摘要
目的:研究保护素DX(PDX)对类风湿关节炎(RA)大鼠模型的治疗作用及机制,及其对PI3K/AKT/mTOR信号通路的影响。方法:通过皮下注射牛Ⅱ型胶原与弗氏完全佐剂诱导RA大鼠模型,建模后将SD大鼠随机分为4组:对照组(n=12):正常SD大鼠;RA组(n=12):RA模型大鼠;低剂量PDX处理组(n=12,L-PDX组):接受10μg/kg/d PDX治疗的RA模型大鼠;高剂量PDX处理组(n=12,H-PDX):接受20μg/kg/d PDX治疗的RA模型大鼠。各组大鼠治疗4周后,采用ELISA法检测血清中Ig A、Ig G、Ig M、TNF-α、IFN-γ、IL-4和IL-10的水平。通过苏木精伊红(HE)染色评价大鼠踝关节病变。通过免疫组化染色或Western blot检测滑膜组织中PI3K、p-PI3K、AKT、p-AKT、m TOR、p-mTOR、Bcl-2、Bax、LC3-I、LC3-II和Becline-1的表达。结果:与RA组相比,L-PDX组和H-PDX组的关节炎指数(AI)评分均显著降低(P<0.05),炎性细胞浸润、软骨破坏程度及滑膜上皮细胞增生减轻。与RA组相比,L-PDX组和H-PDX组的血清Ig A、Ig G和Ig M含量均降低(P<0.05)。与RA组相比,L-PDX组和H-PDX组的血清TNF-α和IFN-γ水平均降低,IL-4和IL-10水平均升高(P<0.05)。与RA组相比,L-PDX组和H-PDX组大鼠踝关节滑膜组织中的p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR和Bcl-2的蛋白相对表达量均降低,而Bax、LC3-II/LC3-I和Becline-1的蛋白相对表达量均升高(P<0.05)。结论:本研究表明PDX可有效减轻RA大鼠症状,其机制与调节B淋巴细胞活化和体液免疫、纠正Th1/Th2失衡、抑制PI3K/Akt/mTOR信号通路有关。
Objective: To investigate the therapeutic effect and mechanism of protectin DX(PDX) on rheumatoid arthritis(RA) rat model, and its effect on PI3K/AKT/mTOR signaling pathway. Methods: The RA rat model was induced by subcutaneous injection of bovine type Ⅱ collagen and Freund’s complete adjuvant. After modeling, SD rats were randomly divided into 4 groups: Control group(n=12): normal SD rats, RA group(n=12): RA model rats, low-dose PDX group(L-PDX, n=12): RA model rats treated with 10 μg/kg/d PDX, high-dose PDX group(H-PDX, n=12): RA model rats treated with 20 μg/kg/d PDX. After 4 weeks of treatment, the serum levels of Ig A, Ig G, Ig M, TNF-α, IFN-γ, IL-4 and IL-10 were detected by ELISA. Rat ankle joint lesions were evaluated by hematoxylin and eosin(HE) staining. The expressions of PI3K, p-PI3K, AKT, p-AKT, m TOR, p-mTOR, Bcl-2, Bax, LC3-I, LC3-II and Becline-1 in synovial tissue were detected by immunohistochemical staining or Western blot. Results: Compared with RA group, the arthritis index(AI)scores of L-PDX group and H-PDX group were significantly decreased(P<0.05), and the infiltration of inflammatory cells, the degree of cartilage destruction and the proliferation of synovial epithelial cells were reduced. Compared with RA group, the serum Ig A, Ig G and Ig M contents in L-PDX group and H-PDX group were all decreased(P<0.05). Compared with RA group, the serum levels of TNF-α and IFN-γ in L-PDX group and H-PDX group were decreased, and the levels of IL-4 and IL-10 were increased(P<0.05). Compared with RA group, the relative protein expressions of p-PI3K/PI3K, p-AKT/AKT, p-mTOR/mTOR and Bcl-2 in the synovial tissue of the ankle joint of the rats in L-PDX group and H-PDX group were decreased, while the relative protein expressions of Bax, LC3-II/LC3-I and Becline-1were increased(P<0.05). Conclusion: This study shows that PDX can effectively alleviate the symptoms of RA rats, and its mechanism is related to regulating B lymphocyte activation and humoral immunity, correcting Th1/Th2 imbalance, and in
作者
庞琳烜
李颖
李治琴
杜望磊
谢荣华
丁进
PANG Lin-xuan;LI Ying;LI Zhi-qin;DU Wang-lei;XIE Rong-hua;DING Jin(Department of Clinical Immunology,Air Force Medical University,Xi'an,Shaanxi,710032,China;Department of Rheumatology,The First People's Hospital of Xianyang,Xianyang,Shaanxi,712000,China)
出处
《现代生物医学进展》
CAS
2023年第3期401-406,共6页
Progress in Modern Biomedicine
基金
国家自然科学基金项目(81701617)。
