血卟啉单甲醚蛋白结合状态对其亚细胞分布的影响  

Influence of Combination Form of Hematoporphyrin Monomethyl Ether with Different Kinds of Serum Protein on Its Intracellular Distribution

作　　者：戴维德[1] 李晓松[1] 曾晶[1] 刘凡光[1] 顾瑛[1] 

机构地区：[1]解放军总医院激光医学科,北京市100853

出　　处：《中国激光医学杂志》2006年第3期137-140,共4页Chinese Journal of Laser Medicine & Surgery

基　　金：国家自然科学基金资助项目(60078021)

摘　　要：目的探讨在不同种类血浆蛋白的条件下血卟啉单甲醚向细胞内转运及分布的特点。方法在不同种类血浆蛋白条件下,将血卟啉单甲醚(HMME)与传代培养HeLa细胞共同孵育2h和12h。采用荧光探针标记技术和细胞器-细胞荧光强度比值分析法对细胞内HMME进行亚细胞定位和定量分析。结果孵育2h与12h比较,不含血清组中4种细胞器内HMME的平均荧光光强比值都有升高,以线粒体、内质网和高尔基体较显著,溶酶体稍有升高;白蛋白(Alb)组中4种细胞器中溶酶体的HMME平均荧光光强比值升高较显著,其他3种细胞器则略有升高;低密度脂蛋白(LDL)组中4种细胞器中溶酶体内HMME的平均荧光光强比值升高更加显著,其他细胞器的HMME平均荧光光强比值升高幅度明显大于Alb组。结论在短时间内进入细胞的主要为游离型HMME,转运形式以单纯被动扩散为主,以高尔基体、线粒体和内质网等细胞器为主要分布位点,溶酶体次之。随着孵育时间的延长,HMME可以和白蛋白、低密度脂蛋白结合形成结合型HMME,以胞吞形式进入细胞,二者相比,和LDL结合的HMME更易穿过细胞膜,溶酶体吸收HMME能力增强。Objective To discuss intracellular transportation and distribution of HMME in the situation of different kinds of serum proteins. Methods HeLa cells were subcultured and incubated with HMME for 2 and 12 hours in the situation of different kinds of serum proteins. Labeling technology of fluorescence probes and organelle-cell fluorescence intensity ratio analysis were adopted to study subcellular localization of HMME. Results Comparing cells incubated for 2h with that for 12h, the average fluorescence intensity ratio (J_1/J_2) of HMME of four organelles in group non-serum was all increasing, it was significant for three organelles including mitochondria, ER and Golgi; lysosome's J_1/J_2 of HMME in group albumin(Alb) was increasing more significantly than that of the other three organelles; among the four organelles in group LDL, lysosome's J_1/J_2 of HMME was increasing most significantly, in which the other organelles' increasing extents were apparently greater than that in group Alb.Conclusions At short time, HMME enters cells mainly in the form of dissociating HMME by passive diffusion and distributes in Golgi, mitochondria and ER,less in lysosome. With incubating time prolonging, HMME can be binded with albumin or LDL and enter cells by endocytosis, comparing with each other, HMME binding LDL can easily go through cell membrane than HMME binding albumin and is absorbed by lysosome.

关 键 词：血卟啉单甲醚 血浆蛋白 细胞内分布 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]