化湿补肾法对膝骨关节炎大鼠软骨修复机制的研究

Mechanism of dampness-resolving kidney-tonifying method in cartilage repair in rats with knee osteoarthritis

目的探讨化湿补肾法治疗膝骨关节炎(KOA)的机制。方法将48只SPF级SD大鼠随机分为空白组、模型组、化湿定痛汤组、龟鹿二仙胶组、化湿补肾法组,采用木瓜蛋白酶关节腔注射建立膝骨关节炎模型,造模成功后各组大鼠灌胃相应药物,化湿定痛汤组灌胃化湿定痛汤4.68 g/(kg·d),龟鹿二仙胶组灌胃龟鹿二仙胶汤2.7 g/(kg·d),化湿补肾法组灌胃化湿补肾法中药7.38 g/(kg·d),连续灌胃28 d,于第28天对大鼠进行取材,取右侧股骨行HE染色观察软骨形态,取右侧胫骨平台滑膜、关节液用ELISA检测前列腺素-2(PGE-2)、环氧化和酶-2(COX-2)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)的表达,取右胫骨平台软骨用qPCR检测p38、p53、Ⅱ型胶原的表达。结果 HE染色形态结果表明化湿补肾法组、龟鹿二仙胶组、化湿定痛汤组均优于模型组。与龟鹿二仙胶组相比,化湿补肾法组PGE-2、COX-2、IL-1β、TNF-α表达量下降(P<0.05),软骨的p38、p53表达降低(P<0.05),Ⅱ型胶原表达升高(P<0.05)。结论化湿补肾法可有效减轻滑膜和关节液中炎症因子表达,增加软骨细胞合成代谢,抑制软骨细胞凋亡,从而促进关节软骨的修复,延缓KOA的进程。

Objective To preliminarily explore the mechanism of treating knee osteoarthritis(KOA) by dampness-resolving kidney-tonifying method. Methods 40 SPF SD rats were randomly divided into blank group, model group, dampness-resolving pain-relieving decoction(DP) group, Guilu Erxian(GE) group, and dampness-resolving kidney-tonifying(DK) group(n=8). The model of osteoarthritis was established by injecting papain into the articular dise. After successfully establishing the model, rats in the DP group, GE group and DK group were gavaged with DP decoction 4.68 g/(kg·d), GE decoction 2.7 g/(kg·d) and DK decoction 7.38 g/(kg·d) respectively for 28 days in succession, and the blank group and model group were gavaged with an equal amount of normal saline solution. After 28 days, the cartilage of right femur was taken for morphologic observation with HE staining. The expression of PGE, COX2, IL-1β and TNF-α in synovium and synovial fluid of right tibial plateau were observed with ELISA. The expression of p38, p53 and type Ⅱ collagen of right tibial plateau were detected with PCR. Results The pathological morphologic changes of articular cartilage observed with HE staining showed better results in the DK group, GE group and DP group compared with the model group. The expression of PGE-2, COX-2 and IL-1 beta were decreased in various degrees in the DK group compared with the GE group(P<0.05). In the DK group, the expression of p38 and p53 in the cartilage was decreased, while the expression of type Ⅱ collagen was increased compared with the GE group(P<0.05). Conclusion Dampness-resolving kidney-tonifying method seems to effectively reduce the expression of inflammatory factors in synovium and synovial fluid, promote the anabolism of chondrocytes, and inhibit the apoptosis of chondrocytes, thus promoting the repair of articular cartilage, and delaying the process of KOA.