温经通络汤对小鼠膝骨关节炎软骨形态改变和VEGF、MMP-13、HIF-1α表达情况的影响  

Effects of Wenjing Tongluo Decoction on Changes of Cartilage Morphology and Expressions of VEGF,MMP-13 and HIF-1αin Mice with KOA

作　　者：许炜民 钱佳佳[1] 韩龙 许奇 黄桂成[1,2] XU Weimin;QIAN Jiajia;HAN Long;XU Qi;HUANG Guicheng(Nanjing University of Chinese Medicine,Nanjing 210023,China;Jiangsu Province Hospital of Chinese Medicine Affiliated to Nanjing University of Chinese Medicine)

机构地区：[1]南京中医药大学,江苏南京210023 [2]南京中医药大学附属江苏省中医院,江苏南京210023

出　　处：《中医药信息》2020年第6期6-12,共7页Information on Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81774341)。

摘　　要：目的:观察温经通络汤对小鼠膝骨关节炎模型核血管内皮生长因子(VEGF),基质金属蛋白酶-13(MMP-13),缺氧诱导因子-1α(HIF-1α)的影响,探讨其对膝骨关节炎的作用机制。方法:采用前交叉韧带离断(ACLT)法建立小鼠膝骨关节炎模型,将30只C57BL/6小鼠随机分为假手术组(A组)、模型组(B组)、温经通络汤高剂量组(C组)、温经通络汤低剂量组(D组)、阳性药组(E组)。造模成功后A组与B组分别予以10 mL/(kg·d)生理盐水灌胃,C组与D组分别予以10 mL/(kg·d)温经通络汤灌胃,E组予以10 mL/(kg·d)美洛昔康水溶液灌胃,持续2周。采用组织形态学观察及Mankin's评分评估各组小鼠膝关节表面软骨的损伤程度。采用免疫组化法观察各组膝关节表面软骨中VEGF、MMP-13和HIF-1α的表达水平,采用酶联免疫吸附检测各组膝关节软骨组织中血管内皮生长因子(VEGF)、基质金属蛋白酶的表达水平。结果:组织形态学观察及Mankin's评分显示与假手术组相比,其余各组的评分均明显升高(P<0.05),与模型组相比,高剂量组、低剂量组、阳性药组的评分明显下降(P<0.05)。免疫组化结果表明,高剂量组、低剂量组和阳性药组中小鼠膝关节软骨的VEGF、MMP-13、HIF-1α表达相较于模型组均明显下降(P<0.05)。ELISA检测结果显示,与假手术组相比,其余各组的小鼠血清中VEGF、MMP-13及HIF-1α表达均有明显升高(P<0.05),其中模型组小鼠血清中VEGF、MMP-13及HIF-1α的表达最高,高剂量组、低剂量组和阳性药组与模型组相比均有所下降(P<0.05)。结论:温经通络汤可能通过调控HIF-1α抑制VEGF及MMP-13的表达发挥缓解及治疗膝骨关节炎的作用。Objective:To observe the effects of Wenjing Tongluo Decoction(WJTLT)on vascular endothelial growth factor(VEGF),matrix metalloproteinase-13(MMP-13)and hypoxia-induced factor-1α(HIF-1α)in mice with knee osteoarthritis(KOA),thus to explore its action mechanism in the treatment of KOA.Methods:30 C57BL/6 mice were randomly divided into the sham operation group(group A),the model group(group B),the high-dose group(group C),the-low dose group(group D)and the positive drug group(group E).After successful modeling,group A and group B received normal saline gavage 10 ml/(kg·d),group C and group D were treated with different doses of WJTLT 10 ml/(kg·d),and group E was given Meloxicam aqueous solution 10 ml/(kg·d).The treatment duration was two weeks.Histological observation and Mankin's scores were used to evaluate the damage of the surface cartilage of the knee joint.Immunohistochemistry was used to observe the expression levels of VEGF,MMP-13 and HIF-1αin the surface cartilage of the knee,and the expression levels of vascular endothelial growth factor(VEGF)and matrix metalloproteinase in the cartilage were detected by EILSA method.Results:Histopathological observation and Mankin's scores showed that compared to group A,the scores of other groups were significantly increased(P<0.05);compared to group B,the scores of group C,group D and group E were significantly decreased(P<0.05).Immunohistochemistry showed that the expressions of VEGF,MMP-13 and HIF1αin knee cartilage were significantly decreased in group C,D and E compared to those in group B(P<0.05).ELISA results showed that,compared to group A,the serum expressions of VEGF,MMP-13 and HIF-1αwere significantly increased in other groups(P<0.05),among which,the expressions of VEGF,MMP-13 and HIF-1αwere the highest in group B;and they were all decreased in group C,D and E compared to group B(P<0.05).Conclusion:WJTLT may play a role in treating KOA by regulating HIF-1αand inhibiting the expressions of VEGF and MMP-13.

关 键 词：温经通络汤 膝骨关节炎 血管内皮生长因子 基质金属蛋白酶13 缺氧诱导因子1Α 

分 类 号：R285.5[医药卫生—中药学]