中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
5期
905-907
,共3页
刘超%谭若芸%殷长军%顾民
劉超%譚若蕓%慇長軍%顧民
류초%담약예%은장군%고민
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
腎移植術%動脈粥樣硬化%晚期糖基化終末產物%血管平滑肌細胞%成骨細胞%轉分化
신이식술%동맥죽양경화%만기당기화종말산물%혈관평활기세포%성골세포%전분화
Kidney transplantion%Artherosclerosis%Advanced glycosylation end products%Vascular smooth muscle cells%Osteoblast cells%Transdifferentiation
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200 mg/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α-平滑肌肌动蛋白(α-SMA)、特异性核转录因子(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±00477、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.
目的 探討晚期糖基化終末產物(AGEs)參與腎移植術後動脈粥樣硬化形成的機製.方法 原代培養SD大鼠主動脈平滑肌細胞；用AGEs或牛血清白蛋白200 mg/L作用該細胞不同時程(12h～12d),採用Western blot法和間接免疫熒光法檢測α-平滑肌肌動蛋白(α-SMA)、特異性覈轉錄因子(RUNX2)和骨橋蛋白(OPN)的錶達.結果 第2～5代VSMCs中α-SMA高度錶達；在AGEs作用後α-SMA錶達量及暘性細胞數進行性減少;RUNX2及OPN錶達進行性升高.Western blot檢測α-SMA依次為: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依次為: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依次為:0.3640±00477、0.3690±0.0133、0.4840±0.0462、0.5290±0.0452、0.8850±0.0788、1.1500±0.1001、1.3460±0.1216,各組間差異有統計學意義(P＜0.05).結論 AGEs可能通過上調RUNX2錶達誘導VSMCs嚮成骨細胞轉分化,進而促進腎移植術後動脈粥樣硬化的進展.
목적 탐토만기당기화종말산물(AGEs)삼여신이식술후동맥죽양경화형성적궤제.방법 원대배양SD대서주동맥평활기세포；용AGEs혹우혈청백단백200 mg/L작용해세포불동시정(12h～12d),채용Western blot법화간접면역형광법검측α-평활기기동단백(α-SMA)、특이성핵전록인자(RUNX2)화골교단백(OPN)적표체.결과 제2～5대VSMCs중α-SMA고도표체；재AGEs작용후α-SMA표체량급양성세포수진행성감소;RUNX2급OPN표체진행성승고.Western blot검측α-SMA의차위: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;OPN의차위: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; RUNX2의차위:0.3640±00477、0.3690±0.0133、0.4840±0.0462、0.5290±0.0452、0.8850±0.0788、1.1500±0.1001、1.3460±0.1216,각조간차이유통계학의의(P＜0.05).결론 AGEs가능통과상조RUNX2표체유도VSMCs향성골세포전분화,진이촉진신이식술후동맥죽양경화적진전.
Objective To investigate the mechanism of advanced glycosylation end products (AGEs) involved in the progression of artherosclerosis 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.Coinpared with normal control or the cells 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 transplantion through indueing transdifferentiation of VSMCs to osteoblast-like cells by upregulation of RUNX2.
