基于NF-κB通路探讨丹参酮ⅡA治疗膝骨关节炎的机制  

Tanshinone Ⅱ alleviates keen osteoarthritis via NF-κB signaling pathway

作　　者：刘江燕 段洪超[2] 张晓峰[2] 徐西林[3] 李朝霞 李俊辰 LIU Jiangyan;DUAN Hongchao;ZHANG Xiaofeng;XU Xilin;LI Chaoxia;LI Junchen(Heilongjiang University of Chinese Medicine,Harbin 150040,Heilongjiang,China;The Second Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150001,Heilongjiang,China;The Third Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150016,Heilongjiang,China)

机构地区：[1]黑龙江中医药大学,黑龙江哈尔滨150040 [2]黑龙江中医药大学第二附属医院,黑龙江哈尔滨150001 [3]黑龙江中医药大学第三附属医院,黑龙江哈尔滨150016

出　　处：《现代中西医结合杂志》2024年第12期1635-1641,1654,共8页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：国家自然科学基金资助项目(81774343);全国名老中医专家传承工作室(2021-2024);黑龙江中医药管理局中医药科研项目(ZHY2023-045);中央支持地方高校改革发展资金人才培养项目(2019年项目)。

摘　　要：目的 基于核因子κB(NF-κB)通路探讨丹参酮ⅡA治疗膝骨关节炎的机制。方法 取52只雄性SD大鼠,随机选择8只作为假手术组,其余大鼠采用改良Hulth法建立骨关节炎模型。将术后2周造模成功的40只大鼠随机分为模型组、玻璃酸钠组及丹参酮ⅡA低、中、高浓度组,每组8只。丹参酮ⅡA各组分别给予每2 mL药液含生药量2.5 mg、5 mg、10 mg的丹参酮ⅡA磺酸钠注射液0.09 mL关节腔注射,模型组和假手术组给予等量生理盐水关节腔注射,玻璃酸钠组给予等量玻璃酸钠关节腔注射,均每周1次。连续干预5周后,ELISA法检测血清中白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)水平;HE染色、番红O固绿染色观察骨关节炎病变程度和胶原纤维含量,Mankin评分量化软骨的退化程度;Western blot法检测膝关节软骨组织中NF-κB p65、p-NF-κB p65、IκBα、Ⅱ型胶原蛋白(ColⅡ)、蛋白聚糖(Aggrecan)蛋白表达情况,qRT-PCR法检测膝关节软骨组织中NF-κB p65、IκBα、ColⅡ、Aggrecan mRNA表达情况。结果 与假手术组比较,模型组大鼠软骨损伤严重,血清IL-6、IL-1β、TNF-α水平及软骨组织中IκBα、ColⅡ、Aggrecan蛋白相对表达量和ColⅡ、Aggrecan mRNA相对表达量均明显降低(P均<0.05),软骨组织中p-NF-κB p65蛋白相对表达量和NF-κB p65、IκBα mRNA相对表达量均明显升高(P均<0.05);与模型组比较,丹参酮ⅡA各组和玻璃酸钠组大鼠软骨损伤减轻,除丹参酮ⅡA低浓度组血清TNF-α水平变化不明显外(P>0.05),丹参酮ⅡA各组和玻璃酸钠组血清IL-6、IL-1β、TNF-α及软骨组织中IκBα、ColⅡ、Aggrecan蛋白相对表达量和ColⅡ、Aggrecan mRNA相对表达量均明显升高(P均<0.05),软骨组织中p-NF-κB p65蛋白相对表达量和NF-κB p65、IκBα mRNA相对表达量均明显降低(P均<0.05)。结论 丹参酮ⅡA可以通过抑制NF-κB信号通路来延缓膝骨关节炎的进展。Objective It is to explore the mechanism of tanshinoneⅡA in the treatment of knee osteoarthritis based on the nuclear factorκB(NF-κB)pathway.Methods Fifty-two male SD rats were taken,in which 8 rats were randomly selected as the sham operation group,and the remaining rats were modeled by modified Hulth method to establish osteoarthritis models.The 40 rats that were successfully modeled in 2 weeks after surgery were randomly divided into model group,sodium hyaluronate group,low,medium and high dose groups of tanshinoneⅡA,with 8 rats in each group.The tanshinoneⅡA groups were given 0.09 mL of sodium tanshinoneⅡA sulfonate injection containing the corresponding dose of raw drug(2.5mg,5mg,10 mg)by joint cavity injection,and the model group and the sham operation group were given equal amount of saline by joint cavity injection,all once per week.After 5 weeks of continuous treatment,the serum levels of interleukin-6(IL-6),interleukin-1β(IL-1β),and tumor necrosis factor-α(TNF-α)were detected by ELISA;the degree of osteoarthritic lesions and the contents of collagen fiber were observed by HE staining and Senna O solid green staining,and the degree of cartilage degradation was quantified by Mankin scores;the protein expressions of NF-κB p65,p-NF-κB p65,IκBαand ColⅡin the cartilage tissues of knee joints were detected by Western blot method,and the mRNA expressions of NF-κB p65,IκBαand ColⅡin the cartilage tissues of knee joints were detected by qRT-PCR method.Results Compared with the sham operation group,the cartilage damage of rats in the model group was severe,and the serum levels of IL-6,IL-1β,TNF-α,relative protein expressions of IκBα,ColⅡ,Aggrecan in cartilage tissues,and relative mRNA expressions of ColⅡ,Aggrecan in cartilage tissues were significantly decreased(all P<0.05),the relative protein expression of p-NF-κB p65 in cartilage tissues,and relative mRNA expressions of NF-κB p65,IκBαin cartilage tissues were significantly increased(all P<0.05).Compared with the model group,

关 键 词：骨关节炎 炎症 软骨基质 NF-ΚB 丹参酮ⅡA 

分 类 号：R-332[医药卫生]