血管周围脂肪组织源瘦素促进高脂喂食诱导的肥胖大鼠血管平滑肌细胞增殖

发布时间：2018-02-22 01:41

本文关键词： 肥胖血管周围脂肪组织瘦素血管平滑肌细胞　出处：《中华高血压杂志》2017年03期 　论文类型：期刊论文

【摘要】：目的探讨血管周围脂肪组织来源的瘦素对血管平滑肌细胞(VSMC)增殖的作用及其可能机制。方法 20只8周龄雄性Wistar大鼠随机分为2组,分别给予高脂饲料及标准大鼠饲料喂养,12周后筛选出7只肥胖大鼠作为肥胖组,普食组随机选取7只作为对照组,每周称重。喂养12周后,颈动脉插管测得血压;酶联免疫吸附试验(ELISA)检测大鼠血清瘦素水平;取大鼠降主动脉行HE及Masson染色,比较两组血管结构改变;Western blot法检测两组大鼠血管周围脂肪组织瘦素和瘦素受体蛋白表达水平。利用大鼠血管周围脂肪组织制备条件培养基,ELISA法检测条件培养基中瘦素水平;分别利用条件培养基、瘦素、瘦素拮抗剂(leptin-tA)、细胞外信号调节激酶(ERK)1/2抑制剂、p38抑制剂等干预大鼠VSMC,四甲基偶氮唑盐比色法(MTT)检测细胞增殖情况,Western blot法检测细胞瘦素、磷酸化ERK1/2(p-ERK1/2)、总ERK1/2(t-ERK1/2)、磷酸化p38(p-p38)、总p38(t-p38)蛋白表达。结果与对照组相比,肥胖组大鼠血清瘦素水平升高;主动脉壁结构紊乱,主动脉中膜厚度[(125.48±20.36)比(80.33±15.62)μm]和中膜厚度/管腔直径(0.14±0.01比0.08±0.01)增高(均P0.05)。大鼠血管周围脂肪组织瘦素及瘦素受体蛋白表达增高(均P0.05)。体外实验发现,条件培养基(血管周围脂肪组织细胞培养制成的上清液)瘦素水平明显升高[(8.34±0.92)比(4.38±0.66)μg/L,P0.05],且高于血清瘦素水平[(8.34±0.92)比(5.85±1.16)μg/L,P0.05];与对照组大鼠血管周围脂肪组织制备的条件培养基培养的VSMC相比,利用肥胖组大鼠血管周围脂肪组织制备的条件培养基培养的VSMC增殖显著增加,ERK1/2、p38磷酸化水平升高(均P0.05);加入瘦素拮抗剂后,细胞ERK1/2及p38磷酸化水平降低(均P0.05);加入瘦素拮抗剂、ERK1/2抑制剂、p38抑制剂后细胞增殖降低(P0.05)。结论血管周围脂肪组织来源的瘦素可以促进肥胖大鼠VSMC增殖,这种作用可能与ERK1/2和p38信号通路激活有关。
[Abstract]:Objective to investigate the effect of leptin derived from perivascular adipose tissue on proliferation of vascular smooth muscle cells (VSMC) and its possible mechanism. Methods Twenty 8-week-old male Wistar rats were randomly divided into two groups. Seven obese rats were selected as obese group after feeding with high fat diet and standard diet for 12 weeks, and 7 rats in general diet group were randomly selected as control group, weighing weekly. After 12 weeks of feeding, blood pressure was measured by carotid artery intubation. Enzyme linked immunosorbent assay (Elisa) was used to detect the serum leptin level in rats, and the descending aorta of rats was taken for HE and Masson staining. The expression levels of leptin and leptin receptor protein in perivascular adipose tissue of rats were detected by Western blot. Conditioned medium, leptin, leptin-tAn, extracellular signal-regulated kinase ERK1 / 2 inhibitor, p38 inhibitor, were used to interfere with VSMC in rats. The proliferation of VSMCs was detected by tetramethyl azolium colorimetry (MTT). The cell proliferation was detected by Western blot assay. Phosphorylated ERK1 / 2 / 2 p-ERK1 / 2, total ERK1 / 2t-ERK1 / 2, phosphorylated p38 + -p38, total p38-p38) protein expression. Results compared with the control group, the serum leptin level in obese rats increased, and the aortic wall structure was disordered. The thickness of medial membrane of aorta [125.48 卤20.36] was 80.33 卤15.62 渭 m, and the thickness of media / diameter of lumen was 0.14 卤0.01 vs 0.08 卤0.01 (all P 0.05). The expressions of leptin and leptin receptor protein in perivascular adipose tissue of rats were all increased (P0.05. The leptin level in conditioned medium (supernatant from perivascular adipocyte culture) was significantly increased [8.34 卤0.92 vs 4.38 卤0.66 渭 g / L], and higher than serum leptin level [8.34 卤0.92] vs 5.85 卤1.16 渭 g / L (P 0.05). The proliferation of VSMC cultured in conditioned culture medium prepared from perivascular adipose tissue of obese rats significantly increased the phosphorylation level of ERK1 / 2 p38 (all P 0.05). The levels of ERK1/2 and p38 phosphorylation were decreased (both P0.05%), and the proliferation of the cells was decreased after the addition of leptin antagonist (P 0.05). Conclusion leptin derived from perivascular adipose tissue can promote the proliferation of VSMC in obese rats. This action may be related to activation of ERK1/2 and p38 signaling pathways.
【作者单位】：西安交通大学医学院第一附属医院心血管内科;西安交通大学医学院第一附属医院超声科;陕西省人民医院心血管内三科;山东省立医院麻醉科;
【基金】：国家自然科学基金(30871042) 陕西省科学技术研究发展计划国际科技合作与交流计划项目(2012kw-40-01);陕西省科学技术研究发展计划自然科学基础研究计划项目(2014JM2-8145) 西安市科委攻关项目SF1416(2)
【分类号】：R544.1;R589.2

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