热应激干预诱导产生HSP70对大鼠动脉粥样硬化的影响

目的 通过施加热应激干预,检测大鼠血清血脂、热休克蛋白70(heat shock protein 70,HSP70)、活性氧(radical oxygen species,ROS)、氧化低密度脂蛋白(oxidized low density lipoprotein,ox-LDL)的表达水平,探讨热应激干预处理对大鼠动脉粥样硬化的影响,为临床防治心脑血管疾病提供基础理论依据。 方法 将30只SPF级雄性SD大鼠随机分为Control组、AS组、H+AS组(n_i=10)。Control组予基础饲料饮食;AS组实验开始第1天一次性维生素D₃(6×105U/kg)腹腔注射给药,每隔30 d给药1次,共给药3次+高脂饲料饮食进行AS动物造模;H+AS组于AS动物造模期间,隔天1次,施加热应激干预至造模时间结束。14周后颈总动脉取血,ELISA法检测血脂、HSP70、ox-LDL,Fenton法测定ROS以及观察动脉血管病理改变。 结果 采用单因素方差分析,与Control组比较,AS组血清TC(31.87±3.79) nmol/L vs.(14.14±2.95) nmol/L]、TG(226.00±17.09)μmol/L vs.(106.10±17.57)μmol/L]、LDL(169.17±18.75)μmol/L vs.(96.55±13.92)μmol/L]、ox-LDL(26.68±3.52)μg/L vs.(9.92±3.10)μg/L]明显升高(P<0.01),病理示动脉内膜增厚,可见泡沫细胞,典型粥样斑块形成;与AS组比较,H+AS组血清TC(28.30±2.99) nmol/L vs.(31.87±3.79) nmol/L]降低,差别具有统计学意义(P<0.05),TG(168.68±26.17)μmol/L vs.(226.00±17.09)μmol/L]、LDL(137.47±16.30)μmol/L vs.(169.17±18.75)μmol/L]明显降低(P<0.01),HSP70(673.39±130.93) ng/L vs.(324.96±57.34) ng/L]明显升高(P<0.01),但是ROS水平也升高,差别具有统计学意义(P<0.01),病理示动脉内膜钙盐沉积,但未见典型AS斑块形成。 结论 热应激干预的方式在表观上具有抑制大鼠动脉粥样斑块形成的效果,可能与诱导产生的HSP70在发病过程中的保护作用有关。 

Effects of HSP70 induced by heat stress intervention on atherosclerosis in rats

Objective To provide basic theoretical basis of atherosclerosis for the prevention and treatment of cardiovascular and cerebrovascular diseases by detecting the expression level of serum lipids, heat shock protein 70 (HSP70), radical oxygen speciesn (ROS) and oxidized low density lipoprotein (ox-LDL) in rat with heat stress pretreatment. Methods SD rats were randomly divided into control, the AS and H + AS groups. The control group with based diet, the AS and H + AS were treated with intraperitoneal injection a single dose of vitamin D₃ (6×105 U/kg) in the first days of experiment began. Then, every 30 days administration of 1 times, a total of three times administration in the next time combines with high fat diet. During the AS animal modeling, the H + AS group was pretreated with heat stress intervention in every one days until the molding time end. After 14 weeks, draw blood from the common carotid artery, centrifuge, HSP70, ox-LDL and blood lipid were detected by ELISA method, ROS was detected by Fenton method and observe the arterial pathological changes. Results By one-way analysis of variance, in AS group, compared with control group, the levels of TC(31.87 ±3.79) nmol/L vs. (14.14 ±2.95) nmol/L], TG(226.00 ±17.09) μmol/L vs. (106.10 ±17.57) μmol/L], LDL(169.17 ±18.75) μmol/L vs. (96.55 ±13.92) μmol/L], ox-LDL(26.68 ±3.52) μg/L vs. (9.92 ±3.10) μg/L] increase (P<0.05), the arrangement of endothelial cells was in disorder, foam cells were observed and typical atherosclerosis developed. In H + AS group, compared with AS group, the levels of TC(28.30 ±2.99) mmol/L vs. (31.87 ±3.79) mmol/L] decreased (P<0.05), TG(168.68 ±26.17) μmol/L vs. (226.00 ±17.09) μmol/L] and LDL(137.47 ±16.30) μmol/L vs. (169.17 ±18.75) μmol/L] decreased (P<0.01), HSP70(673.39 ±130.93) ng/L vs. (324.96 ±57.34) ng/L]increased significantly (P<0.01), but ROS is also increased (P<0.01), calcium salts accumulated in the arterial endarterium, but typical atherosclerosis plaques were barely found. Conclusion The method of heat stress intervention can inhibit the formation of atherosclerotic plaque in rats which may be related to the protective effect of HSP70 in the pathogenesis of the disease.