中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2015年
6期
442-447
,共6页
卢龙坤%韦腾飞%朱力%吴豫%沈茜
盧龍坤%韋騰飛%硃力%吳豫%瀋茜
로룡곤%위등비%주력%오예%침천
瘦素%Th17细胞%白细胞介素17%RORγt
瘦素%Th17細胞%白細胞介素17%RORγt
수소%Th17세포%백세포개소17%RORγt
Leptin%Th17 cell%IL-17%RORγt
目的：探讨瘦素对Th17细胞生成的影响及相关机制。方法以瘦素基因敲除的ob／ob小鼠和同系野生小鼠为研究对象。流式细胞术检测Th17细胞比例，免疫磁珠法分选脾脏CD4＋T细胞，在体外培养中拮抗瘦素或添加不同浓度的瘦素以观察瘦素对Th17细胞生成的影响，实时荧光定量逆转录聚合酶链反应（ qPCR）和Luminex检测技术分别检测经体外培养72 h后小鼠脾脏CD4＋T细胞中与Th17细胞密切相关细胞因子等mRNA相对表达水平及培养上清液中蛋白的表达水平。结果 ob／ob鼠外周血、脾脏中Th17细胞的比例均显著低于野生鼠[（0．49±0．03）％ vs （1．29±0．1）％、（1．56±0．22）％ vs （2．47±0．11）％]；体外培养中，野生鼠脾脏CD4＋T细胞经瘦素抗体处理后，Th17细胞的比例明显减少[（2．04±0．11）％vs （3．39±0．31）％]；ob／ob鼠脾脏CD4＋T细胞加入不同浓度瘦素后，Th17细胞比例则以剂量依赖的形式增加[（2．15±0．25）％，（3．12±0．37）％ vs （1．41±0．13）％]；ob／ob鼠脾脏CD4＋T细胞经体外培养后RORγt、IL-17A和IL-6 mRNA的相对表达量及上清中IL-17A和IL-6蛋白表达水平均低于野生鼠，但TGF-β和IL-23 mRNA表达水平没有差异。结论瘦素缺乏可导致小鼠Th17细胞的生成显著减少，也造成RORγt、IL-17A和IL-6 mRNA及IL-17和IL-6蛋白的表达水平显著降低。瘦素可能通过上调RORγt和IL-6的转录，促进Th17细胞的生成及效应分子分泌。
目的：探討瘦素對Th17細胞生成的影響及相關機製。方法以瘦素基因敲除的ob／ob小鼠和同繫野生小鼠為研究對象。流式細胞術檢測Th17細胞比例，免疫磁珠法分選脾髒CD4＋T細胞，在體外培養中拮抗瘦素或添加不同濃度的瘦素以觀察瘦素對Th17細胞生成的影響，實時熒光定量逆轉錄聚閤酶鏈反應（ qPCR）和Luminex檢測技術分彆檢測經體外培養72 h後小鼠脾髒CD4＋T細胞中與Th17細胞密切相關細胞因子等mRNA相對錶達水平及培養上清液中蛋白的錶達水平。結果 ob／ob鼠外週血、脾髒中Th17細胞的比例均顯著低于野生鼠[（0．49±0．03）％ vs （1．29±0．1）％、（1．56±0．22）％ vs （2．47±0．11）％]；體外培養中，野生鼠脾髒CD4＋T細胞經瘦素抗體處理後，Th17細胞的比例明顯減少[（2．04±0．11）％vs （3．39±0．31）％]；ob／ob鼠脾髒CD4＋T細胞加入不同濃度瘦素後，Th17細胞比例則以劑量依賴的形式增加[（2．15±0．25）％，（3．12±0．37）％ vs （1．41±0．13）％]；ob／ob鼠脾髒CD4＋T細胞經體外培養後RORγt、IL-17A和IL-6 mRNA的相對錶達量及上清中IL-17A和IL-6蛋白錶達水平均低于野生鼠，但TGF-β和IL-23 mRNA錶達水平沒有差異。結論瘦素缺乏可導緻小鼠Th17細胞的生成顯著減少，也造成RORγt、IL-17A和IL-6 mRNA及IL-17和IL-6蛋白的錶達水平顯著降低。瘦素可能通過上調RORγt和IL-6的轉錄，促進Th17細胞的生成及效應分子分泌。
목적：탐토수소대Th17세포생성적영향급상관궤제。방법이수소기인고제적ob／ob소서화동계야생소서위연구대상。류식세포술검측Th17세포비례，면역자주법분선비장CD4＋T세포，재체외배양중길항수소혹첨가불동농도적수소이관찰수소대Th17세포생성적영향，실시형광정량역전록취합매련반응（ qPCR）화Luminex검측기술분별검측경체외배양72 h후소서비장CD4＋T세포중여Th17세포밀절상관세포인자등mRNA상대표체수평급배양상청액중단백적표체수평。결과 ob／ob서외주혈、비장중Th17세포적비례균현저저우야생서[（0．49±0．03）％ vs （1．29±0．1）％、（1．56±0．22）％ vs （2．47±0．11）％]；체외배양중，야생서비장CD4＋T세포경수소항체처리후，Th17세포적비례명현감소[（2．04±0．11）％vs （3．39±0．31）％]；ob／ob서비장CD4＋T세포가입불동농도수소후，Th17세포비례칙이제량의뢰적형식증가[（2．15±0．25）％，（3．12±0．37）％ vs （1．41±0．13）％]；ob／ob서비장CD4＋T세포경체외배양후RORγt、IL-17A화IL-6 mRNA적상대표체량급상청중IL-17A화IL-6단백표체수평균저우야생서，단TGF-β화IL-23 mRNA표체수평몰유차이。결론수소결핍가도치소서Th17세포적생성현저감소，야조성RORγt、IL-17A화IL-6 mRNA급IL-17화IL-6단백적표체수평현저강저。수소가능통과상조RORγt화IL-6적전록，촉진Th17세포적생성급효응분자분비。
Objective To investigate the effects of leptin on Th17 cells and the possible mechanism. Methods The leptin-deficient ( ob/ob) mice and their homologous wild-type mice were used in the study.The percentages of Th17 cells in peripheral blood samples, spleen tissues and lymph nodes were measured by flow cytometry ( FCM) analysis.The splenic CD4+T cells, separated from the ob/ob mice and the wild-type mice by using magnetic beads,were respectively cultured with leptin at various concentrations and with anti-leptin neu-tralization antibody to evaluate the effects of leptin on Th17 cells.The quantitative real-time PCR was performed to analyze Th17 cell-related cytokines at transcriptional levels.The levels of IL-6 and IL-17A in the supernatants of CD4+T cell culture were measured with Luminex technology.Results Compared with the wild-type mice, the ob/ob mice showed lower percentages of Th17 cells in both peripheral blood samples and spleen tissues (0.49%±0.03%vs 1.29%±0.1%, 1.56%±0.22%vs 2.47%±0.11%).There was a decrease in the percentages of Th17 cells upon the in vitro treatment of CD4+T cells from wild-type mice with anti-leptin antibody.The per-centages of Th17 cells were increased in a dose-dependent manner upon the in vitro treatment of CD4+T cells from ob/ob mice with leptin.Moreover, the levels of IL-17A and IL-6 and the transcriptional levels of RORγt, IL-17A and IL-6 in leptin deficiency group were lower than those of wild-type group, but were increased upon the treatment with leptin.No significant difference with the transcriptional levels of TGF-βand IL-23 was ob-served between the two groups with and without intervention.Conclusion Leptin deficiency seriously hampered the generation of Th17 cells in mice and resulted in a decreased expression of RORγt, IL-17A and IL-6 at mRNA level.The treatment of CD4 T cells with leptin might promote the generation of Th17 cells through up-regulating the transcription of RORγt and IL-6.
