摘要
目的探讨同型半胱氨酸(Hcy)是否通过p38丝裂原活化蛋白激酶(p38 MAPK)通路诱导大鼠主动脉平滑肌细胞表达依赖正常T细胞激活和分泌的因子(RANTES)和其趋化因子受体1(CCR1)及辛伐他汀的治疗作用。方法将26只健康8周龄SD雄性大鼠随机分为正常对照组、高蛋氨酸饮食组和辛伐他汀治疗组。正常对照组给予普通饲料喂养;高蛋氨酸饮食组以普通饲料中添加质量分数2%的蛋氨酸喂养;辛伐他汀治疗组以高蛋氨酸饮食组饲料喂养4周后,每天以5 mg.kg-1的辛伐他汀灌胃,并继续给予高蛋氨酸饮食组饲料。12周后采用高效液相色谱法检测血浆Hcy浓度,苏木素-伊红(HE)和油红"O"染色观察胸主动脉形态学变化,采用免疫组织化学和逆转录聚合酶链式反应(RT-PCR)检测大鼠胸主动脉平滑肌细胞中RANTES、CCR1蛋白和mRNA的表达;采用Western blotting检测大鼠胸主脉平滑肌细胞中p38 MAPK和磷酸化p38 MAPK(p-p38 MAPK)蛋白的表达。结果蛋氨酸饮食组血浆Hcy浓度(80.79±23.86)mmol.L-1高于正常对照组(9.47±4.55)mmol.L-1和辛伐他汀治疗组(53.94±12.24)mmol.L-1(P<0.01)。HE染色显示高蛋氨酸饮食组局部内皮细胞坏死脱落,内膜下可见泡沫细胞。油红"O"染色显示高蛋氨酸饮食组胸主动脉内膜下局部呈红色。高蛋氨酸饮食组RANTES蛋白表达的平均灰度值(100.54±15.31)低于正常对照组(114.25±10.73)(P<0.01)和辛伐他汀治疗组(113.12±10.86)(P<0.05)。高蛋氨酸饮食组CCR1蛋白表达的平均灰度值(120.12±7.71)低于正常对照组(124.31±9.44)(P<0.01)和辛伐他汀治疗组(123.62±8.32)(P<0.05)。高蛋氨酸饮食组大鼠胸主动脉平滑肌细胞中RANTES和CCR1 mRNA的表达分别为0.93±0.02、1.77±0.40,均高于其在正常对照组的表达(0.45±0.00、1.02±0.04)(P<0.01),并高于在辛伐他汀治疗组中的表达(0.69±0.01、1.22±0.53)(P<0.05)。p38 MAPK在各组大鼠动脉平滑肌细胞均有表达但差别无统计学意义(P>
Objective To investigate whether homocysteine（Hcy） induced the expressions of regulated upon activation normal T expression and secretion（RANTES） and its receptor chemoattractant cytokine receptor-1（CCR1）through p38 mitogen activated protein kinase（p38 MAPK） pathway in the arterial smooth muscle cells of the rats.Methods Twenty-six of healthy 8-week-old SD male rats were randomly divided into normal control group,high methionine diet group and simvastatin treatment group.The rats of normal control group were fed with normal food,and the rats of high methionine diet group were raised with food including normal food and 2% methionine.The rats of simvastatin treatment group were fed with high methionine group diet for four weeks,then they were intragastric administrated with simvastatin at 5 mg·kg-1 everyday,and fed with high methionine group diet.After 12 weeks,the concentrations of plasma Hcy were detected by high performance liquid chromatography（HPLC）.The morphological alterations of the thoracic aorta were determined by hemotoxylin and eosin（HE） staining and oil red ＂O＂ staining.The expressions of RANTES and CCR1 proteins and mRNA were detected with immunohistochemistry and reverse transcription polymerase chain reaction（RT-PCR）.The expressions of p38 MAPK and phosphorylated p38 mitogen activated protein kinase（p-p38 MAPK） proteins were detected with Western blotting.Results The concentration of plasma Hcy in high methionine diet group[（80.79±23.86）mmol·L^-1] were significantly higher than those in the normal control group[（9.47±4.55）mmol·L^-1] and simvastatin treatment group[（53.94±12.24）mmol·L^-1]（P〈0.01）.HE staining showed some endothelial cells were detached,and there were foam cells under the endothelium of the thoracic aorta in the high methionine diet group.The oil ＂O＂ staining showed some local areas of the thoracic aorta were stained positively with red.Immunohistochemistry showed that the average grey value of RANTES protein in the high
出处
《新乡医学院学报》
CAS
2011年第6期703-707,710,共6页
Journal of Xinxiang Medical University
