中期阿尔茨海默病小鼠模型海马组织Mst1r基因甲基化初探

目的探讨Mst1r基因在中期病程阿尔茨海默病模型(presenilins条件性双基因敲除小鼠,d KO mice)中海马组织的甲基化状态。方法选取12月龄雌性d KO mice及同系野生型小鼠各3只,采用二代测序技术(简化表观亚硫氢酸盐测序技术,RRBS)检测其海马组织基因组DNA以获得甲基化异常基因。结果RRBS测序成果显示中期AD d KO mice海马组织中Mst1r基因呈超甲基化状态(P<0.05)。结论超甲基化的Mst1r基因可能在中期AD d KO mice的病理过程中发挥着一定作用。

Ron receptor-dependent gene regulation of Kupffer cells during endotoxemia

BACKGROUND: Ron receptor tyrosine kinase signaling in macrophages, including Kupffer cells and alveolar macrophages,suppresses endotoxin-induced proinflammatory cytokine/chemokine production. Further, we have also identified genes from Ron replete and Ron deplete livers that were differentially expressed during the progression of liver inflammation associated with acute liver failure in mice by microarray analyses.While important genes and signaling pathways have been identified downstream of Ron signaling during progression of inflammation by this approach, the precise role that Ron receptor plays in regulating the transcriptional landscape in macrophages, and particular in isolated Kupffer cells, has still not been investigated.METHODS: Kupffer cells were isolated from wild-type(TK+/+)and Ron tyrosine kinase deficient(TK-/-) mice. Ex vivo, the cells were treated with lipopolysaccharide(LPS) in the presence or absence of the Ron ligand, hepatocyte growth factor-like protein(HGFL). Microarray and qRT-PCR analyses were utilized to identify alterations in gene expression between genotypes.RESULTS: Microarray analyses identified genes expressed differentially in TK+/+ and TK-/- Kupffer cells basally as well as after HGFL and LPS treatment. Interestingly, our studies identified Mefv, a gene that codes for the anti-inflammatory protein pyrin, as an HGFL-stimulated Ron-dependent gene.Moreover, lipocalin 2, a proinflammatory gene, which is induced by LPS, was significantly suppressed by HGFL treatment.Microarray results were validated by qRT-PCR studies on Kupffer cells treated with LPS and HGFL.CONCLUSION: The studies herein suggest a novel mechanism whereby HGFL-induced Ron receptor activation promotes the expression of anti-inflammatory genes while inhibiting genes involved in inflammation with a net effect of diminished inflammation in macrophages.

Ron receptor-dependent gene regulation in a mouse model of endotoxin-induced acute liver failure

BACKGROUND:Prior experimentation has shown that loss of the tyrosine kinase(TK) signaling domain of the Ron receptor leads to marked hepatocyte protection in a model of lipopolysaccharideinduced acute liver failure(ALF) in D-galactosamine(GalN)sensitized mice.The aim of this study was to identify the role of Ron in the regulation of hepatic gene expression.METHODS:Microarray analyses were performed on liver RNA isolated sequentially from wild-type(WT) and TK-/mice during the progression of ALF.Gene array data were validated using Western and immunohistochemistry analyses as well as with ex vivo culture systems.RESULTS:At baseline,101 genes were differentially expressed between WT and TK-/-livers,which regulate processes involved in hypoxia,proliferation,apoptosis and metabolism.One hour after ALF induction,WT livers exhibited increased cytokine expression compared to TK-/-livers,and after 4 hours,an induction of suppressor of cytokine signaling(SOCS) genes as well as JAK-STAT pathway activation were prominent in TK-/livers compared to controls.CONCLUSION:Our studies suggest a novel hepato-protective mechanism in Ron TK-/-mice wherein increased and sustained SOCS production and JAK-STAT activation in the hepatocyte may inhibit the destructive proinflammatory milieu and promote survival factors which blunt hepatic death and the ensuing development of ALF.

RON基因在膀胱癌组织中的表达及对T24细胞增殖的影响

目的探讨RON基因对T24细胞增殖与迁移的影响。方法收集膀胱癌组织标本,通过RT-q PCR检测RON在膀胱癌组织中的表达,其次用siRNA技术干扰T24细胞RON,用RT-q PCR、Western blot检测干扰效果,CCK-8、Transwell实验及流式细胞术分别检测细胞的增殖、迁移及细胞凋亡。结果膀胱癌组织中RON表达与TNM分期呈正相关(P<0.05)。在T24细胞的RON-siRNA组,细胞增殖能力减弱(P<0.05),迁移与侵袭能力降低(P<0.05),凋亡细胞数增加(P<0.05),同时P-ERK蛋白表达减少,但Total-ERK表达不变。结论 RON基因促进膀胱癌细胞的增殖与迁移,有望成为膀胱癌治疗的新靶点。