温经通络汤对膝骨关节炎模型小鼠VEGF/VEGFR2/ERK1/2信号通路的影响  

Effect of Wenjing Tongluo Decoction on VEGF/VEGFR2/ERK1/2 Signaling Pathway in Mice with Knee Osteoarthritis

作　　者：许奇 钱佳佳[1,3] 许炜民 黄桂成 马勇[1,2] 郭杨[1,2] 韩龙[1,2] 王礼宁 XU Qi;QIAN Jia-jia;XU Wei-min;HUANG Gui-cheng;MA Yong;GUO Yang;HAN Long;WANG Li-ning(Laboratory of New Techniques of Restoration&Reconstruction of Orthopedics and Traumatology,Nanjing University of Chinese Medicine,Nanjing 210023,China;Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China;School of Acupuncture-Moxibustion and Tuina,School of Regimen and Rehabilitation,Nanjing University of Chinese Medicine,Nanjing 210023,China)Objective:To investigate the possible mechanism of Wenjing Tongluo)

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]南京中医药大学附属医院,南京210029 [3]南京中医药大学针灸推拿学院·养生康复学院,南京210023

出　　处：《中国实验方剂学杂志》2021年第13期28-34,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金面上项目(81774341);全国老中医药专家学术经验继承工作资助项目(国中医药人教发[2017]29号);江苏高校自然科学研究面上项目(20KJB360004)。

摘　　要：目的:探讨温经通络汤改善膝骨关节炎关节(KOA)软骨缺损,延缓关节退变的可能作用机制。方法:采用前交叉韧带切除术(ACLT法)建立KOA模型,实验分为假手术组、模型组、温经通络汤组(高、低剂量)和阳性药组。造模4周后中药组分别予以高、低剂量(80,20 g·kg^(-1))的温经通络汤灌胃;阳性药组予以硫酸氨基葡萄糖胶囊(0.29 g·kg^(-1));假手术组与模型组均予等体积生理盐水灌胃。连续干预4周后苏木素-伊红(HE)染色观察软骨组织形态学变化并进行Mankin’s评分;蛋白免疫印迹法(Western blot)和实时荧光定量聚合酶链式反应(Real-time PCR)检测血管内皮生长因子A(VEGFA),血管内皮生长因子受体2(VEGFR2),细胞外调节蛋白激酶1/2(ERK1/2)和血小板反应蛋白解整合素金属肽酶4(ADAMTS4)蛋白和mRNA的表达。结果:HE染色结果显示与假手术组比较,模型组小鼠Mankin’s评分显著升高(P<0.01);与模型组比较,药物组均能明显改善造模引起的关节软骨面缺损,Mankin’s评分均有显著降低(P<0.01),但各给药组间差异无有统计学意义。与假手术组比较,模型组VEGFA,VEGFR2,ERK1/2和ADAMTS4蛋白和mRNA的表达显著升高(P<0.01)。与模型组比较,各给药组可以显著抑制VEGFA和ERK1/2的蛋白表达(P<0.01),硫酸氨基葡萄糖胶囊组效果要优于温经通络汤低剂量组(P<0.05),但弱于温经通络汤高剂量组(P<0.01);硫酸氨基葡萄糖胶囊组可以抑制VEGFR2和ADAMTS4的蛋白表达,作用与温经通络汤低剂量组相当,作用均弱于温经通络汤高剂量组(P<0.05)。结论:温经通络汤可以改善KOA小鼠关节软骨损伤,其作用机制可能与VEGF/VEGFR2/ERK1/2信号通路有关。Objective:To investigate the possible mechanism of Wenjing Tongluo decoction(WTD)in alleviating articular cartilage defect in knee osteoarthritis(KOA)and delaying joint degeneration.Method:The KOA model was established by anterior cruciate ligament transection(ACLT).Mice were classified into shamoperated group,model group,WTD high-dose and low-dose groups,and positive control group.Four weeks after modeling,WTD groups and the positive control group were given WTD(80,20 g·kg^(-1))and glucosamine sulfate capsules(0.29 g·kg^(-1)),respectively,and the sham-operated group and model group received normal saline of the equivalent volume.After continuous intervention for 4 weeks,hemoxylin-eosin(HE)staining was used to observe the morphological changes of cartilage and Mankin scoring system was employed to score the knee cartilage.Western blot was combined with Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)to detect the protein and mRNA levels of vascular endothelial growth factorα(VEGFA),vascular endothelial growth factor receptor 2(VEGFR2),extracellular signal-related kinase 1/2(ERK1/2)and a disintegrin and metalloproteinase with thrombospondin motifs 4(ADAMTS4).Result:The Mankin score in the model group increased as compared with that in the sham-operated group(P<0.01).Compared with the model group,administration groups demonstrated alleviated articular cartilage defect and low Mankin score(P<0.01),but there was no statistical significance in Mankin score between the WTD groups and positive control group.The protein and mRNA levels of VEGFA,VEGFR2,ERK1/2,and ADAMTS4 in the model group were significantly higher than those in the sham-operated group(P<0.01).The protein expression of VEGFA and ERK1/2 was inhibited in each administration group as compared with that in the model group(P<0.01),and the inhibition in the positive control group was stronger than that in the WTD low-dose group(P<0.05)but weaker than that in the WTD high-dose group(P<0.01).Glucosamine Sulfate capsules suppressed t

关 键 词：温经通络汤 膝骨关节炎 血管生成 血管内皮生长因子(VEGF) 细胞外调节蛋白激酶1/2(ERK1/2) 

分 类 号：R2-0[医药卫生—中医学] R22