质膜流动性与细胞分裂

质膜流动性与细胞分裂王英杰（杭州大学生物科学与技术系,杭州３１００１２）董萍（浙江省化工研究院,杭州３１００２３）关键词质膜,膜流动性,细胞分裂自从１９７２年Ｓｉｎｇｅｒ和Ｎｉｃｏｌｓｏｎ提出生物膜的“流动镶嵌模型”以来,大量的实验证据已使我们对生物...     

脂肪酸对人肺腺癌细胞膜流动性的影响

脂肪酸是细胞膜正常流动性的主要调节因素之一。本文报导了二种不同转移表型人肺腺细胞与九种不同脂肪酸共孵育后,对其细胞膜流动性的影响。结果表明,不同转移一夫肺腺癌细胞对各种脂肪酸有不同的敏感性,高转移癌细胞Ａｎｉｐ对棕榈酸和花生酸较敏感,而低转移癌细胞ＡＧＺＹ对棕榈烯酸和亚油酸较敏感。     

HL-60细胞诱导分化与细胞膜流动性变化研究

 本实验以二甲基亚砜(Dimethyl Sulfoxide,DMSO)为诱导剂,诱导人早幼粒白血病细胞系HL-60沿粒系统成熟分化。动态观察了诱导后1—6天HL-60的形态、功能成熟度改变、膜流动性和增殖活性的变化。结果表明,早幼粒细胞由诱导前的78％降至5％,杆核与分叶核细胞由6％和1.5％分别增加至32.5％和5.5％。NBT(Nitroblue tetrazolium,四唑氮蓝)还原试验显示其功能亦渐超成熟。诱导后HL-60 DNA合成速率明显降低(降低40～75％),膜脂流动度亦显著降低(降低30％,P<0.01)。提示诱导分化后的HL-60细胞不仅获得了与正常成熟粒细胞相似的形态功能特征,同时还失去了膜流动性增高及快速繁殖增生的恶性特征。     

RFP134对UMR106细胞膜脂流动性的影响

癌细胞具有与正常细胞不同的膜脂流动性,导致细胞对生长因子和癌基因产物反应敏感,引起细胞增殖失控。本实验室从植物中发现一种二萜类活性物质－RFP134,在细胞周期和信号传递等多方面表现出有抑制癌细胞增殖,促进细胞分化的作用。本文以大鼠成骨肉瘤细胞（UMR106）和下鼠成骨细胞为模型,研究其对癌细胞膜脂流动性的影响。细胞系UMR106由美国麻省总医院内分泌室赠送。成骨细胞由本实验室分离培养。以不同浓度（20、40、60、80、100μM／L）的RFP134,在同一时间处理细胞,或以最适浓度（50μM／L）在不同时间作用于细胞。DH为荧光标记物,测得的荧光偏值和微粘度值为膜膜流动性指标。结果显示,无论在恒定的时间、以不同浓度的RFP134作用于UMR106细胞(Fig.1B）,或以恒定的浓度、在不同时间处理UMR106细胞（Fig.1D）,结果均表现为显著降低膜脂流动性。前者,RFP134作用于细胞时,细胞荧光偏振瑟微粘度值逐步升高,其变化呈量效关系;而后者,呈时效关系。但在最适浓度与最佳作用时间,荧光偏振值和微粘度值达饱和状态。在同样条件下,RFP134对正常成骨细胞的膜脂流动性影响小。即：荧光偏振值和微粘度值均在正常范围内保持恒定（Fig.1A;Fig.1C）RFP134降低癌细胞的膜脂流动性,从而改善了它的细胞膜功能,降低了它对生长因子的反应性,恢复了细胞对调节因子的正常反应。这可能是RFP134能够抑制癌细胞增殖,促进细胞分化的机制之一。     

脂质过氧化对人红细胞膜脂流动性的影响

研究枯稀过氧化氢/高铁血红素体系所产生的烷基过氧自由基对红细胞的损伤。测定了脂质过氧化的产物——丙二脂的生成,并证明阿魏酸钠对脂质过氧化的抑制。荧光偏振的结果指出,膜脂过氧化以后降低了膜脂的流动性。人红细胞用5DSA和16DSA标记并用ESR检测膜脂流动性,结果表明,序参数S几乎没有发生变化,旋转相关时间τ值的增加证明膜脂过氧化以后,疏水尾部的物理状态发生了改变。经脂质过氧化以后,红细胞膜中的不饱和脂防酸的减少,可能是降低膜脂流动性的原因之一。     

单纯疱疹病毒感染细胞膜系统脂质分子流动性的变化

运用荧光偏振技术了解单纯疱疹病毒(HSV)I型Sm44株感染非洲绿猴肾(Vero)细胞后细胞膜系统脂质分子流动性的变化.结果表明,在所观察的感染后的5个时相中(感染后0、7、14、21、28小时),感染早期(感染后0、7小时)细胞膜的流动性变化不大;感染晚期(14至28小时)细胞膜的脂质流动性明显增强,与未感染病毒的对照组细胞有显著差异,这提示HSV感染晚期的装配,成熟与释放是与被感染细胞的生物膜系统密切相关的.本文对运用该生物物理技术研究病毒和宿主细胞相互关系的意义进行了初步讨论.     

不同剂量的He—Ne激光对Raji细胞膜脂流动性的影响

本文采用荧光偏振法检测了不同剂量的Ｈｅ－Ｎｅ激光照射法对Ｒａｊｉ细胞膜流动性的影响,发现了０．０１Ｊ／ｃｍ＾２和０．５１Ｊ／ｃｍ＾２两剂量可使膜脂微粘度降低（Ｐ〈０．０５）。即其膜脂流动性升高;１．０Ｊ／ｃｍ＾２和２．０Ｊ／ｃｍ＾２的剂量脂微粘度无明显影响,即其膜流动性无明显化（Ｐ〉０．０５）,而６．０２Ｊ／ｃｍ＾２的剂量可使膜微粘度明显上升（Ｐ〈０．０５）。即其膜流动性明显下降。     

犬实验性急性心肌梗塞早期心肌细胞膜脂流动性的改变

为了解急性心肌梗塞(AMI)时心肌细胞膜脂流动性(LFU)的变化,用荧光分光光度计偏振法测定了20只实验性AMI早期犬的心肌细胞LFU。结果表明,AMI早期心肌细胞LFU与正常组比较明显降低(P<0.01),其降低程度与心肌缺血时间长短和心肌细胞损伤程度有关。LFU在冠状动脉(CA)结扎后1min时即出现降低,与心外膜电图S—T段弓背向上损伤性改变同时发生,而明显早于外周血清肌酸磷酸激酶(CPK)和谷草转氨酶(AST)以及心肌组织病理形态学的改变,后者在CA结扎后240min出现变化。因此,LFU可作为AMI早期反映心肌细胞缺血损伤及其程度的一种敏感标志。     

巨噬细胞激活及钙作用的初步研究

经TG诱发的小鼠腹腔渗出液的巨噬细胞及人的巨噬细胞样细胞株U937受PAF(100ng ml).Zymosan A(0.25mg ml).Can A(50μg ml)LPS(1μg ml)等作用后.能引起胞内游离Ca~(2-)浓度的增加,巨噬细胞内酸性磷酸酶增多,细胞骨架更为舒展、丰满.胞内游离Ca~(2-)的增加是由于胞内钙库的释放与胞外钙的内流.上述困子作用后,可使巨噬细胞产生呼吸爆发.其中.Zymosan A的作用尤为强烈.同时还出现胞膜流动性的降低、当胞外环境中有Ca~(2-)时.可增强巨噬细胞的呼吸爆发.以上提示:在巨噬细胞的激活中Ca~(2-)具有重要的作用.     

Cheek cell membrane fluidity measured by fluorescence recovery after photobleaching and steady-state fluorescence anisotropy

Membrane fluidity of human cheek cells was determined using fluorescence recovery after photobleaching (FRAP) and steady-state fluorescence anisotropy. The FRAP data showed that the lateral diffusion coefficient (D) and mobile fraction (%R) of lipid in the plasma membrane of control cells were 2.01×10^–9 cm²/ sec and 54.25%, respectively. Trypsin treatment increased D and %R to 6.4×10^–9 cm²/sec and 72.15%. In contrast, the anisotropy (r) for control cells was 0.270 which remained unchanged by trypsin treatment. The results show that diffusion of lipids in the plane of the membrane is restricted by trypsin-sensitive barriers.     

Selective accumulation of hybrid liposomes into adult T-cell leukemia cells along with induction of apoptosis

Markedly inhibitory effects of hybrid liposomes (HL-n) composed of 90 mol % l-α-dimyristoylphosphatidylcholine (DMPC) and 10 mol % polyoxyethylene(n) dodecyl ethers on the growth of adult T-cell leukemia cells were obtained for the first time. It is noteworthy that HL-n could selectively accumulate into the adult T-cell leukemia cells and induce apoptosis via caspase-3 activation.     

用ANS研究脱落酸提高植物线粒体膜的流动性

用荧光标记法,以ＡＮＳ作荧光探剂研究了脱落酸（ＡＢＡ）对植被线粒体脱流动性的影响。（＋）ＡＢＡ浓度效应实验结果和不同ＰＨ及不同温度条件下的实验结果均表明（＋）ＡＢＡ能降低植物线粒体膜的荧光强度,即（＋）ＡＢＡ能提高线粒体原流动性,且此种效应具有浓度饱和性,并导致线粒体异柠檬酸脱氢酶（ＩＣＨ）活性升高。ＡＢＡ的类似物,ＲＣＡ．７Ａ和ＲＣＡ．７ｂ,也具有提高ＩＣＤＨ活性的效应,抗ＡＢＡ结合蛋白的抗体（     

中华眼镜蛇毒心脏毒素对人肝癌细胞株细胞膜的影响

目的和方法：运用５７０ 型粘附式细胞仪光漂白后荧光再分布法测定单个细胞膜脂流动性的动态变化和流式细胞仪测定细胞群细胞膜电位,以观察中华眼镜蛇毒心脏毒素对人肝癌细胞株细胞膜的影响。结果：心脏毒素使肝癌细胞膜脂流动性下降,并且使肝癌细胞膜电位下降。结论：心脏毒素对人肝癌细胞株Ｈ７４０２细胞膜有损伤作用     

Differential effect of triiodothyronine and thyroxine on the liposomal membrane in liquid-crystalline and gel state

The effect of thyroid hormones on the degree of order or fluidity of dimyristoyl, dipalmitoyl or egg yolk phosphatidyl choline liposomes was evaluated by fluorescence spectroscopy methods. The freedom of molecular motion above the phase transition temperature was decreased, while below the transition, the mobility was actually increased by the incorporation of triiodothyronine to liposomes. While thyroxine decreases the fluidity in the liquid crystalline state, it cannot increase the fluidity in the gel state.A differential effect of triiodothyronine and thyroxine on the release of the liposomal content was found, depending on the liquid crystalline or gel state of the liposomes. These facts were correlated with the differential incorporation of the hormones to liposomes above and below the phase transition temperature of dimyristoyl and dipalmitoyl phospholipid choline. In gel state, a low incorporation of thyroxine compared with triiodothyronine was found.This work was supported by Grants PID 3-013800/89 from Consejo National de Investigaciones Científicas y Técnicas (CONICET), Fundación Antorchas A-12576/1-000065 and Consejo de Investigaciones de la Universidad National de Tucumán (CIUNT). We thank Dr. G. Rotillo for the space filling models.     

Incorporation and fate of specific sperm plasma membrane components following insemination as revealed by ultrastructural immunocytochemistry

Studies have been carried out to 1) further characterize sperm specific plasma membrane polypeptides (33 and 35 kDa) that are recognized by a monoclonal antibody previously described (Longo, 1989) and 2) follow the incorporation and dispersal of these proteins within plasmalemmae of monospermic and polyspermic sea urchin (Arbacia punctuluta) eggs and oocytes utilizing immunocytochemical methods at the ultrastructural level of observation. Only sperm labeled when incubated with monoclonal antibody to the 33 and 35 kDa proteins followed by colloidal gold-tagged second antibody. Colloidal gold label was observed on the egg plasma membrane immediately after gamete membrane fusion; the amount and extent of label, i.e., the distance from the site of sperm incorporation, increased with time postinsemination. By 20 min postinsemination approximately one hemisphere of the inseminated egg/oocyte was associated with label. The expanding distribution of colloidal gold label on inseminated eggs and oocytes vs. time reflects the free diffusion of 33 and 35 kDa sperm surface proteins among egg/oocyte plasma membrane components. Label was also found in forming endocytotic vesicles, suggesting that sperm plasma membrane proteins may be internalized.