miR-182抑制HES激活Notch信号协调TGF-β信号途径促进急性川崎病

摘要

Objective To explore the relationship between mir-182 inhibiting HES1 activation of Notch signal and TGF- morphological signaling pathway, promoting the occurrence of acute kawasaki disease and its clinical features. Methods the clinical study data of each group of KD were measured by blood routine and the correlation between the expression level of serum mir-182 and the clinical characteristics of children with KD was detected. The proportion of Treg cells in peripheral blood of each group was measured by flow cytometry. The expression levels of Foxp3, TGF-1, TGF-1, TGF-3 RII, Smad3 and Smad4 mRNA in peripheral blood of each group were detected by qrt-pcr. The expression levels of Notchl, Notch3 mRNA, Notch receptor Jagged1 and Jagged2 mRNA in all groups were detected by qrt-pcr. The expression level of HES1 mRNA of the Notch signal pathway was detected by rt-pcr. The target genes of HES1 as mir-182 were detected by using target gene database, luciferase reporter gene method, qrt-pcr and Western blot. The expression of phosphorylated proteins of Notch and TGF- morphological pathway receptors Notch1, Notch3, TGF- morphri, TGF- prii, Smad3 and Smad4 in HES1 treatment group was detected by Western blot. Results the expression of mir-182 was relatively low in the serum of acute group of KD. Mir-182 expression was associated with platelet count in children with acute KD (P ＜ 0. 05). The expression levels of Foxp3, TGF- response, TGF- response RI, TGF- response RII, Smad3, Smad4, Notch1, Notch3, Jagged1 and HES1 mRNA in the KD acute group were significantly lower than those in the KD cure group and the normal group. Mir-182 inhibits mRNA translation and protein expression of HES1. HES1 activates the Notch and TGF-Chinese signaling pathways and induces increased receptor expression. Conclusion mir-182 can affect the Treg cell decrease of the Notch and tgf-ionic signaling pathway by down-regulating the expression of HES1, leading to the promotion of the pathological process of acute kawasaki disease. The targeted regulation of mir-182 and HES1 may become a new target for the detection and treatment of acute kawasaki disease.%目的 探讨miR-182抑制HES1激活Notch信号与TGF-β信号途径,促进急性川崎病的发生及与急性川崎病(KD)临床特征的关系.方法 通过血液常规检测KD各组的临床研究数据并检测血清miR-182表达水平与KD患儿临床特征的相关性;采用流式细胞法检测各组外周血中Treg细胞比例;qRT-PCR检测各组外周血中Foxp3、TGF-β、TGF-βRI、TGF-βRII及Smad3、Smad4 mRNA的表达水平;qRT-PCR检测各组中Notch信号途径配体Notchl、Notch3 mRNA及Notch信号途径受体Jagged1、Jagged2 mRNA的表达水平;RT-PCR检测Notch信号途径靶基因HES1 mRNA的表达水平;利用靶基因数据库、荧光素酶报告基因法、qRT-PCR及Western blot检测HES1为miR-182的靶基因及相关靶向调控关系;Westernblot检测在HES1处理组中Notch及TGF-β途径受体Notch1、Notch3、TGF-βRI、TGF-βRII、Smad3、Smad4的磷酸化蛋白表达.结果 KD急性组血清中miR-182表达量相对较低;miR-182表达与KD急性期患儿的血小板计数有关(P＜0.05);KD急性组Foxp3、TGF-β、TGF-βRI、TGF-βRII、Smad3、Smad4、Notch1、Notch3、Jagged1、HES1 mRNA的表达水平明显低于KD治愈组和正常对照组;miR-182能够抑制HES1的mRNA翻译及其蛋白表达;HES1激活了Notch及TGF-β信号通路,诱导了途径受体表达升高.结论 miR-182可以通过下调HES1的表达影响Notch及TGF-β信号途径Treg细胞减少,导致促进急性川崎病的病理进程,miR-182与HES1的靶向调控可能会成为急性川崎病检测与治疗医学的新靶点.