摘要
目的研究白芍总苷(TGP)通过调节哺乳动物雷帕霉素靶蛋白(mTOR)/缺氧诱导因子-1α(HIF-1α)通路对实验性变态反应性脑脊髓炎(EAE)小鼠免疫功能的影响。方法 C57BL/6J小鼠120只,随机分为对照组(n=30)、模型组(n=30)及低(n=30)、高剂量实验组(n=30)。模型组及低、高剂量实验组小鼠均以髓鞘少突胶质细胞糖蛋白多肽35-55(MOG 35-55)为抗原诱导的方法建立EAE小鼠模型。造模成功后,低、高剂量实验组小鼠分别给予0.2,0.4 g·kg-1·d-1的TGP灌胃,对照组与模型组灌胃等量生理盐水,连续15 d。用流式细胞术检测外周血Treg细胞比例;用蛋白质印迹(Wb)法检测脊髓组织mTOR、HIF-1α蛋白表达;用神经功能评分标准评估小鼠神经功能;用酶联免疫吸附(ELISA)法检测血清白细胞介素-17(IL-17)、白细胞介素-6(IL-6)及肿瘤坏死因子-α(TNF-α)水平。结果对照组、模型组和低、高剂量实验组小鼠外周血Treg细胞比例分别为(5.84±0.37)%,(3.89±0.16)%,(4.73±0.28)%,(5.12±0.34)%;脊髓组织mTOR蛋白相对表达量分别为0.06±0.03,0.92±0.08,0.45±0.13,0.26±0.11;HIF-1α蛋白相对表达量分别为0.14±0.07,0.97±0.03,0.55±0.12,0.32±0.09,差异均有统计学意义(均P<0.05)。结论 TGP可抑制EAE小鼠炎性因子水平,调节免疫功能,改善神经功能,其作用机制可能与抑制mTOR/HIF-1α通路信号转导相关。
Objective To investigate the effects of total glucosides of paeony(TGP) on immune function in mice with experimental allergic encephalomyelitis(EAE) by regulating mammalian rapamycin target protein(mTOR)/hypoxia-inducible factor-1α(HIF-1α) pathway. Methods A total of 120 C57 BL/6 J mice were randomly divided into control group(n=30), model group(n=30) and Exp-L/H groups(n=30). The EAE mouse model was established in model group and Exp-L/H groups using the myelin oligodendrocyte glycoprotein polypeptide 35-55(MOG 35-55) as the antigen-induced method. After successful modeling, Exp-L/H groups were given TGP with 0.2, 0.4 g·kg-1·d-1, control group and model group were given the same amount of normal saline for 15 d. The Treg cell ratio in peripheral blood was detected by flow cytometry;the expression of mTOR and HIF-1α protein in spinal cord tissues were detected by Western blotting(Wb) method;neurological function was assessed using neurological function scoring criteria;serum levels of interleukin-17(IL-17),tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6) were measured by enzyme-linked immunosorbent assay(ELISA). Results The peripheral blood Treg cell ratio of control group,model group and Exp-L/H groups were(5. 84 ± 0. 37) %,(3. 89 ± 0. 16) %,(4. 73 ± 0. 28) %,(5. 12 ± 0. 34) %;the relative expression levels of m TOR protein in the spinal cord tissue were 0. 06 ± 0. 03,0. 92 ± 0. 08,0. 45 ± 0. 13 and 0. 26 ± 0. 11;the relative expression levels of HIF-1αprotein were 0. 14 ± 0. 07,0. 97 ± 0. 03,0. 55 ± 0. 12 and 0. 32 ± 0. 09,all with significant difference(all P < 0. 05).Compared with control group,the neurological function score,serum IL-17,TNF-α,IL-6 levels of model group were significantly increased,Exp-L/H groups were significantly lower than model group,and Exp-H group was lower than Exp-L group(all P < 0. 05). Conclusion TGP can inhibit the levels of inflammatory factors,regulate immune function and improve neurological function in EAE mice,and its mechanism may be related to the inhibition of m TOR/HIF-1α pathway signaling.
作者
杨艳丽
杜杉杉
YANG Yan-li;DU Shan-shan(Basic Medical College,Jining Medical University,Jining 272067,Shandong Province,China;Department of Internal Medicine,First People’s Hospital of Jining,Jining 272000,Shandong Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2020年第24期4025-4027,4034,共4页
The Chinese Journal of Clinical Pharmacology
基金
国家自然青年科学基金资助项目(81601426)
济宁医学院2018年校级教育科学研究基金资助项目(18013)。
关键词
实验性变态反应性脑脊髓炎
白芍总苷
哺乳动物雷帕霉素靶蛋白
缺氧诱导因子-1Α
免疫功能
experimental allergic encephalomyelitis
total glucosides of white paeony
target protein of mammalian rapamycin
hypoxia-inducible factor-1α
immune function
