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目的探讨中药通痹灵(TBL)抗软骨破坏的作用机制。方法以胶原诱导型关节炎(CIA)大鼠为动物模型,分组灌胃治疗后,做大鼠趾关节病理切片,常规HE染色,观察软骨病变;培养软骨细胞,以鼠重组白细胞介素-1β(rrIL-1β)体外刺激软骨细胞导致软骨细胞基质降解作为药理模型,加用不同浓度的通痹灵总碱,并以地塞米松作为对照组,培养48h后,收集细胞上清液,用阿利新蓝法检测葡糖胺聚糖(GAG)含量;用硝酸酶还原法检测NO的含量。结果①趾关节病理HE切片评价结果显示:通痹灵总碱组软骨破坏程度小于造模组(P<0.05〉;②不同浓度的rrIL-1β(1、10、100、500、1000ng/ml)促进了软骨细胞蛋白多糖的降解(P<0.01),且呈量效关系;③不同浓度的通痹灵总碱(200、100、50、25mg/L)及地塞米松可明显地抑制软骨细胞蛋白多糖的降解(P<0.01)及NO的产生(P<0.01)。结论通痹灵总碱可抑制CIA大鼠趾关节软骨破坏;体外实验结果显示通痹灵总碱可对抗软骨细胞蛋白多糖的降解,其作用途径可能是通过抑制NO的生成来实现的。
Objective To investigate the mechanism of the anti-cartilage destruction of traditional Chinese medicine Tongbiling (TBL). Methods The rat model of collagen-induced arthritis (CIA) was used as an animal model. After group-sparing, pathological sections of rat toe joints were made and routinely stained with HE to observe cartilage lesions. Chondrocytes were cultured to reconstitute interleukin-1β in rats. (rrIL-1β) In vitro stimulation of chondrocytes leads to chondrocyte matrix degradation. As a pharmacological model, addition of different concentrations of TBA, and dexamethasone as a control group, after 48 hours of culture, the cell supernatant was collected, using Ali New blue method was used to detect the content of glycosaminoglycan (GAG); the content of NO was detected by nitric acid enzyme reduction method. Results 1The results of HE slice evaluation of the toe joint pathology showed that the degree of cartilage destruction in Tongzhuling total alkali group was less than that in the model group (P<0.05>; 2 different concentrations of rrIL-1β (1, 10, 100, 500, 1000 ng/ml). The degradation of proteoglycans in chondrocytes was promoted (P<0.01) in a dose-effect relationship; (3) Different concentrations of Tongluojing total base (200, 100, 50, 25 mg/L) and dexamethasone significantly inhibited chondrocytes Degradation of proteoglycans (P<0.01) and production of NO (P<0.01).Conclusions Tongkeling total base can inhibit cartilage destruction of CPA rat toe joints; in vitro experiments show that Tongzhuling total base can fight chondrocyte proteoglycans The degradation may be effected by inhibiting the production of NO.