摘要
目的观察黄芪鳖甲丸对膝关节骨性关节炎(KOA)大鼠关节软骨的相关病理改变及相关蛋白在缺氧诱导因子1α(HIF-1α信号通路的表达。方法选取60只SD大鼠随机分为假手术组(10只)和造模组(50只)。假手术组保留大鼠卵巢,造模组在切除大鼠卵巢8周后进行关节腔注射碘乙酸钠1次,然后将造模组分为模型组、塞来昔布组、黄芪鳖甲丸高剂量组、黄芪鳖甲丸中剂量组及黄芪鳖甲丸低剂量组,每组10只。各给药组大鼠药物持续干预4周。在末次给药后检测大鼠血清雌二醇(E2)水平、关节灌洗液的肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)含量,并测量大鼠膝关节横径,观察软骨的外观及病理变化,使用蛋白质免疫印迹法(Western blot)检测各组大鼠软骨的蛋白表达。结果黄芪鳖甲丸各剂量组大鼠膝关节横径、关节灌洗液IL-1β、TNF-α水平均显著少于模型组(P<0.05);黄芪鳖甲丸高、中剂量组大鼠血清E2水平高于模型组(P<0.05)。大体观察黄芪鳖甲丸高剂量组大鼠膝关节软骨表面较模型组光滑,且伴有光泽,缺损不明显;中剂量组大鼠膝关节软骨的中部稍有损伤,有血管增生。镜下观察黄芪鳖甲丸高、中剂量组大鼠膝关节软骨表层光滑,软骨细胞排列规整,细胞形态未见异常,潮线完整。黄芪鳖甲丸各剂量组大鼠软骨光镜软骨检测评价标准(Mankin’s)评分、软骨HIF-1α及基质金属蛋白酶-1(MMP-1)黄染颗粒光密度值(IOD)、软骨HIF-1α、血管内皮生长因子(VEGF)蛋白表达量均显著低于模型组(P<0.05),黄芪鳖甲丸高剂量组大鼠软骨中葡萄糖载体-1(Glut-1)、MMP-1蛋白表达量低于模型组(P<0.05)。结论黄芪鳖甲丸降低软骨组织损伤及炎症程度,其作用可能与HIF-1α调控Glut-1、VEGF、MMP-1的表达有关。
Objective To explore the effect of Huangqibiejia pills in the treatment of knee osteoarthritis(KOA)by observing the changes of articular cartilage,and the expression of related proteins in hypoxia-inducible factor 1α(HIF-1α)signaling pathway.Methods 60 SD rats were randomly divided into the sham group(10 rats)and model group(50 rats).The ovaries of the rats in the sham group were kept,while the rats in the model group were injected with sodium iodoacetate once in the knee joint cavity 8 weeks after ovary resection.Then the rats were divided into KOA-model group,celecoxib group and Huangqibiejia pills high-dose group,medium-dose group and lowdose group,with 10 rats in each group.The rats in each group were given drug intervention for 4 weeks.After the last administration,the serum estradiol(E2)level,tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)contents of the lavage fluid were detected,and the transverse diameter of the knee joint was measured to observe the appearance and pathological changes of cartilage.The protein expression of cartilage in each group was detected by Western blot.Results The contents of IL-1βand TNF-αin the transverse diameter of knee joint and in the irrigation solution in each dose groups of Huangqibiejia pills were significantly lower than those in the KOA-model group(P<0.05).The level of serum E2in Huangqibiejia pills high-dose group and Huangqibiejia pills medium-dose group was higher than that in KOA-model group(P<0.05).The surface of the knee cartilage in Huangqibiejia pills high-dose group was smoother than that in KOA-model group,and the defects were not obvious.The middle part of the knee cartilage was slightly damaged and there was vascular hyperplasia in the Huangqibiejia pills medium-dose group.Under microscope,the knee cartilage surface of rats in Huangqibiejia pills high-dose group and Huangqibiejia pills medium-dose group was smooth,the chondrocytes were arranged neatly,the cell morphology was not abnormal,and the tidal line was complete.Compared with the KOA-model group,the Mankin’s score,the integrated optical density(IOD)value of HIF-1αand matrix metalloproteinase-1(MMP-1)yellow dye granules,the expression of HIF-1αand vascular endothelial growth factor(VEGF)protein in each dose groups of Huangqibiejia pills were significantly lower than those in the KOA-model group(P<0.05).The expression levels of glucose transport-1(Glut-1)and MMP-1 in cartilage of rats in high dose group were lower than those in KOA-model group(P<0.05).Conclusion Huangqibiejia pills can reduce the degree of cartilage injury and inflammation in KOA model rats,and the mechanism is related to the regulation of Glut-1,VEGF and MMP-1 expression by HIF-1α.
作者
谭志韵
罗汉文
彭莎
赵新新
赖亦静
TAN Zhiyun;LUO Hanwen;PENG Sha;ZHAO Xinxin;LAI Yijing(Department of Orthopaedics,Foshan Hospital of Traditional Chinese Medicine,Foshan,528000,Guangdong,China;The Fifth Clinical Medical College of Guangzhou University of Chinese Medicine,Guangzhou,510405,Guangdong,China)
出处
《大医生》
2023年第5期9-14,共6页
Doctor
基金
佛山市自筹经费类科技计划项目(医学类科技攻关)(编号:1920001001420)。
关键词
膝关节骨性关节炎
黄芪鳖甲丸
缺氧诱导因子
软骨
Knee osteoarthritis
Huangqibiejia pills
Hypoxia-inducible factor 1α
Cartilage
作者简介
谭志韵,硕士研究生,副主任中医师,研究方向:中医骨伤科学。
