss in mice;WTZD also could increased 3D BMD,BV/TV,Tb.Th,and decreased Tb.Sp;the expressions of IL-1β,IL-6,ALP and TRAP in serum were decreased(P<0.01).The mRNA expression levels of NFATC1 and CTSK were decreased(P<0.05),and the protein expressions of NFATC1,CTSK,p38,ERK and JNK were inhibited(P<0.01).Conclusion:WTZD can significantly improve inflammation-mediated bone loss in mice,and its mechanism may be related to MAPK signaling pathway.

关 键 词：温肾通络止痛方 炎性骨丢失 巨噬细胞极化 破骨细胞分化 MAPK信号通路 

分 类 号：R285.5[医药卫生—中药学]