摘要
目的探究强筋合剂对骨关节炎大鼠膝关节炎症的影响及其作用机制。方法使用10%木瓜蛋白酶和0.03 mol/L L-半胱氨酸的混合液造模,建立骨关节炎大鼠模型。36只雄性Wistar大鼠随机分为对照组、模型组、强筋合剂低、中、高剂量组、盐酸氨基葡萄糖组,每组6只。其中强筋合剂低、中、高剂量组分别给予蒸馏水制备的体积分数为2.12%、4.35%、8.69%强筋合剂,10 mL/(kg·d),盐酸氨基葡萄糖组给予13 g/L盐酸氨基葡萄糖,10 mL/(kg·d),对照组和模型组给予等量0.9%氯化钠溶液,灌胃治疗4周。治疗期间检测各组大鼠膝关节肿胀度。治疗结束后,采用ELISA法和qPCR法分别检测各组大鼠血清IL-1β水平及膝关节软骨组织中转化生长因子β1(TGF-β1)mRNA相对表达量;采用Western blot法检测各组大鼠膝关节软骨组织中基质金属蛋白酶(MMP)-13、环氧化酶(COX)-2、Ⅱ型胶原蛋白(CollagenⅡ)相对表达水平;采用HE染色法观察各组大鼠膝关节病理形态。结果与对照组大鼠相比,模型组大鼠膝关节骨细胞损害严重,可见大量炎性细胞,膝关节肿胀明显;血清IL-β1水平升高,软骨组织中TGF-β1 mRNA和MMP-13、COX-2蛋白相对表达水平增加,CollagenⅡ蛋白相对表达水平降低,差异均有统计学意义(均P<0.05)。与模型组大鼠相比,强筋合剂高剂量组和盐酸氨基葡萄糖组大鼠膝关节可见少量炎性细胞聚集,膝关节骨面趋于光滑,膝关节肿胀度降低;强筋合剂中、高剂量组与盐酸氨基葡萄糖组大鼠血清中IL-1β水平显著降低;软骨组织中MMP-13、COX-2蛋白相对表达水平下降,CollagenⅡ相对表达水平增加,TGF-β1 mRNA相对表达量降低,差异均有统计学意义(均P<0.05)。结论强筋合剂可通过降低骨关节炎大鼠血液中IL-1β水平,抑制软骨组织中TGF-β1、MMP-13、COX-2蛋白表达和促进软骨组织中CollagenⅡ表达来改善膝关节炎症。
Objective To explore the effect of Qiangjin mixture on joint inflammation of rats with knee osteoarthritis and its mechanism.Methods The rat model of knee osteoarthritis was established with intraarticular injection of 10%papain and 0.03 mol/L L-cysteine in SD rats.The rats in Qiangjin mixture low-dose,medium-dose and high-dose groups were given 2.12%,4.35%and 8.69%of Qiangjin mixture[10 mL/(kg·d)]by gavage for 4 weeks;rats in glucosamine hydrochloride group were given 13 g/L glucosamine hydrochloride[10 mL/(kg·d)]and rats in control group and model group were given equal volume of 0.9%sodium chloride solution.There were 36 male Wister rats,6 in each group.The swelling of knee was observed during the treatment.After treatment,the serum levels of IL-1βand the expression level of TGF-β1 mRNA in knee cartilage were detected by ELISA and qPCR,respectively.The relative expressions of matrix metalloproteinase(MMP)-13,cyclooxygenase(COX)-2 and Collagen II in the knee cartilage were detected by Western blot.HE staining was used to observe the pathological changes of knee joint in each group.Results Compared with the control group,the knee bone tissue in the model group were seriously damaged,with a large number of inflammatory cells and obvious knee swelling.The serum IL-β1 increased,the expression levels of TGF-β1 mRNA and MMP-13 and COX-2 protein in cartilage increased,and the expression levels of CollagenⅡprotein decreased(P<0.05).Compared with model group,less inflammatory cell infiltration was observed in the knee joint of rats in Qiangjin mixture high-dose group and glucosamine hydrochloride group,and the knee joint surface tended to be smooth,and the degree of knee joint swelling was reduced.The serum IL-1βlevels of rats in the medium and high dose of Qiangjin mixture and glucosamine hydrochloride groups was significantly decreased;the expression levels of MMP-13 and COX-2 protein were decreased;the expression of CollagenⅡin the cartilage was increased,and the expression of TGF-β1 was decreased(all P<0.05).Conclusion Qiangjin mixture can alleviate knee inflammation by reducing the serum IL-1βlevel,inhibiting the expression of TGF-β1,MMP-13 and COX-2 protein and increasing CollagenⅡlevel in the cartilage.
作者
刘慧
谢海霞
徐惠平
王建中
LIU Hui;XIE Haixia;XU Huiping;WANG Jianzhong(Preparation Room,Huzhou Hospital of Traditional Chinese Medicine Affiliated to Zhejiang Chinese Medicine University,Huzhou 313000,China)
出处
《浙江医学》
CAS
2023年第14期1492-1497,共6页
Zhejiang Medical Journal
基金
浙江省中医药科技计划青年人才基金项目(2021ZQ083)。
