摘要
目的:探讨内吗啡肽-1(EM-1)对高糖诱导的人外周血来源树突状细胞(PBDC)成熟表型、细胞因子分泌、T细胞增殖和TLR4表达的影响,探讨EM-1对DC免疫功能的调节机制。方法:本研究外周血样品取自18-20岁学生志愿者,将外周血单个核细胞(PBMNC)诱导成未成熟的树突状细胞(imDC)。使用高浓度葡萄糖作为刺激因素,EM-1作为干预因素,将实验分为正常葡萄糖(NG)、高糖(HG)、加入EM-1的高葡萄糖(EM)、加入EM-1和纳洛酮的高葡萄糖(Nal)共5组。应用流式细胞术检测PBDC表型变化,ELISA检测PBDC与自体淋巴细胞共培养分泌的细胞因子的变化,CFSE法检测T淋巴细胞的增殖情况,RT-PCR法检测PBDC表面TLR4的表达。结果:与HG组比较,EM组PBDC表面分子CD86、CCR7和CD36表达上调(P<0.01),而CD83的表达无统计学意义。EM组PBDC分泌的IL-12,IL-10和PBDCs刺激的T淋巴细胞增殖指数均下降。与EM组比较,Nal组CD86,CCR7,CD36表达降低(P<0.01),而CD83表达几乎没有变化(P>0.05)。Nal组T淋巴细胞增殖指数极显著升高(P<0.01)。HG组TLR4灰度比值高于NG组,EM组明显低于HG组(P<0.01)。结果表明,高葡萄糖可以促进PBDC TLR4的表达,而EM-1抑制TLR4的表达。结论:EM-1上调高糖诱导的PBDC表面分子CD86,CCR7和CD36的表达,抑制高糖诱导的P BDC成熟。分泌的炎性细胞因子IL-12和IL-10抑制来自PBDC的T淋巴细胞的增殖,而纳洛酮抑制EM-1的作用。EM-1抑制高糖诱导的PBDC表面TLR4的表达。
Objective： To investigate the effects of endomorphin-1（ EM-1） on the maturation phenotype,cytokine secretion,T cell proliferation and TLR4 expression in human peripheral blood dendritic cells（ PBDCs） stimulated and induced by high glucose,and to explore the regulatory mechanism of EM-1 on DC immune function.Methods： Peripheral blood mononuclear cells（ PBMNCs） were induced into immature dendritic cells（ imDCs）.The high glucose was used as the stimulating factor,and the EM-1 was used as the interventional factor.Then,the experiments were divided into normal glucose group（ NGgroup）,high glucose group（ HGgroup）,high glucose plus EM-1 group（ EMgroup）and high glucose plus EM-1 and naloxone group（ Nal group）,respectively.The PBDC＇s phenotype changes were detected by flow cytometry; ELISA was used to detect the changes of cytokines secreted by PBDCs co-cultured with autologous lymphocytes; CFSE was used to detect the proliferation of T lymphocytes.TLR4 expression on PBDC surface was detected by RT-PCR.Results： Compared with HGgroup,the expression of PBDC surface molecules CD86,CCR7 and CD36 was up-regulated in EMgroup（ P〈0.01）,while the change of CD83 expression was not statistically significant.However,IL-12 and IL-10 secreted by PBDCs and the proliferation index of T-lymphocytes stimulated by PBDCs were both decreased in EMgroup.Compared with EMgroup,the expression of CD86,CCR7 and CD36 was decreased in Nal group（ P〈0.01）,while the expression of CD83 was almost unchanged（ P〉0.05）.T-lymphocyte proliferation index was increased very significantly in Nal group（ P〈0.01）.The gray ratio of TLR4 in HGgroup was higher than that in NGgroup,while the gray ratio in EMgroup＇s was very significantly lower than that in HGgroup＇s（ P〈0.01）.These results indicate that the high glucose can promote the expression of PBDC TLR4,while the EM-1 inhibits the expression of TLR4.Conclusion： EM-1 up-regulates the expression of PBDC surface molecules CD86,CCR7 and CD36 stimulated and induced by high glucose,but inhibites the induction of PBDC to maturity by high glucose.And the secreted inflammatory cytokines IL-12 and IL-10 inhibites the proliferation of T lymphocytes derived from PBDCs,while naloxone inhibites the effect of EM-1.EM-1 inhibites the expression of TLR4 on PBDC surface induced by high glucose.
作者
刘传苗
杨天华
黄敏
周诚
李永海
李正红
LIU Chuan-Miao;YANG Tian-Hua;HUANG Min;ZHOU Cheng;LI Yong-Hai;LI Zheng-Hong(Department of Infecction, The First Affiliated Hospital of Bengbu Medical College, Bengbu 233004, Anhui Province, China;Laboratory of Physiology, Bengbu Medical College, Bengbu 233030, Anhui Province, China)
出处
《中国实验血液学杂志》
CAS
CSCD
北大核心
2018年第3期886-893,共8页
Journal of Experimental Hematology
基金
supported by Natural Science Foundation of Anhui Province,No.1508085MH170
National Natural Science Foundation of China,No.81472656
Natural Science Foundation Project of Anhui Province Education Department,No.KJ2017A247
