摘要
探讨氧化修饰的低密度脂蛋白（ＯＸ－ＬＤＬ）和氧化修饰的极低密度脂蛋白（ＯＸ－ＶＬＤＬ）是否能诱导巨噬细胞表达单核细胞趋化蛋白１（ＭＣＰ－１）ｍＲＮＡ和蛋白，以阐明二者在动脉粥样硬化发生中的作用。方法在巨噬细胞的培养基中分别加入２５μｇ／ｍｌ的低密度脂蛋白（ＬＤＬ）、极低密度脂蛋白（ＶＬＤＬ）、ＯＸ－ＬＤＬ和ＯＸ－ＶＬＤＬ，３７℃下温育２４小时，收集巨噬细胞的条件培养基，同时用异硫氰酸胍法提取巨噬细胞总ＲＮＡ。用Ｎｏｒｔｈｅｒｎｂｌｏｔ分析的方法检测巨噬细胞ＭＣＰ－１ｍＲＮＡ的表达，用夹心ＥＬＩＳＡ检测其条件培养基中ＭＣＰ－１蛋白的含量。用微孔滤膜法进行单核细胞的趋化试验。结果当巨噬细胞分别暴露于ＯＸ－ＬＤＬ和ＯＸ－ＶＬＤＬ２４小时后，其ＭＣＰ－１ｍＲＮＡ和蛋白的表达明显增加，同时单核细胞迁移距离的明显增加。而当巨噬细胞分别暴露于ＬＤＬ和ＶＬＤＬ２４小时，其ＭＣＰ－１ｍＲＮＡ和蛋白的表达只有轻微的增加，单核细胞迁移距离亦无明显的增加。结论巨噬细胞能表达ＭＣＰ－１ｍＲＮＡ和蛋白。
Objective To examine whether oxidatively modified low density lipoprotein(OX LDL)and verylow density lipoprotein(OX VLDL)induce the expression of MCP 1mRNA and protein by rabbit peritoneal exudate macrophages,and to clarify what arole of both lipoproteins play in atherogenesis.Methods After exposure of the macrophages to 25μg/ml of LDL,VLDL,OX LDL and OX VLDL respectively,and a 24 hour incubation at 37℃,the total RNA was extracted from the cells by guanidinium isothiocyanate method,and the media conditioned by the cultured macrophages were collected.Meanwhile,MCP 1protein in the conditioned media was determined by using sandwich ELISA.Monocy temigration induced by the media was assayed by micropore filter method using modified Boyden chamber.Results After a 24 hour exposure to OX LDL and OX VLDL,the MCP 1mRNA expression in macrophages was markedly increased(3.2 fold and 3.4 fold,respectively),and the level of MCP 1 protein was also increased(2.2 fold and 2.5 fold,separately),and furthermore,the monocyte migration distance was significantly increased.However,the expression of MCP 1mRNA and protein was only slightly increased when exposed to LDL or VLDL.Conclusions Rabbit peritoneal exudate macrophages can express MCP 1mRNA and protein,and OX LDL and OX VLDL induce strong erMCP 1mRNA and protein expression in the cells.
出处
《中华医学杂志》
CAS
CSCD
北大核心
1997年第3期212-215,共4页
National Medical Journal of China
基金
国家自然科学基金
