中药单体姜黄素缓解大鼠骨关节炎的分子机制

Molecular mechanism of traditional Chinese medicine monomer curcumin in alleviating osteoarthritis in rats

目的探讨了中药单体姜黄素缓解大鼠骨关节炎的分子机制。方法30只SD大鼠随机数字表法分为对照组、关节炎组和姜黄素组,每组10只。关节炎组和姜黄素组大鼠采用木瓜蛋白酶局部注射复制骨关节炎动物模型。姜黄素组大鼠经关节腔注射姜黄素50 mg/kg,对照组和关节炎组经关节腔注射等体积生理盐水。连续治疗3周后进行后续研究。测定3组大鼠膝关节最大活动度和关节滑膜厚度;采用分光光度法测定3组大鼠滑膜上清中糖胺多糖的含量;采用Mankin组织学评分分析关节炎病变程度;采用酶联免疫吸附试验(ELISA)分析滑膜液炎性因子水平;采用马松染色分析膝关节胶原纤维含量。组间比较采用单因素方差分析。结果姜黄素组大鼠膝关节最大屈伸角度[(136.87±6.42)°]明显小于关节炎组[(156.81±13.90)°],差异有统计学意义(t=4.714,P<0.05)。姜黄素组大鼠膝关节Mankin评分[(2.36±0.24)分]明显高于关节炎组[(1.50±0.29)分],差异有统计学意义(t=18.051,P<0.05)。姜黄素组大鼠膝关节糖胺聚糖水平[(501.90±26.92)ng/mg]明显高于关节炎组[(445.80±49.22)ng/mg],差异有统计学意义(t=3.162,P<0.05)。姜黄素组大鼠膝关节TNF-α和IL-1β水平[(143.33±20.58)、(41.20±5.98)ng/mg]明显低于关节炎组膝关节TNF-α和IL-1β水平[(255.28±51.48)、(101.03±14.21)ng/mg],差异有统计学意义(t=6.386、12.272,P<0.05)。姜黄素组膝关节p-p65蛋白表达水平(1.49±0.13)明显低于关节炎组膝关节p-p65蛋白表达水平(2.36±0.37),差异有统计学意义(t=7.063,P<0.05)。结论姜黄素可显著下调关节炎炎症水平,促进受损关节胶原纤维合成,缓解骨关节炎的症状。

Objective To explore the molecular mechanism of traditional Chinese medicine monomer curcumin in alleviating osteoarthritis in rats.Methods Totally,30 SD rats were randomly divided into control group,arthritis group and curcumin group,with 10 rats in each group by random number table method.Rats in arthritis group and curcumin group were injected with papain to replicate the animal model of osteoarthritis.The animals in curcumin group were injected with 50 mg/kg curcumin through the articular cavity,and those in control group and arthritis group were injected with equal volume of normal saline through the articular cavity.The follow-up study was carried out after 3 weeks of continuous treatment.The maximum range of motion and synovial thickness of the knee joint in the three groups were measured.The content of glycosaminoglycan in the synovial supernatant in the three groups was determined by spectrophotometry.The degree of arthritis was analyzed by Mankin histological score.The levels of inflammatory factors in synovial fluid were analyzed by enzyme linked immunosorbent assay(ELISA).Masson staining was used to analyze the content of collagen fibers in knee joint.One way ANOVA was used for comparison between groups.Results The maximum knee flexion and extension angle[(136.87±6.42)°]in curcumin group were significantly lower than those in arthritis group[(156.81±13.90)°,t=4.714,P<0.05].The Mankin score of knee joint in curcumin group(2.36±0.24)was significantly higher than that in arthritis group(1.50±0.29,t=18.051,P<0.05).The glycosaminoglycan level in curcumin group[(501.90±26.92)ng/mg]was significantly higher than that in arthritis group[(445.80±49.22)ng/mg,t=3.162,P<0.05].Tumor necrosis factor-α(TNF-α)and interleukin(IL)-1βin knee joint of rats in curcumin group[(143.33±20.58),(41.20±5.98)ng/mg]were significantly lower than those in arthritis group[(255.28±51.48),(101.03±14.21)ng/mg,t=6.386,12.272,P<0.05].The expression level of p-p65 protein in curcumin group(1.49±0.13)was significantly lower than that in arthritis group(2.36±0.37,t=7.063,P<0.05).Conclusion Curcumin can significantly reduce the level of arthritis,promote the synthesis of collagen fibers in damaged joints and alleviate the symptoms of osteoarthritis.