西红花酸对晚期糖基化终产物诱导牛血管内皮细胞E-选择素表达的抑制作用

中国临床药理学与治疗学 ›› 2010, Vol. 15 ›› Issue (7): 764-769.

西红花酸对晚期糖基化终产物诱导牛血管内皮细胞E-选择素表达的抑制作用

向敏¹, 周成华², 钱之玉³

¹苏州卫生职业技术学院生物技术中心,苏州 215009,江苏;
²徐州医学院药理学教研室, 徐州 221004,江苏;
³中国药科大学药学院药理学教研室,南京 210009,江苏

收稿日期:2010-06-02 修回日期:2010-07-12 出版日期:2010-07-26 发布日期:2020-09-15
通讯作者: 钱之玉,男,教授,博导,研究方向:生化药理。Tel: 0512-62690089 E-mail: xiangmin99@126.com
作者简介:向敏,女,博士,副教授,研究方向:糖尿病血管病变。

Inhibitory action of crocetin on the expression of E-selectin in bovine endothelial cells induced by advanced glycation end products

XIANG Min¹, ZHOU Cheng-hua², QIAN Zhi-yu³

¹Biotechnology Center, Suzhou Health College, Suzhou 215009, Jiangsu, China;
²Department of Pharmacology, Xuzhou Medical College, Xuzhou 221004, Jiangsu, China;
³Department of Pharmacology, China Pharmaceutical University, Nanjing 210009, Jiangsu, China

Received:2010-06-02 Revised:2010-07-12 Online:2010-07-26 Published:2020-09-15

摘要/Abstract

摘要： 目的: 研究西红花酸(crocetin)对晚期糖基化终产物(advanced glycation end products,AGEs)诱导牛主动脉血管内皮细胞(bovine Endothelial cells, BECs)E-选择素(E-selectin)表达的抑制作用。方法: 分离纯化新生牛全血的中性粒细胞与单核细胞,用虎红染色法测定西红花酸对牛单核细胞/中性粒细胞与BECs黏附的影响,Cell-based ELISA法和sABC免疫细胞化学法测定E-选择素蛋白表达变化。结果: AGEs(100 μg/mL)能促进中性粒细胞和单核细胞对BECs的黏附(P<0.01 vs control),用西红花酸(1,0.1, 0.01 μmol/L)预孵内皮细胞 12 h 后,再用AGEs作用 24 h, 中性粒细胞和单核细胞对BECs的黏附较AGEs模型组降低,且呈剂量依赖性关系(P<0.05或0.01)。Cell-based ELISA测定结果显示,AGEs作用 4 h 内,BECs中的E-selectin表达水平上升,之后E-selectin表达下降,8 h 时,恢复接近正常水平。而西红花酸 (1, 0.1 μmol/L)能使E-selectin表达下降。sABC免疫化学结果也证实西红花酸对AGEs诱导BECs中E-selectin表达有抑制作用。结论: 西红花酸可通过抑制黏附分子E-selcetin蛋白表达,从而抑制中性粒细胞和单核细胞对内皮细胞的黏附作用,这可能是西红花酸减轻糖尿病血管病变的作用机制之一。

关键词: 西红花酸, 黏附, E-选择素, 中性粒细胞, 单核细胞, 内皮细胞

Abstract: AIM: To study the inhibitory effect of crocetin on the expression of E-selectin in bovine endothelial cells (BECs) induced by advanced glycation end products (AGEs).METHODS: Neutrophils and monocytes were separated and purified from the whole blood of new-born calf. The adhesion of neutrophils/ monocytes to endothelial cells was assayed by Fuhong staining method. Cell-based ELISA and sABC immunocytochemistry were adopted to analyze the expression of E-selectin.RESULTS: AGEs(100 μg/mL)could increase the adhesion of neutrophils/monocytes to endothelial cells(P<0.01 vs control). However, after pre-incubation the BECs with crocetin(1,0.1,0.01 μmol/L) for 12 h before exposure to AGEs (100 mg/mL), the above adhesion rate decreased significantly in a dose-dependent manner( P<0.05 or 0.01 vs AGEs group). Cell-based ELISA results demonstrated that the expression of E-selectin elevated in BECs treated by AGEs within 4 h, then decreased and regained to the normal level at 8 h, while the protein expression of E-selectin was suppressed by crocetin. sABC immunocytochemistry also confirmed that crocetin could decrease the level of E-selectin.CONCLUSION: These results demonstrate that crocetin could inhibit E-selectin over-expression in BECs exposed to AGEs and then down-regulate the adhesion of neutrophils/monocytes to endothelial cells, which is one of the possible mechanisms for crocetin to attenuate diabetic vascular complications.

Key words: Crocetin, Adhesion, E-selectin , Neutrophil, Monocyte, Endothelial cell

中图分类号:

R965.2

向敏, 周成华, 钱之玉. 西红花酸对晚期糖基化终产物诱导牛血管内皮细胞E-选择素表达的抑制作用[J]. 中国临床药理学与治疗学, 2010, 15(7): 764-769.

XIANG Min, ZHOU Cheng-hua, QIAN Zhi-yu. Inhibitory action of crocetin on the expression of E-selectin in bovine endothelial cells induced by advanced glycation end products[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2010, 15(7): 764-769.

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