晚期糖基化终末产物诱导大鼠主动脉平滑肌细胞转分化的研究  

The study of the induction of advanced glycosylation end products on transdifferentiation of rat aortic smooth muscle cells

作　　者：刘超[1] 谭若芸[1] 殷长军[1] 顾民[1] 

机构地区：[1]南京医科大学第一附属医院泌尿外科,210029

出　　处：《中华实验外科杂志》2013年第5期905-907,共3页Chinese Journal of Experimental Surgery

基　　金：基金项目：江苏省卫生厅面上项目（H2009907）;江苏省“六大人才高峰人才”项目（2010WSN-56）;江苏省“科教兴卫”工程医学重点人才培养资助项目（RC2011055）;江苏省“333高层次人才工程”培养基金资助项目（2011）;中华医学会临床医学科研专项基金资助项目（09010310186）;江苏省自然科学基金资助项目（BK2008482）;国家自然科学基金青年基金资助项目（81100532）

摘　　要：目的探讨晚期糖基化终末产物（AGEs）参与肾移植术后动脉粥样硬化形成的机制。方法原代培养SD大鼠主动脉平滑肌细胞；用AGEs或牛血清白蛋白200mg／L作用该细胞不同时程（12h～12d），采用Western blot法和间接免疫荧光法检测eL．平滑肌肌动蛋白（α-SMA）、特异性核转录斟子（RUNX2）和骨桥蛋白（OPN）的表达。结果第2～5代VSMCs中α-SMA高度表达；在AGEs作用后α-SMA表达量及阳性细胞数进行性减少；RUNX2及OPN表达进行性升高。Western blot检测α-SMA依次为：1．1170±0．1080、1．1820±0．1313、1．1060±0．0738、0．8360±0．0329、0．7430±0．0504、0．5790±0．0718、0．4600±0．1007、0．2230±0．0658、0．1810±0．0427；OPN依次为：0．0320±0．0019、0．0330±0．0010、0．0380±0．0054、0．0470±0．0077、0．0380±0．0232、0．1520±0．0460、0．2220±0．0536、0．4040±0．0290、0．6730±0．0495；RUNX2依次为：0．3640±0．0477、0．3690±0．0133、0．4840±0．0462、0．5290±0．0452、0．8850±0．0788、1．1500±0．1001、1．3460±0．1216，各组间差异有统计学意义（P〈0．05）。结论AGEs可能通过上调RUNX2表达诱导VSMCs向成骨细胞转分化，进而促进肾移植术后动脉粥样硬化的进展。Objective To investigate the mechanism of advanced glycosylation end products （AGEs） involved in the progression of artheroselerosis after kidney transplantion. Methods The fifth Sprague-Dawley （SD） rat aortic vascular smooth muscle cells （VSMCs） was an in vitro culture system. The VSMCs were incubated with 200 mg/L AGEs or bovine serum albumin （BSA） for 12 h-12 d, then the cells were collected and detected the expressions of alpha-smooth muscle actin （α-SMA） , human runt-related transcription factor 2 （RUNX2） and osteopontin （OPN） by Western blotting and indirect immunofluorescence staining assays. Results The abundance of α-SMA was very high in the second to fifth SD rat aortic VSMCs. Compared with normal control or the ceils treated with BSA, AGEs can reduce α-SMA protein expression and the percent of α-SMA positive cells in rat aortic VSMCs in a time-dependent manner. Meanwhile, the expression of RUNX2 protein was rapidly induced after AGEs treatment for 12 h, and followed up, the expression of OPN was also increased in rat aortic VSMCs cells. The expression levels of α-SMA detected by Western blot were as follow： 1. 1170 ±0. 1080, 1. 1820 ±0. 1313, 1. 1060 ±0. 0738, 0. 8360 ±0. 0329, 0. 7430 ±0. 0504, 0. 5790 ± 0. 0718, 0. 4600 ± 0. 1007, 0. 2230 ± 0. 0658, 0. 1810 ± 0. 0427 ; OPN- 0. 0320 ±0 . 0019, 0. 0330 ± 0. 0010, 0. 0380 ± 0. 0054, 0. 0470 ± 0. 0077, 0. 0380 ± 0. 0232, 0. 1520 ±0. 0460, 0. 2220 ±0. 0536, 0. 4040 ±0. 0290, 0. 6730 ±0. 0495; RUNX2： 0. 3640 ± 0. 0477, 0. 3690 ± 0. 0133, 0. 4840 ± 0. 0462, 0. 5290 ± 0. 0452, 0. 8850 ± 0. 0788, 1. 1500 ± 0. 1001, 1. 3460 ± 0. 1216 （ all P 〈 0. 05 ）. Conclusion AGEs maybe act as a key role in the procession of artherosclerosis after kidney transpiantion through inducing transdifferentiation of VSMCs to osteoblast-like cells by upregulation of RUNX2.

关 键 词：肾移植术 动脉粥样硬化 晚期糖基化终末产物 血管平滑肌细胞 成骨细胞 转分化 

分 类 号：R5[医药卫生—内科学]