[1]廖云鹏,何 芳,江 芬,等.环氧化酶-2与骨形态发生蛋白9诱导血管平滑肌钙化的关系研究[J].中国药理学通报,2018,(08):1066-1072.[doi:10.3969/j.issn.1001-1978.2018.08.008]
 LIAO Yun-peng,HE Fang,JIANG Fen,et al.Study on the relationship between BMP9-induced calcification and COX-2 in VSMCs[J].Chinese Pharmacological Bulletin,2018,(08):1066-1072.[doi:10.3969/j.issn.1001-1978.2018.08.008]
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环氧化酶-2与骨形态发生蛋白9诱导血管平滑肌钙化的关系研究()
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《中国药理学通报》[ISSN:/CN:]

卷:
期数:
2018年08期
页码:
1066-1072
栏目:
论著
出版日期:
2018-07-26

文章信息/Info

Title:
Study on the relationship between BMP9-induced calcification and COX-2 in VSMCs
文章编号:
1001-1978(2018)08-1066-07
作者:
廖云鹏12何 芳13江 芬124王 涵12朱茄慧12马 妍12胡 莹12李福书12李 沁12周 娅12何百成12
1.重庆市生物化学与分子药理学重点实验室; 2.重庆医科大学药理学教研室; 3.重庆医科大学附属第一医院肾内科,重庆 400016; 4.湖北省蕲春县人口和计划生育服务中心,湖北 蕲春 436300
Author(s):
LIAO Yun-peng12 HE Fang13 JIANG Fen124 WANG Han12 ZHU Jia-hui12MA Yan12 HU Ying12 LI Fu-shu12 LI Qin12 ZHOU Ya12 HE Bai-chen
1.Chongqing Key Lab of Biochemistry and Molecular Pharmacology, Chongqing 400016, China; 2.Dept of Pharmacology,Pharmacy School of Chongqing Medical University, Chongqing 400016, China; 3.Dept of Nephrology,the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China; 4.Population and Birth Controlling Service Center of Qichun County, Qichun Hubei 436300, China
关键词:
主动脉 血管钙化 高磷酸盐 血管平滑肌细胞 骨形态发生蛋白9 环氧化酶-2
Keywords:
aorta vascular calcification high phosphate vascular smooth muscle cells BMP9 COX-2
分类号:
R-332; R322.121; R322.74; R692.022; R977.3; R977.6
DOI:
10.3969/j.issn.1001-1978.2018.08.008
文献标志码:
A
摘要:
目的 研究高磷对骨形态发生蛋白9(bone morphogenetic protein 9, BMP9)的影响,BMP9对血管平滑肌细胞(vascular smooth muscle cells,VSMCs)钙化的影响与环氧酶-2(cyclooxygenase-2, COX-2)的关系。方法 利用免疫荧光、茜素红染色和Western blot检测高磷水平对血管平滑肌钙化的诱导作用; 利用茜素红染色、硝酸银染色、real-time PCR和Western blot检测BMP9诱导血管平滑肌钙化的作用; 采用免疫荧光、茜素红染色、real-time PCR、Western blot检测COX-2与BMP9诱导血管平滑肌钙化的关系。结果 与对照组相比,高磷明显促进骨桥蛋白(osteopontin, OPN)和骨钙素(osteocalin, OCN)表达,并诱导VSMCs出现明显钙盐沉积; 高磷呈浓度依赖性方式诱导BMP9表达,增加Runx2、Dlx-5、ALP等骨向调节因子的mRNA表达水平,但降低SM22α的mRNA。BMP9过表达明显增加VSMCs中OPN和OCN的表达水平,同时降低α-平滑肌肌动蛋白(α-smooth muscle actin, α-SMA)的表达水平; BMP9过表达明显诱导钙盐沉积,但ALK2突变后则减弱BMP9的这种作用; BMP9过表达能诱导血管平滑肌钙化,14 d时作用最明显。高磷和BMP9过表达都能在VMSCs中明显诱导COX-2表达,过表达COX-2增强BMP9在VSMCs中对OPN和OCN表达的促进作用,但加强BMP9对α-SMA表达的抑制作用; COX-2抑制剂减弱BMP9对OPN和OCN表达的促进作用, 以及BMP9对α-SMA表达的抑制作用; COX-2抑制剂同时还降低BMP9在VSMCs中促进钙盐沉积的作用。 结论 高磷诱导VSMCs钙化可能与促进BMP9和COX-2表达有关,并且COX-2参与介导BMP9诱导的VMSCs钙化。
Abstract:
Aim To investigate the relationship between phosphate and BMP9 in VSMCs, and the effect of BMP9 and/or COX-2 on the calcification in VSMCs.Methods We used immunofluorescent, Alizarin Red S stain and Western blot to analyze the effect of high phosphate on calcification in VSMCs.Alizarin Red S staining, Von Kossa staining, real-time PCR and Western blot were employed to analyze the effect of BMP9 on calcification in VSMCs.Western blot was used to determine the effect of phosphate and BMP9 on the expression of COX-2 in VSMCs.Western blot and Alizarin Red S staining were employed to determine whether COX-2 was involved in BMP9-induced calcification in VSMCs.Results High phosphate induced the expression of OPN and OCN, so did calcification apparently.Phosphate increased the expression of BMP9 and over-expression of BMP9 induced the expression of Runx2, Dlx-5 and ALP in VSMCs, but decreased the level of SM22α.BMP9 could also increase the mineralization in VSMCs, but it was down-regulated by the dominant negative mutation of ALK2.Phosphate and BMP9 both induced the expression of COX-2 in VSMCs.Over-expression of COX-2 increased the level of BMP9-induced OPN and OCN, but enhanced the effect of BMP9 on decreasing the level of α-SMA.The COX-2 specific inhibitor showed the reverse effects on OPN, OCN and α-SMA.Meanwhile, COX-2 specific inhibitor also decreased the BMP9-induced mineralization in VSMCs.Conclusions Phosphate can up-regulate BMP9 and COX-2 when calcification is induced in VSMCs, and COX-2 may mediate the BMP9-induced calcification in VSMCs.

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期:2018-04-17,修回日期:2018-05-20
基金项目:国家自然科学基金资助项目(No 81572226)
作者简介:廖云鹏(1993-),男,硕士生,研究方向:分子药理学,E-mail:liaoyunpeng93@sina.com;
何百成(1972-),男,博士,教授,研究方向:分子药理学和干细胞生物学,通讯作者,E-mail:hebaicheng99@yahoo.com
更新日期/Last Update: 2018-07-26