摘要
目的:从钙调神经磷酸酶/活化T细胞核因子(CaN/NFAT)通路探讨青藤碱(SIN)对胶原诱导性关节炎(CIA)小鼠骨破坏的作用。方法:C57BL/6小鼠60只,随机选取6只为正常组(C组),其余小鼠为造模组。造模组小鼠以牛Ⅱ型胶原(CⅡ)和完全弗氏佐剂的混合乳化剂为抗原尾根部注射,3周后加强免疫,取CⅡ与等体积的不完全弗氏佐剂混合尾根部注射,建立CIA模型。造模成功后,随机分为模型组(M组)、SIN低剂量组(SL组)、SIN高剂量组(SH组)各9只。初次免疫后第31天起,SL组、SH组分别给予SIN 30 mg/kg、300 mg/kg灌胃,连续12 d,每日1次,C组、M组给予等体积的生理盐水。观察小鼠关节肿胀程度并评估关节炎指数(AI);HE染色观察小鼠踝关节病理变化并评价;免疫组化检测CIA小鼠踝关节切片CaNAα、NFAT蛋白表达水平;Western blot检测CIA小鼠滑膜组织CaNAα、NFAT蛋白表达水平。结果:与C组相比,M组关节AI指数、炎症细胞浸润、软骨及骨破坏评分、CaNAα、NFAT蛋白表达均明显升高(P<0.05或P<0.01);与M组相比,SL、SH组的AI指数、炎症细胞浸润、骨及软骨破坏评分、CaNAα、NFAT蛋白水平均明显降低(P<0.05或P<0.01),且呈剂量依赖性。结论:SIN可能通过抑制CaN/NFAT信号通路,抑制破骨细胞分化,改善CIA小鼠骨破坏,为临床治疗类风湿关节炎提供实验依据。
Objective:To investigate the inhibitory effects of sinomenine(SIN)on bone erosion in a collagen-induced arthritis(CIA)mouse model by suppressing the activation of calcineurin(CaN)-nuclear factor of activated T cells/cytoplasmic 1(NFATc1)pathway.Methods:There were 60 C57BL/6 mice,6 of which were randomly selected to normal group,others were ready to be injected with adjuvant.Briefly,bovine native collagenⅡ(CⅡ)was emulsified in equal volumes of complete freund′s adjuvant(CFA).Mice were injected intradermally with the CⅡ/CFA emulsion,using the loose skin at the base of the tail on day 0.On day 21 after the primary immunization,a intradermal booster injection was administered with CⅡ/incomplete Freund′s adjuvant.There were 6 normal mice used as normal control group,and the other 27 successfully established CIA mice were randomly divided into model group(M group),SL group(30 mg/kg)and SH group(300 mg/kg),which were treatment with SIN(n=9).SIN was gavaged to mice at the corresponding dose once daily,while the normal control group and the CIA model group received normal saline,starting on the 31nd day after the first immunization and ending on the 42nd day.Swelling degree and arthritis index(AI)of mice were observed.The histopathological assessments were performed by HE staining.Immunohistochemistry and Western blot were used to detect the expressions of CaNAαand NFATc1 in the synovial and ankle joints.Results:Compared with CIA normal group,AI,scores of inflammation,cartilage damage,bone erosion and levels of NFATcl in the synovium and ankle joints in CIA model groups were significantly increased(P<0.05 or P<0.01).After treatment,compared with the CIA model group,AI,scores of inflammation,cartilage damage,bone erosion and levels of NFATc1 in the synovium and ankle joints(P<0.05 or P<0.01)in all treatment groups were significantly reduced by SIN(P<0.05 or P<0.01).Conclusion:SIN may mitigate bone damage of CIA mice and inhibiting bone erosion by the CaN-NFATc1 pathway.Additionally,our study provides a referab
作者
张浩然
吉金雨
侯晓强
颜岚
梅志刚
冯知涛
ZHANG Hao-Ran;JI Jin-Yu;HOU Xiao-Qiang;YAN Lan;MEI Zhi-Gang;FENG Zhi-Tao(Third-Grade Pharmacological Laboratory on Chinese Medicine Approved by State Administration of Traditional Chinese Medicine,Medical College of China Three Gorges University,Yichang 443002,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2020年第12期1447-1452,共6页
Chinese Journal of Immunology
基金
国家自然科学基金(No.81703783)
湖北省自然科学基金(No.2017CFB126)资助。
