多肽化合物urantide对动脉粥样硬化大鼠胸主动脉和VSMC中Ⅳ型胶原表达的影响

doi:10.13481/j.1671-587x.20190222

摘要/Abstract

摘要： 目的：探讨多肽化合物urantide对动脉粥样硬化（AS）大鼠胸主动脉和血管平滑肌细胞（VSMC）中Ⅳ型胶原（Col Ⅳ）表达的影响，阐明其防治AS的作用机制。方法：180只Wistar大鼠随机分为正常对照组（n=30）和AS模型组（n=150），采用高脂饲料喂养和腹腔注射维生素D₃（VD₃）的方法建立大鼠AS模型。150只AS模型鼠随机分为AS组、氟伐他汀（Flu）组和urantide组（3、7和14 d组）（n=30）。免疫组织化学法检测大鼠胸主动脉壁内Col Ⅳ的表达水平，酶联免疫吸附试验（ELISA）法检测大鼠血清和尿液中羟脯胺酸（HYP）水平。体外培养的VSMC随机分为正常对照组、尾加压素（UⅡ）（10^-8 mol·L^-1）组、Flu组和urantide （10^-10~10^-6 mol·L^-1）组，ELISA法检测各组大鼠VSMC培养上清中Col Ⅳ水平。结果：各组大鼠胸主动脉内膜下不规则斑块内Col Ⅳ表达水平比较差异有统计学意义（F=35.09，P<0.01）。与正常对照组比较，AS组大鼠主动脉中Col Ⅳ表达水平明显升高（P<0.01）；urantide组Col Ⅳ阳性染色强度和范围较AS组均减少（P<0.01）。各组大鼠血清和尿液中HYP水平比较差异有统计学意义（F=24.38，P<0.01；F=26.72，P<0.01）。与正常对照组比较，AS组大鼠血清中HYP水平明显升高（P<0.01），尿液中HYP水平明显降低（P<0.01）；与AS组比较，urantide组大鼠血清中HYP水平均明显降低（P<0.01），尿液中HYP水平明显升高（P<0.01），接近或优于Flu组水平。各组大鼠VSMC培养上清中Col Ⅳ水平比较差异有统计学意义（F=31.04，P<0.01）。与正常对照组比较，UⅡ组大鼠VSMC培养上清中Col Ⅳ水平明显升高（P<0.01）；与UⅡ组比较，urantide组大鼠VSMC培养上清中Col Ⅳ水平明显降低（P<0.05或P<0.01）。结论：urantide可抑制AS大鼠胸主动脉和VSMC中Col Ⅳ的表达，缓解AS病变程度，为临床应用urantide治疗AS提供了实验依据。

关键词: 动脉粥样硬化, 尾加压素Ⅱ, urantide, 胸主动脉, 血管平滑肌细胞, Ⅳ型胶原

Abstract: Objective: To investigate the effects of urantide on the expressions of type Ⅳ collagen(Col Ⅳ) in thoracic aorta and vascular smooth muscle cells(VSMC) in the rats with atherosclerosis(AS), and to clarify its mechanism of prevention and treatment of AS.Methods: A total of 180 Wistar rats were randomly divided into normal control group (n=30) and AS model group (n=150).The rat models of AS were established by feeding on high-fat diet or intraperitoneally injecting vitamin D₃(VD₃).The AS model rats were randomly divided into AS group, fluvestation(Flu) group and urantide group (3, 7, and 14 d groups).The expression of Col Ⅳ in thoracic aorta wall of the rats was detected by immunohistochemistry.The levels of hydroxyproline (HYP) in serum and urine of the rats in various groups were measured by ELISA.The VSMC were randomly divided into normal control group, urotensin Ⅱ(UⅡ 10^-8mol·L^-1) group, Flu group and urantide (10^-10to 10^-6 mol·L^-1) groups.The levels of Col Ⅳ in VSMC of the rats in various groups were determined by ELISA.Results: There was a significant difference in the expression levels of Col Ⅳ in the irregular plaques of the thoracic aorta of the rats between various groups (F=35.09,P<0.01). The expression levels of Col Ⅳ in thoracic aorta of the rats in AS group were significantly increased compared with normal control group (P<0.01); the intensity and extent of Col Ⅳ positive staining in urantide group were lower than those in AS group (P<0.01).There were significant differences in the serum and urine HYP levels between various groups (F=24.38, P<0.01;F=26.72,P<0.01). Compared with normal control group, the serum HYP level of the rats in AS group were significantly increased (P<0.01), and the urine HYP level was significantly decreased (P<0.01).Compared with AS group, the serum HYP levels in urantide groups were significantly decreased (P<0.01) and the urine HYP levels were significantly increased (P<0.01), no less than the level in Flu group.The expression levels of Col Ⅳ in the culture supernatant of VSMC in the rats in various groups had significantly difference (F=31.04,P<0.01).The expression level of Col Ⅳ in the culture supernatant of VSMC of the rats in UⅡ group was significantly increased compared with normal control group (P<0.01); the expression levels of Col Ⅳ in the culture supernatant of VSMC of the rats in urantide groups were significantly decreased compared with UⅡ group(P<0.05 or P<0.01).Conclusion: Urantide can inhibit the expressions of Col Ⅳ in the thoracic aorta and VSMC of the AS rats and alleviate the degree of AS lesions, which provides the experimental evidence for the clinical application of urantide in the treatment of AS.

Key words: atherosclerosis, urotensinⅡ, urantide, vascular smooth muscle cells, type Ⅳ collagen

中图分类号:

R363

毕红东, 谢亚芹, 崔海鹏, 刘凯, 孙晓旭, 王途, 赵娟. 多肽化合物urantide对动脉粥样硬化大鼠胸主动脉和VSMC中Ⅳ型胶原表达的影响[J]. 吉林大学学报(医学版), 2019, 45(02): 342-346.

BI Hongdong, XIE Yaqin, CUI Haipeng, LIU Kai, SUN Xiaoxu, WANG Tu, ZHAO Juan. Effects of urantide on expressions of type Ⅳcollagenin thoracic aorta and VSMC of atherosclerotic rats[J]. Journal of Jilin University(Medicine Edition), 2019, 45(02): 342-346.

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