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| 建立II型胶原诱导的Th17细胞体外增殖模型：探索IL-23和胶原活性对自身免疫调节的影响 |
| Establishment of a Collagen Type II-Induced Th17 Cell Proliferation Model In Vitro: Exploring the Effects of IL-23 and Collagen Activity on Autoimmune Regulation |
| 投稿时间：2024-12-30 修订日期：2025-01-13 |
| DOI： |
| 中文关键词: II型胶原 Th17细胞 增殖模型 IL-23 |
| 英文关键词: collagen typeⅡ Th17 cell proliferation model IL-23 |
| 基金项目:三叶青藤总黄酮干预Th17细胞增殖对小鼠胶原性关节炎作用及机制研究,国家自然科学基金项目（面上项目，重点项目，重大项目） |
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| 中文摘要: |
| 【摘要】 目的 建立II型胶原（Collagen Type II, CⅡ）诱导的Th17细胞体外增殖模型，并深入研究不同实验条件对细胞增殖的影响。方法 根据研究要求，收集适量的正常和CIA小鼠脾淋巴细胞，小鼠脾淋巴细胞被随机分为六组，分别接受以下处理：未处理（对照组）、灭活鸡CⅡ刺激、非灭活鸡CⅡ刺激、低（2 ng/ml）、中（10 ng/ml）、高（50 ng/ml）浓度IL-23处理。所有细胞在标准条件下培养60小时后，采用流式细胞术分析CD4+RORt+ Th17细胞的比例。结果体外培养60h后，正常小鼠脾淋巴细胞经灭活或未灭活CⅡ刺激Th17细胞比率均较培养前显著下降，CIA小鼠脾淋巴细胞未用CⅡ刺激Th17细胞比率也较培养前显著下降，而CIA小鼠脾淋巴细胞以灭活CⅡ或未灭活CⅡ刺激后Th17细胞比率均较之前不同程度的提升；该组与对照组横向对比得出差异有统计学意义(p＜0.05)。进一步的分析发现，这一比率在灭活CⅡ与灭活CⅡ的两组之间差异较小(p=0.44)。另外，经过本次研究得出改变IL-23的浓度并未影响体外CIA小鼠Th17比率，然而添加IL-23p19抗体中和试剂的情况下，体外CIA小鼠脾淋巴细胞Th17细胞比率减小，与对照组相比差异显著（P<0.01）。结论 经过展开一系列实验研究，发现灭活与未灭活CⅡ均未能促使体外正常小鼠Th17细胞分裂，然而该方法对体外CIA小鼠Th17细胞增殖有促进作用；IL-23对Th17细胞的体外增殖是必须的。本研究证实此模型确为CⅡ反应性Th17细胞体外增殖模型，该模型可用于抗风湿药物的疗效评价。 |
| 英文摘要: |
| 【Abstract】 Objective To establish a model of reactive Th17 cells proliferation induced by collagen type Ⅱ (C Ⅱ) in vitro and investigate its influencing factors. Methods The splenic lymphocytes of normal and CIA mice were isolated and divided into groups. They were given inactivated or non-inactivated chicken CⅡ or different concentrations of IL-23 (2, 10, 50 ng/ml), or IL-23p19 antibody. Culturing for 60 hours, the ratio of CD4+RORt+Th17 cells was detected by flow cytometry. Then, the results obtained are analyzed, and the corresponding conclusions are drawn.Results After 60 hours of culture in vitro, the ratio of Th17 cells stimulated by inactivated or non-inactivated CⅡ in normal mouse spleen lymphocytes was significantly lower than that before culture, and the ratio of Th17 cells not stimulated by C Ⅱ in CIA mouse spleen lymphocytes was also significantly lower than that before culture, while the ratio of Th17 cells stimulated by inactivated C Ⅱ or non-inactivated C Ⅱ in CIA mouse spleen lymphocytes was significantly higher than that before culture, and there was a significant difference compared with the CIA control group (p<0.05), However, there was no statistical difference in the ratio of Th17 cells between the two groups without inactivated C Ⅱ and inactivated C Ⅱ (p=0.44). After the analysis of the data obtained from the study, it was further concluded that different concentrations of IL-23 did not affect the Th17 cell ratio of spleen lymphocytes of CIA mice in vitro, but after adding IL-23p19 antibody neutralization reagent, the Th17 cell ratio of spleen lymphocytes of CIA mice in vitro decreased significantly, with a statistical?difference compared with the blank control group (P<0.01). Conclusions ?After a series of experimental studies, it was found that both inactivated and non-inactivated C failed to divide Th 17 cells in normal mice. On the contrary, the above method could accelerate the division of Th 17 cells in vitro to varying degrees. This study confirmed that this model is indeed a C Ⅱ reactive Th17 cell proliferation model in vitro, which can be used to evaluate the efficacy of anti-rheumatic drugs，make more patients benefit from it, so it has a certain practical significance. |
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