乌头汤对膝骨关节炎大鼠模型的治疗作用

doi:10.6040/j.issn.1671-7554.0.2023.0950

摘要/Abstract

摘要： 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。

Abstract: 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-α and 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.

Key words: Aconitum carmichaelii decoction, Nuclear factor E2 related factor 2/Heme oxygenase-1 pathway, Knee osteoarthritis, Therapeutic mechanism

中图分类号:

R274

王莉莎,王上增,史栋梁,张仲博,任博文,王云飞,郭中华,周晓宁. 乌头汤对膝骨关节炎大鼠模型的治疗作用[J]. 山东大学学报 (医学版), 2024, 62(5): 64-71.

WANG Lisha, WANG Shangzeng, SHI Dongliang, ZHANG Zhongbo, REN Bowen, WANG Yunfei, GUO Zhonghua, ZHOU Xiaoning. Therapeutic effects of aconitum carmichaelii decoction on a rat model of knee osteoarthritis[J]. Journal of Shandong University (Health Sciences), 2024, 62(5): 64-71.

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