Profiling of the secreted proteins during 3T3-L1 adipocyte differentiation leads to the identification of novel adipokines

Adipose tissue is an endocrine organ capable of secreting a number of adipokines with a role in the regulation of adipose tissue and whole-body metabolism. We used two-dimensional gel electrophoresis combined with mass spectrometry to profile the secreted proteins from (pre)adipocytes. The culture medium of 3T3-L1 cells during adipocyte differentiation was screened, and 41 proteins that responded to blocking of secretion by 20°C treatment and/or brefeldin A treatment were identified. Prohibitin, stress-70 protein, and adhesion-regulating molecule 1 are reported for the first time as secreted proteins. In addition, procollagen C-proteinase enhancer protein, galectin-1, cyclophilin A and C, and SF20/IL-25 are newly identified as adipocyte secreted factors. Secretion profiles indicated a dynamic environment including an actively remodeling extracellular matrix and several factors involved in growth regulation.Received 15 June 2004; received after revision 26 July 2004; accepted 2 August 2004     

枯草芽孢杆菌谷氨酰胺转氨酶的异源表达

以野生型枯草芽孢杆菌基因组DNA为模板,PCR扩增获得带有SD序列及谷氨酸棒状杆菌信号肽ΔS0949的BTG基因.将其与大肠杆菌-谷氨酸棒状杆菌穿梭表达载体pXMJ19连接,构建重组质粒pXMJ19-Sbtg转化谷氨酸棒状杆菌ATCC13032.经IPTG诱导后该重组菌发酵液具有交联酪蛋白的能力,表明该重组菌能够实现分泌表达.     

Biosynthesis of 1-alkenes in the defensive secretions ofTribolium confusum (Tenebrionidae); stereochemical implications

Summary The terminally unsaturated hydrocarbons of the defensive secretion ofTribolium confusum are biosynthesized from fatty acids by oxidative decarboxylation. The process involves an enantiospecific cleavage of the C–H bond of thepro-(S) C(3)–H atom and simultaneous decarboxylation of the acid into an 1-alkene and carbon dioxide via ananti-periplanar transition state geometry (anti-elimination). The stereochemistry of this biotranformation is identical in all respects with the same reaction in higher plants. The mechanism seems to be of general importance for the biosynthesis of many vinylic substructures of natural products from oxygen-containing precursors.     

Cholecystokinin is not a major regulator in the digestive system in the chicken

To find out whether physiological concentrations of cholecystokinin (CCK), a gastrointestinal hormone in mammals, are also active in chickens, the pancreatic amylase secretory response to CCK-8 was investigated in vitro. Rat pancreatic acini responded to the physiological concentration of CCK-8, but in chickens amylase release was induced at a concentration of CCK-8 1000 times higher than that observed in rats. In another experiment, biliary flow was tested with several doses of CCK-8. The bile flow was stimulated in a dose-dependent fashion, but a significant enhancement was not obtained at a concentration of 0.5 g CCK-8/kg body weight, which was considerably higher than physiological ones. It is concluded that endogeneous CCK does not have an important role in the digestive system in the chicken.     

Preliminary observations on the co-existence of regulatory peptides in cells of the baboon endocrine pancreas

Summary Immunocytochemical procedures at ultrastructural and light microscopy level revealed, in the Chacma baboon endocrine pancreas, cells which were immunoreactive for glucagon and pancreatic polypeptide (PP). Some D cells were observed to contain secretory granules with both the appearance and immunoreactivity of A cell secretory granules.     

枸杞多糖对Ⅱ型糖尿病小鼠胰岛细胞形态与功能的影响

采用高糖高热量饮食并结合小剂量腹腔注射STZ诱导Ⅱ型糖尿病（NIDDM）小鼠模型,枸杞多糖灌胃4周,用免疫组化技术对胰岛A、B细胞进行形态学分析及观察血糖、血清胰岛素的变化。结果显示,枸杞多糖大,小剂量组和模型组比较空腹血糖下降,胰岛素水平升高。图像分析大剂量组和模型组比较,B细胞数密度增加、A细胞数密度减小、B细胞切面积、核面积、核质比升高。结论：枸杞多糖可以改善Ⅱ型糖尿病小鼠胰岛细胞形态和功能并促进胰岛素的分泌,有良好的降血糖作用。     

胰腺腺泡细胞胞浆游离钙浓度的测定及药物作用的研究

本文建立了用荧光指示利Fura－2测定胰腺腺泡细胞胞浆游离Ca2＋浓度（［Ca2＋］i）的实验技术,包括：胰腺腺泡的分离制备;Fura—2／AM的负载;腺泡悬液的荧光光谱测量等．测定了胰腺腺泡细胞［Ca2＋］i。的静息值和卡巴可促腺泡细胞［Ca2＋］i升高的剂量—效应关系及动力学过程,并研究了柴胡总皂甙对胰腺腺泡细胞［Ca2＋i］的影响．发现0．01mg／ml浓度的柴胡总皂甙使胰腺腺泡细胞［Ca2＋］。明显升高,0．1mg／ml浓度时可使［Ca2＋］。升高到静息值的3倍,提示柴胡总皂甙的促酶分泌作用是通过胞内［Ca2＋］i升高介导的．     

人骨形态发生蛋白6(hBMP6)在毕赤酵母中的分泌表达

以重组质粒pcDNA6A-BMP6为模板PCR扩增hBMP6成熟肽cDNA编码序列,将该序列克隆到酵母分泌表达表达载体pPIC9K中.重组质粒pPIC9K-hBMP6经SacI酶切线性化,电击转化毕赤酵母GS115,PCR法筛选阳性转化子,利用梯度浓度G418筛选到七株高拷贝整合的阳性转化子。用甲醇进行诱导表达,SDSPAGE及Western-blot检测结果表明hBMP6成熟肽以分子量约为34和18 kD两种不同糖基化修饰的单体形式存在,具有良好的免疫原性.     

庆大霉素分泌过程的初步研究

应用恒温系统及缓冲液体系,考察了温度及pH值分泌过程的影响,然后在此基础上又讨论了添加葡萄糖、金属离子Fe^2 、K^ 和Mn^2 以及超声波和微波处理的效果,实验发现,pH＝7．4,温度34℃,以及添加5g／LKCl或50mg／LMnSO4都能刺激庆大霉素的分泌。     

柴胡皂甙促大鼠胰腺腺泡酶分泌作用的特征及胞外Ca2+对其的影响

利用检测胰腺腺泡蛋白分泌量表征胰酶分泌 ,研究柴胡总皂甙及柴胡皂甙单体 促胰腺腺泡酶分泌的量效特征和时效特征 ,并探讨胞外 Ca2 浓度的影响 .结果表明柴胡总皂甙及柴胡皂甙单体 促酶分泌作用随浓度增加而增强 ,且趋向饱和 .柴胡总皂甙促泌作用动力学呈现早期的高促分泌相和随后的低促分泌相 ;而 1 0 - 5 mol/L柴胡皂甙单体 的促泌动力学则出现了前期的低促分泌相 .胞外 Ca2 浓度为零 ,降低了柴胡总皂甙低促泌相的作用 ,并致使腺泡分泌趋向停止 .