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AIM To investigate the role of fatty acid translocase (FAT/CD36) on differentiation of monocytes to macrophages. METHODS Human monocyte THP-1 cells were treated with phorbol 12-myristate 13-acetate (PMA) at 0, 100 and 200 μg /L. Small interfering RNA (siRNA) targeting CD36 (siCD36) was employed to knock down the expression of CD36 in THP-1 cells. The CD36 over-expression (CD36OE) cell line was constructed by transfection with a recombinant lentivirus containing CD36 cDNA. Optical microscopy and crystal violet staining were used to detect the monocyte morphological changes and adhesion ability. The protein expression of CD36 was measured by flow cytometry and Western blot. The mRNA levels of CD36, CD11b and CD80 were detected by real-time PCR. The protein levels of extracellular signal-regulated kinase (ERK） and Src tyrosine kinase were determined by Western blot. RESULTS The cellular adhesiveness of THP-1 cells was elevated in the process of monocytes differentiation, and the expression of CD36 was increased in this process as well (P <0.01). siCD36 was transfected into the THP-1 cells (CD36i group) and the silencing efficiency was approximately 80%. The cell surface area and cellular adhesiveness were significantly decreased in CD36i group compared with scrambled siRNA (NCi) group (P <0.01). The mRNA levels of CD11b and CD80 were decreased in CD36i group compared with NCi group (P <0.01). The cell surface area and cellular adhesiveness were increased in CD36OE group compared with empty vector (vector) group (P <0.05). The mRNA levels of CD11b and CD80 were increased in CD36OE group compared with vector group (P <0.01). The phosphorylation levels of ERK and Src were decreased in CD36i group compared with NCi group (P <0.05). CONCLUSION CD36 promotes the differentiation of human monocyte THP-1 cells to macrophages by increasing the phosphorylation of Src and further activating ERK. 相似文献
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采用巢式PCR、DNA测序、创造酶切位点法PCR(Created restriction site-PCR,CRS-PCR)及PCR-RFLP(Poly-merase chain reaction-restriction fragment length polymorphism assay)方法,研究中国荷斯坦牛、鲁西黄牛及渤海黑牛的Src基因内含子6,8,9的单核苷酸多态性(SNPs),发现了14062(C/T)、17302(G/A)和18107(T/C)共3个SNPs,均为首次报道的位点。Src基因14062(C/T)、17302(G/A)和18107(T/C)3个位点在中国荷斯坦牛和鲁西黄牛两个群体中的优势等位基因相同,分别为C、G、C,其等位基因频率分别为0.671/0.647、0.583/0.640、0.558/0.577,而渤海黑牛在这三个位点的优势等位基因分别为T、G、C,其等位基因频率分别为0.525,0.913,0.763。经χ2适合性检验,中国荷斯坦牛、鲁西黄牛和渤海黑牛三个群体在14062(C/T)位点均达到Hardy-Weinberg平衡状态(P0.05);在17302(G/A)位点则均未达到Hardy-Weinberg平衡状态(P0.05);中国荷斯坦牛和鲁西黄牛在18107(T/C)位点均达到Hardy-Weinberg平衡状态(P0.05),而渤海黑牛未达到Hardy-Weinberg平衡状态(P0.05)。中国荷斯坦牛和鲁西黄牛在14062(C/T)和18107(T/C)位点均表现为中度多态(0.25PIC0.5);渤海黑牛在17302(G/A)位点表现为低度多态,而其他2个品种牛表现为中度多态。 相似文献
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生长激素(growth hormone,GH)对动物的生长发育具有重要的生理作用.GH与生长激素受体(growth hormone receptor,GHR)结合后才会发挥一系列的生理作用.近年来,人们对GHR结构和功能的研究取得了巨大的进展,并取得了一些重大的突破.现在已清楚了GH-GHR轴激活一些相关的信号转导通路,但并非所有的通路都依赖酪氨酸激酶.作者从以下几个方面总结了GHR作用下的信号转导机制的研究进展:GHR的结构与功能;依赖JAK2的相关信号通路;不依赖JAK2的相关信号通路;GHR信号负调控因子.阐明这些复杂机制,对进一步了解GH对动物不同的生理和病理作用具有重要意义. 相似文献
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AIM: To study the effects of Src suppressed C kinase substrates(SSeCKS) on the adhesion and migration of bovine pulmonary artery endothelial cells (BPAECs) stimulated by fibronectin(FN).METHODS: Cultured BPAEC were stimulated by different concentrations of FN.SSeCKS expressions were detected by Western blotting.Cultured BPAECs were treated with Ro31-8220 and calphostin C (a PKC inhibitor).The locations of SSeCKS,F-actin and vinculin before and after stimulation were observed by confocal microscopy.RESULTS: After stimulated by FN,SSeCKS expression in BPAECs increased in concentration and time dependent manners.After treated with Ro31-8220 and calphostin C,the adhesion and migration of BPVECs were restrained,the expression of SSeCKS was inhibited,SSeCKS distributed from cytosol to perinuclear,and the colocalization of SSeCKS with F-actin,vinculin at the edge of BPAECs was reduced.CONCLUSION: SSeCKS may play an important role in ECs adhesion and migration stimulated by fibronectin.The process can be inhibited by Ro31-8220 and calphostin C. 相似文献
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Role of protein tyrosine phosphatase SHP-2 in cardiac fibroblasts proli-feration stimulated by Ang Ⅱ
ZHANG Li-jing JIN Cheng-yan WANG Guo-yuan WU Bo LI Quan-feng XU Chang-qing TIAN Ye ZHANG Li 《园艺学报》2012,28(2):244-248
AIM: To investigate the potential role of Src homology 2 domain-containing protein tyrosine phosphatase 2 (SHP-2) in the proliferation of cardiac fibroblasts induced by angiotensin Ⅱ (Ang Ⅱ). METHODS: The neonatal rat cardiac fibroblasts were separated by trypsin digestion. The cardiac fibroblasts were identified by vimentin of immunochemical staining. The proliferation of the cardiac fibroblasts with or without Ang Ⅱ stimulation was measured by MTT assay. Adenoviruses containing SHP-2 gene were transfected into cardiac fibroblasts to overexpress SHP-2. SHP-2 was inhibited by its inhibitor NSC-87877. RESULTS: The proliferation of the cardiac fibroblasts was increased in a dose-dependent manner by the stimulation of Ang Ⅱ and the maximum concentration of Ang II for cell proliferation was 10-7 mol/L. SHP-2 promoted the proliferation of cardiac fibroblasts under the stimulation of Ang Ⅱ. The proliferation rate in mutant group was higher than that in wild-type group (P<0.01). Inhibition of SHP-2 by NSC-87877 attenuated the proliferation. CONCLUSION: The growth promoting effect of Ang Ⅱ on cardiac fibroblasts is regulated by SHP-2. 相似文献
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Tesseraud S Métayer S Duchêne S Bigot K Grizard J Dupont J 《Domestic animal endocrinology》2007,33(2):123-142
Insulin induces protein accretion by stimulating protein synthesis and inhibiting proteolysis. However, the mechanisms of regulation of protein metabolism by insulin are complex and still not completely understood. The use of approaches combining hyperinsulinemic clamp and isotopic methods, or measurement of the activation of intracellular kinases involved in insulin signaling, in addition to the use of different animal models in a comparative physiology process, provide better understanding of the potential regulation of protein metabolism by insulin. Studies using the clamp technique in lactating goats have shown a clear inhibitory effect of insulin on proteolysis, with an interaction between the effects of insulin and amino acids. Such studies revealed that the insulin-inhibited proteolysis is improved in lactating goats, this adaptative process limiting the mobilization of body protein under the conditions of amino acid deficit which occurs during early lactation. Insulin signaling studies in growing chickens have also provided some interesting features of insulin regulation compared to mammals. Refeeding or insulin injection leads to the activation of the early steps of insulin receptor signaling in the liver but not in the muscle. Muscle p70 S6 kinase, a kinase involved in the insulin activation of protein synthesis, was found to be markedly activated in response to insulin and to refeeding, suggesting that other signaling pathways than those classically described in mammalian muscles may be involved in signal transduction. Finally, although the role of insulin has been doubtful and has long been considered to be minor in ruminants and in avian species, this hormone clearly regulates protein metabolism in both species. 相似文献
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在体外构建一个以Src蛋白为靶位点的蛋白酪氨酸激酶抑制剂快速筛选模型,为筛选Src蛋白酪氨酸激酶抑制剂奠定基础.基因工程表达GST-v-Src蛋白,收集包涵体蛋白,并经变性复性处理.以生物素化的聚Glu∶Tyr(4∶1)为激酶反应底物、以辣根过氧化物酶(horseradish peroxidase,HRP)标记的磷酸酪氨酸特异性单克隆抗体(HRP-PY20)检测磷酸酪氨酸残基的酶联免疫吸附实验(enzyme-linked immunosorbent assay,ELISA)测定获得蛋白的激酶活性,并进行抑制剂筛选.该模型具有靶向性强、快速、简便可行、高通量的特点.用该方法对6种治疗肿瘤配方中常用的中草药进行筛选,筛选出3种中草药水煎剂富含Src蛋白酪氨酸激酶抑制剂成分. 相似文献
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Chun-Yen Chen Chin-Yang Chang Hung-Jen Liu Ming-Huei Liao Chi-I Chang Jue-Liang Hsu Wen-Ling Shih 《Veterinary research》2010,41(2)
Our previous report demonstrated that bovine ephemeral fever virus (BEFV)-infected cultured cells could induce caspase-dependent apoptosis. This study aims to further elucidate how BEFV activates the caspase cascade in bovine cells. BEFV replicated and induced apoptosis in Vero and Madin-Darby bovine kidney (MDBK) cells, and a kinetic study showed a higher efficiency of replication and a greater apoptosis induction ability of BEFV in Vero cells. Src and c-Jun N-terminal kinase (JNK) inhibitor, but not extracellular signal-regulated kinase (ERK) or p38 inhibitor, alleviated BEFV-mediated cytopathic effect and apoptosis. In BEFV-infected Vero and MDBK cells, BEFV directly induced Src tyrosine-418 phosphorylation and JNK phosphorylation and kinase activity, which was inhibited specifically by SU6656 and SP600125, respectively. The caspase cascade and its downstream effectors, Poly (ADP-ribose) polymerase (PARP) and DFF45, were also activated simultaneously upon BEFV infection. In addition, cytochrome c, but not Smac/DIABLO, was released gradually from mitochondria after BEFV infection. SU6656 suppressed Src, JNK, and caspase-3 and -9 activation, as well as PARP and DFF45 cleavage; SP600125 reduced JNK and caspase-3 and -9 activation, as well as PARP and DFF45 cleavage. Taken together, these results strongly support the hypothesis that a Src-dependent JNK signaling pathway plays a key role in BEFV-induced apoptosis. The molecular mechanism identified in our study may provide useful information for the treatment of BEFV. 相似文献
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