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ZHAO Shu-Jie, BI Mei, Fan Chen-Ling, LIU Miao, LIU Tong, LI Yu-Shu , SHAN Zhong-Yan, TENG Wei-Ping

The study of microRNAs change in Iodine-induced autoimmune thyroiditis model of NOD.H-2 h4 mice. ZHAO Shu-Jie, BI Mei, Fan Chen-Ling, LIU Miao, LIU Tong, LI Yu-Shu , SHAN Zhong-Yan, TENG Wei-Ping The Endocrine Department of the First Affiliated Hospital of China Medical University,

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ZHAO Shu-Jie, BI Mei, Fan Chen-Ling, LIU Miao, LIU Tong, LI Yu-Shu , SHAN Zhong-Yan, TENG Wei-Ping

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  1. The study of microRNAs change in Iodine-induced autoimmune thyroiditis model of NOD.H-2h4 mice ZHAO Shu-Jie, BI Mei, Fan Chen-Ling, LIU Miao, LIU Tong,LI Yu-Shu, SHAN Zhong-Yan, TENG Wei-Ping The Endocrine Department of the First Affiliated Hospital of China Medical University, the Endocrine Institute of China Medical University, the Liaoning Provincial Key Laboratory of Endocrine Diseases

  2. BACKGROUND Autoimmune thyroiditis (AIT) is one of typical organ specific autoimmune diseases. Patients with AIT are characterized by lymphocytes infiltration in thyroid gland, destruction of thyroid follicular cells, serum autoantibodies such as TPOAb and TgAb in serum, and hypofunction.

  3. BACKGROUND In present opinion, immune cells (especially T cells and B cells) abnormality is immunological basis in AIT. Possible mechanisms contain: Abnormal Th1/Th2 Functional deficiency of Ts 、Treg Abnormal Th17 Cloned proliferation of autoreactive T cells and B cells

  4. BACKGROUND Control 7.98% AIT 3.16% APC-CD25 APC-CD25 PE-Foxp3 PE-Foxp3 Our previous study: CD4+CD25+Foxp3+ Treg cells in spleensfrom iodine-treatedNOD.H-2h4 mice and controls. SSC-H FITC-CD4

  5. BACKGROUND AIT 2.45% Control 0.89% PE-IL-17 PE-IL-17 FITC-CD4 FITC-CD4 IL-17 expression in splenocytes from iodine-treatedNOD.H-2h4 mice and controls.

  6. # # # # 结果 5 BACKGROUND Foxp3, IL-17 mRNA expression in splenocytes from iodine-treatedNOD.H-2h4 mice and controls. #:P<0.05

  7. BACKGROUND MicroRNAs (miRNAs) have recently emerged as a major class of gene expression regulators linked to most biological functions. Several miRNAs have been identified as importantregulators for immune cell development, as well as immuneresponse.

  8. BACKGROUND Recent studies have demonstrated that miR-155 plays a crucial role in the function of pathogenic immune cells, including Th1 cells, Treg cells, B cells, DCs and Th17 cells. O’connell RM. Nat Reviews Immunol 2010,22: 111-122

  9. BACKGROUND It was reported that miR-326 is critical in regulating TH-17 differentiation and hence contributes to the pathogenesis of multiple sclerosis. Knockdown or overexpression of miR-326 alleviated or aggravated EAE, respectively. Martin AJ. Nat Immunol 2009,10: 1229-1231

  10. BACKGROUND MiR-146a is among the miRNAs prevalently expressed in Treg cells and showed that it is critical for Treg functions. ZHOU L. Cell & Molecul Biological 2011,8: 380-381

  11. OBJECTIVES The aim of this study was to observe theexpression of miRNA-155, miRNA-146 and miRNA-326in splenic CD4+ T cells and thyroid tissueof NOD.H-2h4 mice with iodine-induced autoimmune thyroiditis.

  12. MATERIAL AND METHOD iodine-treated groups（n=10） 0.05% NaI solution （12week） get splenic CD4+T cells NOD.H-2h4 female mice at 4 weeks of age sterile water （12week） control group （n=6） randomly divided iodine-treated groups（n=10） 0.05% NaI solution （20week） get thyroid tissue sterile water （20week） control group （n=8）

  13. MATERIAL AND METHOD Mice were sacrificed at the time point of 12 weeks after the beginning of experiment. get thyroid get spleen routinely separate mononuclear cells The severity of lymphocytic infiltration was observed using HE-stained thyroid sections. CD4+T cells were be purified by MACS stained with multicolor immunofluorescence And detected by flowcytometry to analysis the percentage of CD4+CD3e+cells Target micoRNA Expression of splenic CD4+T cells were Measuredby Real-time RT-PCR.

  14. MATERIAL AND METHOD Mice were sacrificed at the time point of 20 weeks after the beginning of experiment. get thyroid get another thyoid The severity of lymphocytic infiltration was observed using HE-stained thyroid sections. homogenete Target micoRNA expression In thyroid were measured by Real-time RT-PCR.

  15. RESULTS Lymphocytic infiltration was observed in NOD.H-2h4 mice fed with high iodine water for 12 weeks and 20 weeks(P < 0.01). Table.1 The rate of thyroiditisof thyroiditis after NOD-2h4 mice were feed with sterile water and iodine water at the time point of 12 and 20 weeks（%） #P < 0.01，1000HI：0.05% NaI solution group(about 1000 times of normal daily iodine intake amount, 1000HI)

  16. RESULTS Fig.1 HE-stained thyroid sections after NOD.H-2h4 mice were fed with sterile water. (100X and 200X)

  17. RESULTS Fig.2 HE-stained thyroid sections after NOD.H-2h4 mice were fed with 0.05%NaI for 12week . (100X and 200X)

  18. RESULTS Fig.3 HE-stained thyroid sections after NOD.H-2h4 mice were fed with 0.05%NaI for 20 weeks . (100X and 200X)

  19. 0.41% 79.95% PE-CD3e FITC-CD4 RESULTS Fig.4 Splenic CD4+ Tcell separation ratio was analyzed by Immunofluorescence stain flow cytometric analysis. CD4+ Tcell (left) and rest of cells(right).

  20. RESULTS Table.2 12 weeksafter the beginning of experiment. miRNA-155, miRNA-146, miRNA-326 in splenic CD4+ Tcells were quantified by realtime quantitative PCR.

  21. RESULTS After 12th week, compared with control group（1.418±0.9665）, mice with AIT（4.974±4.324） have rised expression of miRNA-155 in splenic CD4+T cells (P < 0.05)

  22. RESULTS After 12th week, compared with control group（0.9650±0.9347）, mice with AIT（3.332±3.085） have rised expression of miRNA-146 in splenic CD4+T cells (P < 0.05)

  23. RESULTS After 12th week, compared with control group（1.039±0.3910）, mice with AIT（2.534±2.666），there is no obvious change in expression of miRNA-326in splenic CD4+T cells (P > 0.05)

  24. RESULTS Table.3 20 weeksafter the beginning of experiment. miRNA-155, miRNA-146, miRNA-326 in thyroid tissues were quantified by realtime quantitative PCR.

  25. RESULTS After20th week, compared with control group（0.1781±0.1234）, mice with AIT（0.6929±0.2821） have rised expression of miRNA-155 in thyroid tissues (P < 0.001)

  26. RESULTS After20th week, compared with control group（0.2656±0.2350）, mice with AIT（0.9529±0.2619） have rised expression of miRNA-146 in thyroid tissues (P < 0.001)

  27. RESULTS After20th week, compared with control group（0.5103±0.1898）, mice with AIT（1.585±0.4035） have rised expression of miRNA-326 in thyroid tissues (P < 0.001)

  28. CONCLUSIONS Expression of miRNA-155, miRNA-146 and miRNA-326 were higher in both splenic CD4+ T cells and thyroid tissue of NOD.H-2h4 mice with iodine-induced autoimmune thyroiditis than in control mice. miRNA-155, miRNA-146 and miRNA-326 may play important role in the pathogenesis of AIT. Further study are needed for over-expression or down-regulation of those microRNAs in AIT.

  29. Thank You!

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