In Vivo Inhibition of MicroRNA-326 in a NOD.H-2(h4) Mouse Model of Autoimmune Thyroiditis.
In Vivo Inhibition of MicroRNA-326 in a NOD.H-2(h4) Mouse Model of Autoimmune Thyroiditis.
复制标题
自身免疫性甲状腺炎 NOD.H-2(h4) 小鼠模型中 MicroRNA-326 的体内抑制
DOI：
10.3389/fimmu.2021.620916
复制
发表时间：
2021
影响因子：
7.3
通讯作者：
Teng W
中科院分区：
文献类型：
--
作者：
Zhao N;Wang Z;Cui X;Wang S;Fan C;Li Y;Shan Z;Teng W
已有研究报道多种miRNAs参与自身免疫性疾病，但miRNAs在自身免疫性甲状腺炎(AIT)中的潜在调控机制有待进一步探讨。本研究旨在进一步验证miR-326在NOD.H-2H4小鼠中通过尾静脉和甲状腺激素注射慢病毒基因通过ETS-1调节Th17/Treg平衡，从而在AIT中发挥作用。5周龄NOD.H-2H4小鼠随机分为尾静脉注射组和甲状腺注射组，每组分别接受MMU-miR-326海绵(LV-海绵)或慢病毒载体对照。随机分为4组：治疗性LV-ctrl组、治疗性LV-海绵组、预防性LV-ctrl组和预防性LV-海绵组。对照组给予不经静脉注射的高碘水。采用甲状腺苏木精-伊红(HE)染色和酶联免疫吸附试验(ELISA法)检测甲状腺淋巴细胞浸润情况和血清TgAb值。用RT-PCR、Western blotting和流式细胞仪检测Ets-1和淋巴细胞计数。免疫荧光法检测甲状腺CD4+IL-17a+细胞和CD4+ETS-1+细胞，ELISA法检测血清细胞因子。尾静脉注射LV-海绵组小鼠甲状腺炎症评分和血清TgAb滴度显著低于对照组和LV-ctrl组，而脾组织Ets-1蛋白表达明显高于对照组和LV-ctrl组。流式细胞仪检测发现，LV海绵组Th1 7/Treg比值下降，预防性LV海绵组Th1 7/Treg比值显著降低(P=0.036)。免疫荧光显示，LV-海绵组CD_4~+IL-17a~+细胞显著减少(P=0.001)，CD_4~+Ets-1~+细胞显著增加(P=0.029)。LV海绵组血清IL-17/IL-10显著降低(P<0.05)。在甲状腺注射组中，LV-海绵组与LV-ctrl组相比，甲状腺炎症评分和血清TgAb滴度均显著降低(P<；0.05)。免疫荧光检测显示，LV-海绵组CD4+IL-17a+细胞明显减少，而抑制组CD4+Ets-1+细胞显著增加。此外，尾静脉注射LV-海绵后，与甲状腺注射相比，甲炎患者的TgAb水平明显降低。MIR-326靶向治疗AIT可能是一种有前途的治疗方法。另外，尾静脉注射可能比甲状腺注射取得更好的干预效果。
Previous studies reported that various miRNAs participate in autoimmune diseases, but the potential regulatory mechanism of miRNAs in autoimmune thyroiditis (AIT) needs further exploration. This study aimed to further verify that miR-326 contributes to AIT by regulating Th17/Treg balance through Ets-1 using lentiviral gene delivery through tail vein and thyroid injection in NOD.H-2h4 mice. Five-week-old NOD.H-2h4 mice were divided randomly into tail vein and thyroid injection groups, and each received either mmu-miR-326 sponge (LV-sponge) or lentiviral vector control. Mice were divided for tail vein injection: the therapeutic LV-ctrl, therapeutic LV-sponge, prophylactic LV-ctrl, and prophylactic LV-sponge groups. The control group was fed high-iodine water without vein injection. The thyroid infiltration of lymphocytes and serum TgAb value were investigated by thyroid hematoxylin and eosin (HE) staining and ELISA, respectively. Ets-1 and lymphocyte counts were measured by RT-PCR, western blotting, and flow cytometry. The thyroid CD4+IL-17a+ cells and CD4+Ets-1+ cells were detected by immunofluorescence, and the serum cytokines were tested by ELISA. In the tail vein injection groups, the thyroid inflammatory score and serum TgAb titer were significantly lower in the LV-sponge groups than in the control and LV-ctrl groups while Ets-1 protein expression in mouse spleens was increased in the LV-sponge groups. Moreover, Th17/Treg ratio declined in the LV-sponge group and decreased significantly in the prophylactic LV-sponge group (P = 0.036) tested by flow cytometry. Immunofluorescence showed that, in LV-sponge groups, CD4+IL-17a+ cells were decreased significantly (P = 0.001), while CD4+Ets-1+ cells were increased significantly in the LV-sponge group (P = 0.029). The serum IL-17/IL-10 was decreased significantly in the LV-sponge group (P < 0.05). In the thyroid injection groups, the thyroid inflammatory score and serum TgAb titer in the LV-sponge group decreased significantly compared with those in the LV-ctrl group (P < 0.05). In addition, in LV-sponge groups, CD4+IL-17a+ cells were decreased, while CD4+Ets-1+ cells were increased significantly in the inhibition group evaluated by immunofluorescence. Moreover, tail vein injection of LV-sponge resulted in much lower TgAb levels in thyroiditis compared with thyroid injection. MiR-326 targeted therapy may be a promising approach for AIT. In addition, tail vein injection may achieve a better intervention effect than thyroid injection.
登录
查看更多内容
影响因子：
30.5
作者：
Du, Changsheng;Liu, Chang;Pei, Gang
通讯作者：
Pei, Gang
DOI：
10.1084/jem.178.3.793
发表时间：
1993-09-01
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Podolin PL;Pressey A;DeLarato NH;Fischer PA;Peterson LB;Wicker LS
通讯作者：
Wicker LS
自身免疫性甲状腺疾病不同亚型中Th17/Treg失衡
影响因子：
--
作者：
Li, Cui;Yuan, Jianghong;Zhang, Jin-an
通讯作者：
Zhang, Jin-an
microRNA-326 水平升高通过靶向解整合素和金属蛋白酶来调节桥本甲状腺炎中的 IL-23/IL-23R/Th17 细胞轴 17
影响因子：
6.6
作者：
Liu, Yongping;Cui, Xuejiao;Teng, Weiping
通讯作者：
Teng, Weiping
影响因子：
48
作者：
Ebert, Margaret S.;Neilson, Joel R.;Sharp, Phillip A.
通讯作者：
Sharp, Phillip A.