阳和汤对早期膝骨关节炎模型兔滑膜炎症的影响

Interventional effect of Yanghe Decoction on synovial inflammation in a rabbit model of early knee osteoarthritis

背景:阳和汤是临床治疗膝骨关节炎的有效方药,前期基础研究表明阳和汤防治骨关节炎的机制与其对关节软骨的保护有关,阳和汤对膝骨关节炎早期滑膜炎症的影响尚未明确。目的:探讨阳和汤对早期膝骨关节炎模型兔滑膜炎症的影响,进一步明确阳和汤治疗早期骨关节炎的疗效及作用机制。方法:将24只4月龄新西兰兔随机分为空白对照组、模型组、阳和汤组和阳性对照组,每组6只。除空白对照组外,其余各组予关节腔注射木瓜蛋白酶造模。造模2周后,空白对照组和模型组给予等量生理盐水灌胃,阳和汤组给予6 g/(kg·d)阳和汤灌胃,阳性对照组给予24 mg/(kg·d)塞来昔布药物灌胃,连续给药2周。最后一次给药后4 h,测量膝关节局部皮温、关节周径,Lequesne评分评估膝关节功能;最后一次灌胃结束后第2天,ELISA法检测关节液中白细胞介素1β、肿瘤坏死因子α、环氧化酶2、前列腺素E2水平;苏木精-伊红染色观察滑膜组织形态;免疫组织化学染色观察滑膜组织中白细胞介素1β、肿瘤坏死因子α、环氧化酶2蛋白表达。结果与结论:(1)与空白对照组比较,模型组膝关节周径增宽(P <0.05),局部皮温、Lequesne评分及关节液中白细胞介素1β、肿瘤坏死因子α、环氧化酶2、前列腺素E2水平均升高(P <0.05),滑膜形态学炎性改变明显,滑膜组织中白细胞介素1β、肿瘤坏死因子α、环氧化酶2蛋白表达均升高(P <0.05);(2)与模型组比较,阳和汤组和阳性对照组膝关节周径减小(P <0.05),局部皮温、Lequesne评分及关节液中肿瘤坏死因子α、白细胞介素1β、环氧化酶2、前列腺素E2水平均降低(P <0.05),滑膜炎性改变减轻,滑膜组织中白细胞介素1β、肿瘤坏死因子α、环氧化酶2蛋白表达均降低(P <0.05);(3)结果表明,阳和汤能抑制早期膝骨关节炎模型兔关节液和滑膜组织中肿瘤坏死因子α、白细胞介素1β、环氧化酶2、前列腺素E2的表达,减轻滑膜炎症,改善关节功能,从而延缓膝骨关节炎的进展。

BACKGROUND: Yanghe Decoction is an effective prescription for the clinical treatment of knee osteoarthritis. Our previous basic studies have shown that the mechanism of Yanghe Decoction to prevent and treat knee osteoarthritis is related to the protection of articular cartilage. It is still not unclear about the effect of Yanghe Decoction on early synovial inflammation of knee osteoarthritis. OBJECTIVE: To investigate the effect of Yanghe Decoction on the pain and synovial inflammation in a rabbit model of early knee osteoarthritis, and to further clarify the efficacy and mechanism of Yanghe Decoction in the treatment of early osteoarthritis.METHODS: Twenty-four 4-month-old New Zealand rabbits were randomly divided into blank control group, model group, Yanghe Decoction group and positive control group, with six rabbits in each group. Except for the blank control group, the rabbits in the other groups were injected with papain into the joint cavity for modeling. Two weeks after modeling, the blank control and model groups were given the same amount of normal saline by gavage, the Yanghe Decoction group was given 6 g/kg/d Yanghe Decoction, and the positive control group was given 24 mg/kg/d Celebrex. Administration in each group lasted for 2 weeks. At 4 hours after final administration, the knee joint skin temperature and circumference were measured, and Lequesne MG score was used to evaluate knee joint function. At 2 days after final intragastric administration, ELISA was used to detect the levels of interleukin-1β, tumor necrosis factor-α, cyclooxygenase-2, and prostaglandin E2 in the synovial fluid. Synovial tissue morphology was observed using hematoxylin-eosin staining. Immunohistochemical detection was conducted to detect the expression of interleukin-1β, tumor necrosis factor-α, cyclooxygenase-2 proteins in synovial tissue. RESULTS AND CONCLUSION: Compared with the blank control group, the knee joint circumference in the model group was widened(P < 0.05), and the local skin temperature, Lequesne MG score, and the levels of interleukin-1β, tumor necrosis factor-α, cyclooxygenase-2, and prostaglandin E2 in the synovial fluid were all increased(P < 0.05), synovial membrane morphology and inflammatory changes were obvious, and the expressions of interleukin-1β, tumor necrosis factor-α, cyclooxygenase-2 proteins in synovial tissue were all increased(P < 0.05). Compared with the model group, the knee joint circumference in the Yanghe Decoction group and positive control group decreased(P < 0.05), local skin temperature, Lequesne MG score, and the levels of interleukin-1β, tumor necrosis factor-α, cyclooxygenase-2, and prostaglandin E2 in the synovial fluid were all decreased(P < 0.05), synovial membrane morphology and inflammatory changes were alleviated, and the expressions of interleukin-1β, tumor necrosis factor-α, cyclooxygenase-2 proteins in synovial tissue were all downregulated(P < 0.05). In conclusion, Yanghe Decoction can inhibit the expression of interleukin-1β, tumor necrosis factor-α, cyclooxygenase-2, and prostaglandin E2, reduce synovial inflammation, and improve joint function, thereby delaying the progression of knee osteoarthritis.