骨灵膏及其拆方制剂对骨关节炎氧化损伤及低氧诱导因子的作用  

Effect of Gulinggao and its Components on Oxidative Injury and Hypoxia Inducible Factors in Osteoarthritis

作　　者：陈文超[1,2] 周然[2] 王永辉[2] 李艳彦[2] 柴智[2] 高丽[2] 

机构地区：[1]湖北中医药大学,湖北武汉430065 [2]山西中医学院,山西太原030024

出　　处：《世界中西医结合杂志》2014年第6期651-655,共5页World Journal of Integrated Traditional and Western Medicine

基　　金：山西省科技厅科技创新重点团队建设项目(No.2012081018)

摘　　要：目的通过骨灵膏及其拆方制剂对骨关节炎超氧化物歧化酶(SOD)、丙二醛(MDA)、脂质过氧化物(LPO)及低氧诱导因子-1α(HIF-1α)、低氧诱导因子-2α(HIF-2α)、血管内皮生长因子(VEGF)的影响,探讨骨灵膏及其拆方制剂抗骨关节炎(OA)软骨损伤的作用机制。方法 60只雌性SD大鼠随机分为正常组、模型组、骨灵膏组、骨膏组、灵膏组及塞来昔布组6组,每组10只,除正常组外其余各组采用冷固法6周建立OA模型,造模成功后除正常组和模型组给予生理盐水灌胃外,其余组分别给予骨灵膏、骨膏、灵膏及塞来昔布灌胃,给药8周后,取大鼠血清、膝关节软骨组织,采用ELISA检测血清SOD、MDA、LPO的含量,免疫荧光法检测HIF-1α、HIF-2α、VEGF的阳性表达,FQ-RT-PCR检测HIF-1α、HIF-2α、VEGF mRNA的表达。结果模型组与正常组及骨灵膏组比较能显著增加血清MDA、LPO的含量(P<0.01),增加SOD的含量(P<0.01);骨灵膏组与骨膏及灵膏比较能增加SOD的含量(P<0.01),降低MDA、LPO的含量(P<0.01);模型组与正常组比较能显著增加软骨HIF-1α、HIF-2α、VEGF的阳性率(P<0.01),骨灵膏组与模型组、骨膏组、灵膏组、塞来昔布组比较能降低HIF-1α、HIF-2α、VEGF的阳性率(P<0.01);模型组与正常组比较能显著增加软骨HIF-1α、HIF-2α、VEGF mRNA的表达(P<0.01),骨灵膏组与模型组、骨膏组、灵膏组、塞来昔布组比较能降低HIF-1α、HIF-2α、VEGF mRNA的表达(P<0.01)。结论骨灵膏可能通过增加血清SOD含量,降低LPO、MDA的含量,纠正OA氧代谢失衡对软骨组织的氧化损伤,抑制组织HIF-1α、HIF-2αmRNA的表达,从而抑制异常低氧环境HIF-1α向HIF-2α的转化,降低其下游靶基因VEGF mRNA在软骨组织的表达,减轻了关节骨赘的形成,对抗OA软骨损伤,其作用明显优于骨膏、灵膏、塞来昔布。Objective To explore the effect mechanism of Gulinggao and its components on carti-lage injury in osteoarthritis（ OA）by their impacts on SOD,MDA,LPO,HIF - 1α,HIF - 2α and VEGF. Methods Sixty female SD rats were randomized into a normal group,a model group,a Gulinggao group,a Gugao group,a Linggao group and a Celecoxib group,10 rats in each one. Except in the normal group,the fro-zen method was used to prepare OA model in each of the rest groups. After successfully modeling,the gastric infusion was applied in the normal and model groups,Gulinggao,Gugao,Linggao and Celecoxib were used for gastric infusion in the rest groups separately. In 8 weeks of medication,the serum and cartilage tissue of knee joint were collected from the rats. ELISA was adopted to determine the levels of SOD,MDA and LPO. Immu-nofluorescence method was used to determine the positive expressions of HIF - 1α,HIF - 2α and VEGF. FQ- RT - PCR was applied to determine the expressions of HIF - 1α,HIF - 2α and VEGF mRNA. Results Compared with the model group and Gulinggao group,the levels of MDA and LPO were increased significantly＆amp;nbsp;（P ﹤ 0. 01）and SOD was increased（P ﹤ 0. 01）in the model group. Compared with Gugao group and Linggao group,SOD was increased（P ﹤ 0. 01）and MDA and LPO were decreased in the Gulinggao group（P ﹤ 0. 01）. Compared with the model group,Gugao group,Longgao group and Celecoxib group,the positive rates of HIF- 1α,HIF - 2α and VEGF were reduced in the Gulinggao group（P ﹤ 0. 01）. Compared with the normal group,the expressions of HIF - 1α,HIF - 2α and VEGF mRNA were increased significantly in the model group（P ﹤ 0. 01）. Compared with the model group,Gugao group,Linggao group and Celecoxib group,HIF -1α,HIF - 2α and VEGF mRNA expressions were decreased in Gulinggao group（P ﹤ 0. 01）. Conclusion Guolinggao increases SOD level,reduces LPO and MDA levels,rectifies OA oxidative metabolic imbalance in oxidative injury of cartilage tissue and inhibits the expressions of HIF

关 键 词：骨关节炎 低氧诱导因子-1Α 低氧诱导因子-2α 血管内皮生长因子 

分 类 号：R285.5[医药卫生—中药学]