乌头汤对膝骨关节炎大鼠模型的治疗作用

Therapeutic effects of aconitum carmichaelii decoction on a rat model of knee osteoarthritis

目的探讨乌头汤基于核因子E2相关因子2(nuclear factor E2 related factor 2,Nrf2)/血红素加氧酶-1(heme oxygenase-1,HO-1)通路对膝骨关节炎(knee osteoarthritis,KOA)大鼠模型的治疗作用。方法通过向膝关节腔内注射木瓜蛋白酶溶液建立KOA大鼠模型;将造模后的大鼠随机分为KOA组、乌头汤组(4.2 g/kg乌头汤)、抑制剂组(30 mg/kg ML385)、乌头汤+抑制剂组(4.2 g/kg乌头汤+30 mg/kg ML385),每组10只,另取10只大鼠腔内注射生理盐水作为对照组。采用苏木精-伊红染色法观察软骨组织病理学变化并进行损伤评分;采用ELISA法检测血清中炎症因子[肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)/白细胞介素-1β(interleukin-1β,IL-1β)]及氧化应激[丙二醛(malondialdehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)]指标;采用qRT-PCR法及Western blotting法检测Nrf2、HO-1表达。结果与对照组相比,KOA组软骨表面粗糙不平,细胞排列紊乱,层状结构消失,IL-1β、TNF-α、MDA含量增加,SOD、Nrf2、HO-1 mRNA及蛋白表达降低,差异有统计学意义(P<0.05);与KOA组相比,乌头汤组病理损伤得到缓解,IL-1β、TNF-α、MDA含量降低,SOD、Nrf2、HO-1 mRNA及蛋白表达增加,差异有统计学意义(P<0.05),但抑制剂组病理损伤进一步加剧,IL-1β、TNF-α、MDA含量增加,SOD、Nrf2、HO-1 mRNA及蛋白表达降低,差异有统计学意义(P<0.05);与乌头汤组相比,乌头汤+抑制剂组病理损伤较为严重,IL-1β、TNF-α、MDA含量增加,SOD、Nrf2、HO-1 mRNA及蛋白表达降低,差异有统计学意义(P<0.05);与抑制剂组相比,乌头汤+抑制剂组病理损伤稍有改善,IL-1β、TNF-α、MDA含量降低,SOD、Nrf2、HO-1 mRNA及蛋白表达增加,差异有统计学意义(P<0.05)。结论乌头汤可通过激活Nrf2/HO-1通路对KOA大鼠起治疗作用。

Objective To investigate the therapeutic effects of aconitum carmichaelii decoction based on the nuclear factor E2 related factor 2(Nrf2)/heme oxygenase-1(HO-1)pathway on a rat model of knee osteoarthritis(KOA).Methods KOA rat model was established by injecting papain solution into the knee joint;the rats after modeling were randomly grouped into KOA group,aconitum carmichaelii decoction group(4.2 g/kg aconitum carmichaelii decoction),inhibitor group(30 mg/kg ML385),aconitum carmichaelii decoction+inhibitor group(4.2 g/kg aconitum carmichaelii decoction+30 mg/kg ML385),with 10 rats in each group,and another 10 rats were injected intracavitally with physiological saline as the control group.Hematoxylin-eosin staining was used to observe the histopathological changes of cartilage in cartilage and to score the damage;serum indicators of inflammatory factor[tumor necrosis factor-α(TNF-α)/interleukin-1β(IL-1β)]and oxidative stress[malondialdehyde(MDA)and superoxide dismutase(SOD)]were measured by ELISA;Nrf2 and HO-1 expressions were detected by qRT-PCR and Western blotting.Results Compared with the control group,the cartilage surface of KOA group was rough and uneven,disordered cell arrangement,and disappearance of lamellar structure,increased content of IL-1β,TNF-αand MDA,and decreased expressions of SOD,Nrf2,and HO-1 mRNA and protein(P<0.05);compared with the KOA group,the pathological damage of aconitum carmichaelii decoction group was relieved,with decreased contents of IL-1β,TNF-α,and MDA,and increased expressions of SOD,Nrf2,and HO-1 mRNA and protein(P<0.05);however,the pathological damage in the inhibitor group was further aggravated,with increased content of IL-1β,TNF-α,and MDA,and decreased expressions of SOD,Nrf2,and HO-1 mRNA and protein(P<0.05);compared with the aconitum carmichaelii decoction group,aconitum carmichaelii decoction+inhibitor group had more serious pathological damage,with increased content of IL-1β,TNF-α,and MDA,and decreased expressions of SOD,Nrf2,and HO-1 mRNA and protein(P<0.05);compared with the inhibitor group,the pathological damage in the aconitum carmichaelii decoction+inhibitor group was slightly improved,with decreased content of IL-1β,TNF-α,and MDA,and increased expressions of SOD,Nrf2,and HO-1 mRNA and protein(P<0.05).Conclusion Aconitum carmichaelii decoction can treat KOA rats by activating Nrf2/HO-1 pathway.