论文部分内容阅读
目的探讨重组人γ干扰素（ｒｈＩＦＮγ）对人胚胎胰岛分泌白细胞介素６（ＩＬ６）以及对胰岛分泌功能的影响，为ＩＦＮγ和ＩＬ６在１型糖尿病中的可能作用提供依据。方法采用胶原酶Ｖ型消化方法分离人胚胎胰岛，并对其进行原代培养，加入不同浓度的ｒｈＩＦＮγ培养４８小时收获上清，进行ＩＬ６、胰岛素、胰升血糖素活性检测和ＩＬ６单抗中和试验，对ｒｈＩＦＮγ刺激和对照组胰岛ＲＮＡ进行ＩＬ６ＲＮＡ斑点杂交。结果（１）ｒｈＩＦＮγ刺激后胰岛培养上清ＩＬ６含量显著升高，当ｒｈＩＦＮγ浓度在１００ｕ／ｍｌ时ＩＬ６含量最高；（２）ＩＬ６单抗对ｒｈＩＦＮγ刺激的胰岛培养上清中ＩＬ６有很强的中和效应；（３）斑点杂交结果显示ｒｈＩＦＮγ刺激的胰岛ＲＮＡ中ＩＬ６ｍＲＮＡ含量高于对照组；（４）人胚胎胰岛受ｒｈＩＦＮγ刺激后培养上清胰岛素降低不明显；（５）人胚胎胰岛受ｒｈＩＦＮγ刺激后培养上清胰升血糖素分泌能力明显升高，分泌趋势与ＩＬ６相似。结论ｒｈＩＦＮγ可以促进胰岛分泌ＩＬ６，而ＩＬ６的过度分泌可以加重１型糖尿病的进程。
Objective To investigate the effect of recombinant human interferon-gamma (IFN-γ) on the secretion of interleukin 6 (IL6) in human embryonic islets and the function of pancreatic islet secretion, and provide basis for the possible role of IFNγ and IL6 in type 1 diabetes mellitus. Methods Isolation of human embryonic islets by collagenase V digestion method, and primary culture of the islets were carried out. The supernatants were harvested by adding different concentrations of rhIFNγ for 48 hours to detect IL6, insulin, glucagon activity and IL6 Monoclonal antibody neutralization test, rhIFNγ stimulation and control group islet RNA IL 6RNA dot blot. Results (1) The content of IL6 in islet culture supernatant was significantly increased after rhIFNγ stimulation, and IL6 content was the highest when rhIFNγ concentration was 100u / ml; (2) IL6 monoclonal antibody was sensitive to rhIFNγ-stimulated islet culture supernatant IL-6 has a strong neutralizing effect; (3) blot hybridization results show that rhIFNγ-stimulated islet RNA IL 6 mRNA content was higher than that of the control group; (4) human embryonic islet rhIFNγ stimulation cultured supernatant insulin decreased significantly ; (5) Human embryonic islets were stimulated by rhIFNγ after the culture supernatant glucagon secretion was significantly increased secretion trend similar to IL 6. Conclusion rhIFNγ can promote the secretion of IL 6 islets, and IL 6 over-secretion can aggravate the process of type 1 diabetes.