Unconjugated
Emerging evidence has shown that microRNAs have key roles in regulating various normal physiological processes, whereas their deregulated expression is correlated with various diseases. The miR-146 family includes miR-146a and miR-146b, with a distinct expression spectrum in different hematopoietic cells. Recent work indicated that miR-146a has a close relationship with inflammation and autoimmune diseases. miR-146-deficient mice have developed some abnormal hematopoietic phenotypes, suggesting the potential functions of miR-146 in hematopoietic development. In this study, we found that miR-146b was consistently up-regulated in both K562 and CD34(+) hematopoietic stem/progenitor cells (HSPCs) undergoing either erythroid or megakaryocytic differentiation. Remarkably, erythroid and megakaryocytic maturation of K562 cells was induced by excess miR-146b but inhibited by decreased miR-146b levels. More importantly, an mRNA encoding receptor tyrosine kinase, namely platelet-derived growth factor receptor α (PDGFRA), was identified and validated as a direct target of miR-146b in hematopoietic cells. Gain-of-function and loss-of-function assays showed that PDGFRA functioned as a negative regulator in erythroid and megakaryocytic differentiation. miR-146b could ultimately affect the expression of the GATA-1 gene, which is regulated by HEY1 (Hairy/enhancer-of-split related with YRPW motif protein 1), a transcriptional repressor, via inhibition of the PDGFRA/JNK/JUN/HEY1 pathway. Lentivirus-mediated gene transfer also demonstrated that the overexpression of miR-146b promoted erythropoiesis and megakaryocytopoiesis of HSPCs via its regulation on the PDGFRA gene and effects on GATA-1 expression. Moreover, we confirmed that the binding of GATA-1 to the miR-146b promoter and induction of miR-146b during hematopoietic maturation were dependent on GATA-1. Therefore, miR-146b, PDGFRA, and GATA-1 formed a regulatory circuit to promote erythroid and megakaryocytic differentiation.
The purpose of this study is to examine germ cells apoptosis and reduction of spermatogenesis which might be induced by deltamethrin (DM). Furthermore, the study is performed to determine if the apoptosis is mediated by the signaling proteins: eNOS, JNK1 and androgen receptor (AR). Fifty-four male SD rats were divided into nine groups (six rats each): blank control group; corn oil treated group; DM treated group; saline treated group; DM+saline treated group; DM+histamine (eNOS specific agonist) treated group; 50% ethanol treated group; DM+50% ethanol group and DM+quercetagetin (JNK1 specific inhibitor) treated group. The experiment was conducted for 15 days. Apoptosis was evaluated by TUNEL; S-nitrosylation of JNK1 was examined by the biotin switch assay; eNOS expression and Ser650 phosphorylation of AR were assessed by immunoblotting and immunohistochemical analysis, respectively. DM treated group showed notable apoptotic cells and reduced production of sperm, while DM plus histamine group and DM plus quercetagetin group showed remarkably decreased apoptosis and improved production of sperm. Administration of DM inhibited spermatogenesis, the activity of eNOS and S-nitrosylation of JNK1. Meanwhile, phosphorylation of AR was shown to be elevated. Histamine and quercetagetin were also examined to have a further confirmation. It is suggested DM-induced germ cells apoptosis and reduction of sperm production were mediated by eNOS-JNK1-AR signaling pathway.