人体子宫内膜中性鞘糖脂和神经节苷脂的含量和组成在月经周期中和妊娠时的变化

从月经周期各期、早妊(6—8周)及足月妊娠子宫内膜中提取、纯化了神经节苷脂(Gls)和中性鞘糖脂(N-GSL),分别测定了其含量。对两类鞘糖脂的组成进行了HPTLC图谱分析。初步观察了不同时期子宫内膜中CMP—NeuAc:LacCer唾液酰转移酶(ST_1)和CMP-NeuAc:CM_3唾液酰转移酶(ST_2)的活性。结果表明:分泌期的GLs总含量低于增生期(P<0.02);分泌期GD_3含量较生长期增多(P<0.01);妊娠后,GM_3含量增加,而GD_3减少;相应地,ST_1活性增高,ST_2活性降低。分泌期CMH含量为增生期的4.7倍。结果提示子宫内膜鞘糖脂含量和组成的变化可能与子宫的功能有关,而且受女性激素水平的影响。     

妊娠期家兔子宫内膜的鞘糖脂表达

 妊娠期家兎子宫内膜的神经节苷脂(Gls)的含量明显低于动情期的,而中性鞘糖脂(NGSL)的含量则以妊娠中、晚期的最高,动情期最低。鞘糖脂组成变化最显著的是妊娠早期,由动情期到早孕GM_3从28.0％增加到52.7％,CMH.CDH由未测出分别增加到29.2％和21.9％,而糖链复杂的组分GD_3,GTlb和CPH的百分含量则明显减少,到妊娠中、晚期、短糖链组分逐渐减少,而复杂糖链组分渐增。中期妊娠内膜的(GIs)以GD_3为主要组分,占45％,明显高于其它各期。NGSL在妊娠中、晚期CPH增高达70％,与动情期水平相当。结果提示,妊娠期间子宫内膜的鞘糖脂含量与组成均发生明显变化,这些变化可能与子宫功能密切相关。特别是早孕对的变化,推测与子宫内膜和胚泡的识别,粘连特性的获得有关。     

雌、孕激素对去卵巢家兔子宫内膜鞘糖脂表达的影响

用雌、孕激素处理去卵巢家兔后观察子宫内膜鞘糖脂的含量和组成。结果表明、雌激素处理后、子宫内膜Gls含量是孕激素处理的十余倍,而孕激素增加NGSL含量的作用却比雌激素更强。两种激素对鞘糖脂组分的影响也明显不同。雌激素给药组子宫内膜鞘糖脂以多糖基组分GD_3 GT_(1b) CPH为主,孕激素组则以短糖链组分GM_3 CMH CDH为主。雌激素预先作用后再用孕激素或雌、孕激素同时给药,与单用雌激素后比较其短糖链组分明显增加,多糖基组分则明显减少,表现出两者的共同作用。上述结果提示:雌、孕激素对鞘糖脂代谢影响明显不同,雌激素使糖链复杂化、孕激素则使糖链趋向于简单。     

人子宫平滑肌肉瘤中性鞘糖脂含量、组成及其四糖基组分糖链结构的变化

本文分析了人子宫平滑肌肉瘤组织的中性鞘糖脂,发现子宫肉瘤组织的中性鞘糖脂的含量明显低于正常组织,组成成分虽与正常子宫平滑肌相似,均含有单、双、三、四糖基及多糖基组分,但各组分的相对含量则变化显著。肉瘤中所含短糖链的组分相对减少,高极性的CPH明显增加。本文还纯化了子宫肉瘤中性糖脂中主要的四糖基组分,应用HPTLC、酸解和酶解法以及特异单抗放射免疫染色法对该组分进行鉴定。结果证明子宫平滑肌中所含主要的四糖基组分糖链结构在正常组织为Globo系列的红细胞苷脂,而在肉瘤中则转变为乳糖系列的拟红细胞糖苷脂。     

自发性可移植性615小鼠肺癌和肝癌组织神经节苷脂高效薄层层析图谱的分析

本文研究了615近交系小鼠的自发性可移植性肺癌(P_(615))和肝癌(H_(615))组织中神经节苷脂结合唾液酸含量和神经节苷脂图谱。P_(615)和H_(615)癌组织中GLS结合唾液酸含量均比正常对照为高,分别为对照组织的2.9倍和1.9倍。615小鼠的正常肺组织和肝组织的GLS主要成分均为GM_3。在P_(615)和H_(615)癌组织中GM_3含量均明显减少。肺癌组织中GM_2大量增加,肝癌组织中不仅GM_2明显增加,GM_1和GD_(18)也明显增加。P_(615)和H_(615)这两种分化程度较高、恶性程度较低的癌组织GD_3的百分含量比正常对照组织略有降低。本文结果提示,自发性可移植性P_(615)和H_(615)肿瘤组织中不仅神经节苷脂含量(以唾液酸量计)升高,而且GLS的组分也发生改变。GM_3含量减少和GM_2含量增高可能与肿瘤的恶性生长和分化程度有关。     

狗小肠鞘氨醇糖脂中长链碱组成的研究

 利用气相色谱、气相色谱-质谱联用等方法测定了狗小肠鞘氨醇糖脂中的长链碱组成。其主要的长链碱为鞘氨醇(Sphingosine)、异鞘氨醇(isosphingosine)、二氢鞘氨醇(Sphinganine)和植物鞘氨酸(Phyto-sphingosine)。一共分离出十三个鞘氨醇糖脂。在唯一的五糖基神经酰胺中异鞘氨醇是主要成份。在一个一糖基神经酰胺中植物鞘氨醇是主要成份。植植物鞘氨酸也是两个二糖基神经酰胺和一个三糖基神酰胺的主要长链碱。说明它不仅存于植物体内。     

肝癌中CMP-乙酰神经氨酸:GM_3(α2→8)唾液酸转移酶(GD_3合成酶)的研究及纯化的初步报告

本实验室曾报道在所检测的不同种族来源与不同致癌剂所诱发的肝细胞肝癌中均有神经节苷脂GD_3组份的明显增高,本文就这一现象的机制进行了探讨。实验结果表明在人肝癌手术标本、人肝癌细胞株SMMC,3′-甲基奶油黄(3′Me-DAB)和二乙基亚硝胺(DENA)所诱发的大鼠肝癌以及大鼠肝癌株BERH-2中GD_3合成酶的活性均有不程度的增高,同对GD_3前体的合成酶(GM_3合成酶)的活性也有所增高。这就提示肝癌中GD_3增高的原因之一在于GD_3合成酶的活性增高与前体供应充足的结果。另外,本文还对GD_3合成酶的提纯做了初步尝试。主要采用Tritonx-100抽提和CDP-hydrazide Sepharose 4B亲合层析的方法从二乙基亚硝胺诱发的大鼠肝癌中提纯了GD_3合成酶。提纯倍数为12500倍,产率0.4％。提纯的GD_3合成酶在醋酸纤维膜上经等电聚焦电泳鉴定示单一条带,其pI值为5.25左右。关于糖脂唾液酸转移酶的纯化工作目前还未见报道。     

高效液相色谱(HPLC)技术鉴定中国南方大豆品种异黄酮主要组分

采用高效液相色谱(HPLC)技术检测中国南方六个省份的249份大豆品种异黄酮主要组分含量.结果显示大豆籽粒中可检测出6种主要的异黄酮组分,分别为大豆甙(Daidzin)、甲氧基黄豆甙原(Glycitin)、染料木甙(Genistin)、丙二酰基大豆甙(Malonyldaidzin)、丙二酰基黄豆甙原(Malonylglycitin)和丙二酰基染料木甙(Malonylgenistin).各组分中以丙二酰基(Malonyl)异黄酮组分含量最高(61.2%),且各组分间相关极显著.大豆品种间异黄酮含量变异较大,变异系数达49.6%.来自江苏省的品种海门红黄豆乙异黄酮含量最高(4932.3μg/g),品种宝应等西风含量最低(367.1μg/g).不同省份间异黄酮含量差异极显著,来自浙江省的大豆品种平均含量最高(2717.2μg/g),来自安徽省的平均含量最低(1181.8μg/g).异黄酮含量与生育期呈极显著正相关(r=0.319* * *),与百粒重呈显著正相关(r=0.132*),而与脂肪含量(r=-0.45* * *)和蛋白质含量(r=-0.136)呈负相关.     

GM_3在几种哺乳动物肝脏和狗红细胞中的含量及其分离纯化

从正常兔、猪和狗的肝脏及狗红细胞中分离纯化了总神经节苷脂,测定了脂结合唾液酸,进行了高效薄层层析,比较了上述四种组织中GM_3的含量。结果表明狗红细胞中的GM_3的含量较另三种的高,狗肝和兔肝次之,猪肝含量甚微。从狗红细胞中提取和纯化了GM_3,其得量为每毫升压积红细胞351.0μg,纯度为92.2％。     

基于糖显色法测定鼠李糖脂的比例、含量

目的:确定微生物培养液中鼠李糖脂的组成、含量和鼠李糖脂在电喷雾质谱(ESI-MS)中的离子化效率.方法:用薄层色谱分离并结合ESI-MS分析培养液中的糖脂产物、测定样品中单鼠李糖脂和双鼠李糖脂比例及离子化强度.结果:根据苯酚-硫酸法和蒽酮光度法中糖及糖脂与吸光度的定量关系[A=0.0103X+0.0465(X为鼠李糖量,μg),A=0.0043X+0.0446(X为鼠李糖脂量,μg)],用化学计量方法确定了糖脂含量84.8%,其中单鼠李糖脂的质量分数0.344,双鼠李糖脂的质量分数0.656,并基于电喷雾质谱中的离子强度和测定的浓度计算了鼠李糖脂的离子化效率,双鼠李糖脂的钠离子化效率仅为单鼠李糖脂钠离子化效率的50%.结论:可用于定量分析单双糖脂及评价鼠李糖脂的生产.     

舟山眼镜蛇毒抗肿瘤有效组分分离及其抑瘤作用研究初探

目的从舟山眼镜蛇(Naja atraCantor)蛇毒(snake venom,SV)中分离得蛇毒组分,探讨SV及其分离组分的LD50和抑制肿瘤的作用。方法采用凝胶柱层析方法从蛇毒中分离得到了前Ⅰ1、Ⅰ1、Ⅱ1、Ⅱ2、Ⅲ1、Ⅲ2及Ⅳ等7种组分。采用急性毒性实验、MTT法,研究SV及其7种SV分离组分的LD50和抑制肿瘤的作用。结果 SV经Sephadex G-50层析,可分离为前Ⅰ、Ⅰ、Ⅱ、Ⅲ及Ⅳ5个组分,根据峰面积大小排列:ⅢⅡⅠⅣ前Ⅰ。5个组分再经Sephadex G-25柱层析,可获得7个脱盐组分:前Ⅰ1、Ⅰ1、Ⅱ1、Ⅱ2、Ⅲ1、Ⅲ2及Ⅳ。通过急性毒性实验,明确Ⅳ的毒性最大,其次为Ⅲ2及Ⅲ1,三者的LD50值均低于SV;而Ⅰ1、Ⅱ1、Ⅱ2的毒性均小于SV,前Ⅰ1几乎无毒。SV组分Ⅲ2和Ⅳ的抑瘤作用最强,在高浓度(20μg/ml)时对实验中的2种人肿瘤细胞的抑制率均达到60%以上。结论从SV中分离得到了前Ⅰ1、Ⅰ1、Ⅱ1、Ⅱ2、Ⅲ1、Ⅲ2以及Ⅳ7种组分;组分Ⅳ毒性最强,依次为Ⅲ2Ⅲ1SVⅡ2Ⅱ1Ⅰ1前Ⅰ1;SV及其7种分离组分对2种人肿瘤细胞株(SGC-7901、A549)的生长抑制有一定的特异性,而不同的SV分离组分对同一肿瘤细胞抑制作用亦有差异。     

Glycosphingolipids of human myometrium and endometrium and their changes during the menstrual cycle, pregnancy and ageing

The glycolipid composition of human myometrium and endometrium was examined at various stages of maturation and reproduction. The major neutral glycolipids of both myometrium and endometrium were identified by high-performance thin-layer chromatography as globo-series glycolipids, Gb3 and Gb4. The major acidic glycolipids (gangliosides) were identified similarly as GM3 and GD3, with lesser amounts of GM1, GD1a, and GT1b. During pregnancy, GD3 expression declined in both myometrium and endometrium, whereas GM3 expression increased. Reciprocal changes in GM3/GD3 expression were mirrored by appropriate changes in the glycosyltransferases required for their synthesis; alpha 2----3sialyltransferase activity increased approximately 3-fold during pregnancy, while alpha 2----8sialyltransferase activity declined to about 20%. The results focus attention on the glycolipids of uterine tissues, their regulation, and their possible role in reproduction and fertility.     

Specific gangliosides increase rapidly in rat liver following partial hepatectomy

Rat liver gangliosides (sialic acid containing glycosphingolipids) were analyzed by HPTLC and HPLC following either partial hepatectomy or sham operation. Analysis of whole liver gangliosides by HPTLC demonstrated that within 6 h after partial (68%) hepatectomy, there was a significant increase in GM1 compared to both sham and control animals. By 48 h, GM1 was further increased and the polysialylgangliosides GD1a, GD1b and GT1b had also risen significantly, whereas changes in GM3 were negligible. Gangliosides associated with the plasma membrane were increased up to 3.5-fold in regenerating liver compared to sham-hepatectomized controls as assessed by HPLC. Although elevations in membrane gangliosides were associated with hepatocyte proliferation, they did not closely follow the growth curve. The time course of changes in ganglioside biosynthesis suggests differential upregulation of GM3 synthase and GD3 synthase in regenerating livers.     

Molecular parameters and physical state of neutral glycosphingolipids and gangliosides in monolayers at different temperatures

The effect of temperature on the behaviour of four different gangliosides (GM3, GM1, GD1a and GT1b), sulphatide, ceramide (Cer) and three neutral glycosphingolipids (GalCer, Gg3Cer, Gg4Cer) was investigated in monolayers at the air-NaCl (145 mM) interface. GM1, GD1a and GT1b are liquid-expanded in the range of temperatures studied (5-65 degrees C). GM3, sulphatide, Cer and neutral glycosphingolipids show isothermal liquid-expanded----liquid-condensed transitions. The collapse pressure of ganglioside monolayers decreases with temperature, whereas neutral glycosphingolipids may show some maximum values at particular temperatures. The reduction of the molecular area of liquid-expanded glycosphingolipids under compression occurs with a favorable positive entropy change and an unfavorable negative enthalpy. By contrast, the compression of interfaces with a two-dimensional phase transition occurs with an unfavorable entropy but a favorable enthalpy change. From the temperature dependence of the surface pressure at which the two-dimensional phase transition takes place, a minimal temperature above which the isotherm becomes totally liquid-expanded can be obtained. For the different glycosphingolipids this temperature decreases in the order Cer greater than GalCer greater than sulphatide greater than Gg3Cer greater than Gg4Cer greater than GM3 greater than GM1 greater than GD1a greater than GT1b. This sequence is similar to that found for the calorimetrically determined transition temperatures (cf. Maggio, B., Ariga, T., Sturtevant, J.M. and Yu, R.K. (1985) Biochemistry 24, 1084-1092).     

Surface plasmon resonance study of carbohydrate-carbohydrate interaction between various gangliosides and Gg3-carrying polystyrene

Carbohydrate-carbohydrate interactions between Gg3 trisaccharide-carrying polystyrene (PN(Gg3)) and monolayers of several glycosphingolipids (GSLs) were quantitatively investigated by surface plasmon resonance techniques. PN(Gg3) was adsorbed onto a GM3 monolayer strongly and specifically with an apparent affinity constant of K(a) = 2.5 x 10(6) M(-1), and the apparent affinity constants onto GSLs decreased in the following order: GM3 > LacCer > (KDN)GM3 approximately GlcCer > GM2 approximately GD3 approximately GM4 > GM1 approximately 2,6-isoGM3 > ceramide. These results suggest that PN(Gg3) recognizes not only some specified portions of GM3 but also the trisaccharide as a whole. On the other hand, PN(Lac) and PN(Cel) were bound to GSLs less strongly (K(a) approximately 10(4) M(-1)) and less selectively. The kinetic analysis revealed that the selectivity in the adsorption of PN(Gg3) onto the GM3 monolayer is dominated by the faster adsorption rate.     

手机辐射对孕鼠及子代IFN-γ和IL-4的影响

检测暴露于手机辐射下的孕鼠及其子代的细胞免疫因子含量水平,以探讨手机电磁辐射对孕鼠及其子代免疫功能的影响,为孕妇科学、合理使用移动通信工具提供参考。30只孕鼠随机分为Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ组,每组6只,对应为空白对照组、待机对照组1、0min低强度组、30min中强度组和60min高强度组。ELISA检测孕鼠分娩24h内孕鼠及新生乳鼠外周血中IFN-γ和IL-4的含量。对Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ各组母鼠外周血中IFN-γI、L-4含量进行比较,其差异无统计学意义(P>0.05);Ⅳ、Ⅴ组新生小鼠外周血中IFN-γI、L-4含量与Ⅰ、Ⅱ组比较,其差异有统计学意义(P<0.05),而且Ⅴ组新生小鼠外周血中IFN-γI、L-4含量与Ⅲ组比较,其差异也具有统计学意义(P<0.05)。手机辐射降低了胚胎期小鼠的IFN-γ和IL-4含量。     

Ganglioside composition of astrocytes

Short-term and long-term (greater than 7 months) cultured astrocytes from 14-day-old rat brain were analyzed for ganglioside content. Analysis of the extracted gangliosides by HPTLC revealed that ganglioside GM1 was absent in 35 days and 235 days cultured astrocytes, and that the predominant ganglioside was GM3, showing a double band in both cases. A small amount of the disialogangliosides (GD3, GD1a) was also detected. More than 70% of radioactivities into ganglioside fractions by cultured astrocytes, in the presence of N-[3H]-acetylmannosamine, appeared in ganglioside GM3. The upper band component of GM3 increased 60% in long-term astrocyte cultures compared to 35-day-old cultures. Also, an increased GD3 content in long-term astrocyte cultures was detected. These results suggest that the increase of GD3 and upper band GM3 in long-term cultured astrocytes might be related to the appearance of small processes showing strong reactivity against GFAP and vimentin during astrocyte-subculture.     

猪脑中提取高纯度神经节苷脂

应用凝胶层析和离心液相层析法,从猪脑中提取高纯度神经节苷脂(Gls).按脂结合唾液酸(LBSA)计为30.1%,经硅胶薄层层析,580nm扫描结果表明含5种Gls,即:GM1为19.5%,GD3为13.8%,GD1a为27.8%,GD1b为14.2%和GT1b为19.3%.