Effect of interrupted endogenous BMP/Smad signaling on growth and steroidogenesis of porcine granulosa cells     

Wei Wang  Li Wang  Xin-xiu Li  Xia Chen  Hai-yan Zhang  Yu He  Jing-jing Wang  Yong-yan Zhao  Bao-le Zhang  Yin-xue Xu 《Journal of Zhejiang University. Science. B》2010,11(9):719-727

Bone morphogenetic proteins (BMPs) play a critical role in the growth and steroidogenesis of granulosa cells (GCs). BMP signals act through membrane-bound heteromeric serine/threonine kinase receptors. Upon ligand binding, BMPs activate intracellular Smad proteins and regulate growth and apoptosis in various cell types. The objective of this study was to demonstrate the effects of BMP/Smad signal on growth and steroidogenesis of porcine GCs. A strategy of RNA interference (RNAi)-mediated ‘gene silencing’ of Smad4, a core molecule mediating the intracellular BMP/Smad signal transduction pathways, was used to interrupt endogenous BMP/Smad signaling. Results indicate that Smad4-small interfering RNA (siRNA) caused specific inhibition of Smad4 mRNA and protein expression after transfection. Interrupted endogenous BMP/Smad signaling significantly inhibited growth, and induced apoptosis of porcine GCs, while decreasing estradiol production. In addition, interrupted BMP/Smad signaling significantly (P<0.05) changed the expression of Cyclin D2, CDK4, Bcl-2, and Cyp19a1. These findings provide new insights into how BMP/Smad signaling regulates the growth and steroidogenesis of porcine GCs.  相似文献   

Effect of high glucose levels on the calcification of vascular smooth muscle cells by inducing osteoblastic differentiation and intracellular calcium deposition via BMP-2/Cbfα-1 pathway     

Liu F  Zhong H  Liang JY  Fu P  Luo ZJ  Zhou L  Gou R  Huang J 《Journal of Zhejiang University. Science. B》2010,11(12):905-911

In this paper,we investigate the effect and the possible mechanism of high glucose levels on the calcification of human aortic smooth muscle cells (HASMCs).HASMCs were divided into four groups: normal glucose group (NG),osmolality control group (OC),high glucose group (HG,HASMCs culture medium containing 30 mmol/L glucose),and high glucose plus recombinant human Noggin protein (bone morphogenetic protein-2 (BMP-2) antagonist) group (HN).The mRNA levels and the protein expressions of BMP-2 and core binding factor alpha-1 (Cbfα-1) were measured by real-time quantitative polymerase chain reaction (PCR) and Western blot.After induced by 10 mmol/L β-glycerol phosphoric acid,cells were harvested for assessments of alkaline phosphatase (ALP) activities at Days 1,2,and 3,and intracellular calcium contents at Days 7 and 14,respectively.High glucose levels increased the mRNA levels and the protein expressions of BMP-2 and Cbfα-1 (P0.05).The expression of Cbfα-1 was partially blocked by Noggin protein (P0.05),while BMP-2 was not (P0.05).After being induced by β-glycerol phosphoric acid,high glucose levels increased the ALP activity [(48.63±1.03) vs.(41.42±2.28) U/mg protein,Day 3;P0.05] and the intracellular calcium content [(2.76±0.09) vs.(1.75±0.07) μmol/mg protein,Day 14;P0.05] in a time-dependent manner when compared with the NG group,while the ALP activity could not be blocked by Noggin protein [(48.63±1.03) vs.(47.37±0.97) U/mg protein,Day 3;P0.05].These results show that high glucose levels can evoke the calcification of HASMCs by inducing osteoblastic trans-differentiation and intracellular calcium deposition via the BMP-2/Cbfα-1 pathway,which can be partially blocked by Noggin protein.  相似文献   

Science Letters：IGFBP7 plays a potential tumor suppressor role against colorectal carcinogenesis with its expression associated with DNA hypomethylation of exon 1   总被引：1，自引：0，他引：1  

Ruan WJ  Lin J  Xu EP  Xu FY  Ma Y  Deng H  Huang Q  Lv BJ  Hu H  Cui J  Di MJ  Dong JK  Lai MD 《Journal of Zhejiang University. Science. B》2006,7(11):929-932

Insulin-like growth factor binding-protein-7 （IGFBP7） was obtained from our previous colonic adenocarcinoma （CRC） and normal mucosa suppression subtraction hybridization （SSH） cDNA libraries. By RT-PCR and immunohistochemistry, we found that IGFBP7 was overexpressed in CRC tissue compared to normal tissue. However, our in vitro experiments performed in 10 CRC cell lines showed that IGFBP7 expressed only in SW480 and Caco2 cell lines, which implied an underlying reversible regulatory mechanism. Using methylation-specific PCR （MSP） and bisulfite sodium PCR （BSP）, we found that its expression was associated with DNA hypomethylation of exonl. This was further supported by the in vitro study which showed restored IGFBP7 expression after demethylation agent 5-aza-2＇-deoxycytidine treatment. Correlation analysis between IGFBP7 expression and prognosis indicated that overexpression of IGFBP7 in CRC tissue correlated with favourable survival. Investigation of the functional role of IGFBP7 through transfection studies showed that IGFBP7 protein could inhibit growth rate, decrease colony formation activity, and induce apoptosis in RKO and SW620 cells, suggesting it a potential tumor suppressor protein in colorectal carcinogenesis. In conclusion, our study clearly demonstrated that IGFBP7 plays a potential tumor suppressor role against colorectal carcinogenesis and its expression is associated with DNA hypomethylation of exon 1.  相似文献   

不同氮肥水平下烯效唑对小麦籽粒氮素含量的影响     

邵庆勤  杨文钰  童春雨  樊高琼 《安徽科技学院学报》2006,20(3):12-16

试验采用两因素裂区设计研究不同氮肥水平下烯效唑对小麦籽粒氮素含量的影响,氮肥总用量为主处理,烯效唑干拌种浓度为副处理,主副处理各设四个水平。结果表明:烯效唑处理后,籽粒中蛋白氮与非蛋白氮含量及最终的蛋白质含量表现一致。在不施氮与低氮水平下,表现为40mg/kg>60mg/kg>20mg/kg>0mg/kg;在中氮与高氮水平下,表现为20mg/kg>0mg/kg>40mg/kg>60mg/kg。相关分析表明,非蛋白氮含量急剧下降阶段即蛋白氮含量急剧上升阶段的蛋白氮与非蛋白氮含量呈极显著负相关;花后10d开始籽粒蛋白氮与收获后籽粒蛋白质含量也显著相关性。因此,烯效唑处理提高了籽粒中的非蛋白氮含量,从而有利于籽粒蛋白氮的合成,进而影响籽粒最终的蛋白质含量。  相似文献   

酶法提取罗非鱼鱼鳍胶原蛋白的研究     

王春  周天  方素云 《茂名学院学报》2013,(6):24-26

文章对罗非鱼鱼鳍胶原蛋白的提取进行研究。通过单因素试验研究鱼鳍胶原蛋白提取的不同因素影响作用,并通过正交试验确定胃蛋白酶酶法提取胶原蛋白的最佳工艺。研究结果表明,在酶解时间6h、底物浓度5％、酶量5500U／g、pH为1．80和温度为40℃的条件下,胃蛋白酶水解罗非鱼鱼鳍效果较好。正交试验表明,酶法提取反应主要因素影响的顺序为：反应时间〉酶量〉底物浓度。  相似文献   

不同品种香菇菌棒蛋白质与多糖含量的比较研究     

徐玉辉  陈海燕 《丽水学院学报》2013,(2):19-21

碱浸提法提取S604、S606、931、中7、808、808对照、868、135、908、9015、939等11种香菇菌棒中的蛋白质和多糖,结果表明：135和908的蛋白质含量显著高于其他9个品种,9015的多糖含量显著高于其他10个品种。  相似文献   

不同抗性的向日葵品种接种锈菌后叶片中可溶性蛋白、可溶性总糖及叶绿素含量的变化   总被引：2，自引：0，他引：2  

景岚  王丽芳  康俊 《临沂师范学院学报》2008,30(6):76-80

通过对两个不同抗性向日葵品种在苗期接种向日葵锈菌,测定了接种后不同时间叶片的可溶性总糖、可溶性蛋白及叶绿素含量的变化．研究结果表明：抗病品种的可溶性总糖比感病品种高;可溶性蛋白质含量、叶绿素b含量和类胡萝卜素含量的变化与向日葵抗病性有密切关系,而和叶绿素a含量的变化与抗病性没有直接关系．  相似文献   

蛋白质空间结构的统计分析     

王昕  毛炳蔚  王福伟  张婧 《雁北师范学院学报》2008,24(5)

利用数据挖掘中的数据分布拟合理论对生物科学领域中的蛋白质侧链空间结构进行统计分析.以世界上广泛使用的生物分子三维结构数据库PDB为基础,利用沈世镒教授提出的对多氨酸残基侧链碳原子间距离的统计分析方法,通过正交试验设计和信息论中的熵函数等相关知识,给出了不同位置、不同氨基酸残基种类对侧链结构的影响.  相似文献   

CHOP及GRP78在复发性多形性胶质母细胞瘤患者表达显著增加     

文雪 《孝感职业技术学院学报》2021,(1):108-112

目的:研究CHOP及GRP78与复发性GBM的相关性。方法:采用Western blot检测CHOP及GRP78在初发及复发GBM组织中的表达;Real-time PCR检测GRP78在初发以及复发GBM组织中的表达。结果:CHOP及GRP78在初发GBM中表达上调,而在经过原发性肿瘤切除后接受放疗和替莫唑胺化疗的复发性GBM中增加尤为显著。结论:CHOP及GRP78在GBM发病机制中起重要作用,该分子有望成为GMB潜在的诊断及预后评估标志物。  相似文献   

Mass spectrometry-based protein-protein interaction techniques and their applications in studies of DNA damage repair     

Zhen CHEN  Junjie CHEN 《Journal of Zhejiang University. Science. B》2021,22(1):1-20

Proteins are major functional units that are tightly connected to form complex and dynamic networks.These networks enable cells and organisms to operate properly and respond efficiently to environmental cues.Over the past decades,many biochemical methods have been developed to search for protein-binding partners in order to understand how protein networks are constructed and connected.At the same time,rapid development in proteomics and mass spectrometry(MS)techniques makes it possible to identify interacting proteins and build comprehensive protein-protein interaction networks.The resulting interactomes and networks have proven informative in the investigation of biological functions,such as in the field of DNA damage repair.In recent years,a number of proteins involved in DNA damage response and DNA repair pathways have been uncovered with MS-based protein-protein interaction studies.As the technologies for enriching associated proteins and MS become more sophisticated,the studies of protein-protein interactions are entering a new era.In this review,we summarize the strategies and recent developments for exploring protein-protein interaction.In addition,we discuss the application of these tools in the investigation of protein-protein interaction networks involved in DNA damage response and DNA repair.  相似文献