中华劳动卫生职业病杂志
中華勞動衛生職業病雜誌
중화노동위생직업병잡지
CHINESE JOURNAL OF INDUSTRIAL HYGIENE AND OCCUPATIONAL DISEASES
2014年
11期
801-805
,共5页
李世峰%杜世璞%薛新新%徐丁洁%徐洪%孙月%邓海静%杨奕%魏中秋
李世峰%杜世璞%薛新新%徐丁潔%徐洪%孫月%鄧海靜%楊奕%魏中鞦
리세봉%두세박%설신신%서정길%서홍%손월%산해정%양혁%위중추
N-乙酰基-丝氨酰-天门冬氨酰-赖氨酰-脯氨酸%血管紧张素Ⅱ%成纤维细胞%肺纤维化
N-乙酰基-絲氨酰-天門鼕氨酰-賴氨酰-脯氨痠%血管緊張素Ⅱ%成纖維細胞%肺纖維化
N-을선기-사안선-천문동안선-뢰안선-포안산%혈관긴장소Ⅱ%성섬유세포%폐섬유화
N-acetyl-seryl-aspartyl-lysyl-proline%Angiotensin Ⅱ%Fibroblast%Lung fibrosis
目的 研究N-乙酰基-丝氨酰-天门冬氨酰-赖氨酰-脯氨酸(Ac-SDKP)对血管紧张素(Ang)Ⅱ诱导的人胚肺MRC-5成纤维细胞向肌成纤维细胞分化的调节作用.方法 实验分为2部分:(1)采用不同浓度AngⅡ诱导48 h,采用100 nmol/L AngⅡ诱导不同时间点,免疫印迹法检测Ⅰ型胶原和α-平滑肌肌动蛋白(α-SMA)的表达;(2)实验分组为对照组、AngⅡ诱导组、Ac-SDKP干预组、cAMP直接激活的交换蛋白(Epac)蛋白特异性激活剂(8-Me-cAMP)干预组,免疫细胞化学染色法观察α-SMA的表达,免疫印迹法检测Ⅰ型胶原、α-SMA、血清反应因子(SRF)及其转录辅因子肌相关转录因子(MRTF)-A、Epac 1、2的表达.结果 AngⅡ能够明显诱导MRC-5细胞Ⅰ型胶原和α-SMA的表达,并具有一定的剂量和时间依赖效应.免疫细胞化学染色可见AngⅡ诱导组细胞质内出现明显的α-SMA阳性显色,同时诱导组SRF、MRTF-A、α-SMA和Ⅰ型胶原蛋白表达上调,分别是对照组的3.4、3.5、3.3、6.8倍,Epac1蛋白表达下调,差异均有统计学意义(P＜0.05);与AngⅡ诱导组比较,8-Me-cAMP干预组和Ac-SDKP干预组胞质内α-SMA阳性显色减弱,同时SRF、MRTF-A、α-SMA和Ⅰ型胶原蛋白表达下调,Epac1蛋白表达上调,差异均有统计学意义(P＜0.05).结论 Ac-SDKP能够通过上调Epac1蛋白抑制AngⅡ诱导人胚肺MRC-5成纤维细胞向肌成纤维细胞分化.
目的 研究N-乙酰基-絲氨酰-天門鼕氨酰-賴氨酰-脯氨痠(Ac-SDKP)對血管緊張素(Ang)Ⅱ誘導的人胚肺MRC-5成纖維細胞嚮肌成纖維細胞分化的調節作用.方法 實驗分為2部分:(1)採用不同濃度AngⅡ誘導48 h,採用100 nmol/L AngⅡ誘導不同時間點,免疫印跡法檢測Ⅰ型膠原和α-平滑肌肌動蛋白(α-SMA)的錶達;(2)實驗分組為對照組、AngⅡ誘導組、Ac-SDKP榦預組、cAMP直接激活的交換蛋白(Epac)蛋白特異性激活劑(8-Me-cAMP)榦預組,免疫細胞化學染色法觀察α-SMA的錶達,免疫印跡法檢測Ⅰ型膠原、α-SMA、血清反應因子(SRF)及其轉錄輔因子肌相關轉錄因子(MRTF)-A、Epac 1、2的錶達.結果 AngⅡ能夠明顯誘導MRC-5細胞Ⅰ型膠原和α-SMA的錶達,併具有一定的劑量和時間依賴效應.免疫細胞化學染色可見AngⅡ誘導組細胞質內齣現明顯的α-SMA暘性顯色,同時誘導組SRF、MRTF-A、α-SMA和Ⅰ型膠原蛋白錶達上調,分彆是對照組的3.4、3.5、3.3、6.8倍,Epac1蛋白錶達下調,差異均有統計學意義(P＜0.05);與AngⅡ誘導組比較,8-Me-cAMP榦預組和Ac-SDKP榦預組胞質內α-SMA暘性顯色減弱,同時SRF、MRTF-A、α-SMA和Ⅰ型膠原蛋白錶達下調,Epac1蛋白錶達上調,差異均有統計學意義(P＜0.05).結論 Ac-SDKP能夠通過上調Epac1蛋白抑製AngⅡ誘導人胚肺MRC-5成纖維細胞嚮肌成纖維細胞分化.
목적 연구N-을선기-사안선-천문동안선-뢰안선-포안산(Ac-SDKP)대혈관긴장소(Ang)Ⅱ유도적인배폐MRC-5성섬유세포향기성섬유세포분화적조절작용.방법 실험분위2부분:(1)채용불동농도AngⅡ유도48 h,채용100 nmol/L AngⅡ유도불동시간점,면역인적법검측Ⅰ형효원화α-평활기기동단백(α-SMA)적표체;(2)실험분조위대조조、AngⅡ유도조、Ac-SDKP간예조、cAMP직접격활적교환단백(Epac)단백특이성격활제(8-Me-cAMP)간예조,면역세포화학염색법관찰α-SMA적표체,면역인적법검측Ⅰ형효원、α-SMA、혈청반응인자(SRF)급기전록보인자기상관전록인자(MRTF)-A、Epac 1、2적표체.결과 AngⅡ능구명현유도MRC-5세포Ⅰ형효원화α-SMA적표체,병구유일정적제량화시간의뢰효응.면역세포화학염색가견AngⅡ유도조세포질내출현명현적α-SMA양성현색,동시유도조SRF、MRTF-A、α-SMA화Ⅰ형효원단백표체상조,분별시대조조적3.4、3.5、3.3、6.8배,Epac1단백표체하조,차이균유통계학의의(P＜0.05);여AngⅡ유도조비교,8-Me-cAMP간예조화Ac-SDKP간예조포질내α-SMA양성현색감약,동시SRF、MRTF-A、α-SMA화Ⅰ형효원단백표체하조,Epac1단백표체상조,차이균유통계학의의(P＜0.05).결론 Ac-SDKP능구통과상조Epac1단백억제AngⅡ유도인배폐MRC-5성섬유세포향기성섬유세포분화.
Objective To explore the inhibition effect of N-acetyl-seryl-aspartyl-lysyl-proline (AcSDKP) on myofibroblast differentiation of MRC-5 human fetal lung fibroblasts induced by angiotensin (Ang)Ⅱ.Methods The study was divided into 2 step:(1) MRC-5 human fetal lung fibroblasts was induced for 48 h at different dose of Ang Ⅱ and at different time point by 100 nmol/L Ang Ⅱ.Then the expression of collagen type Ⅰ and α-smooth muscle actin (α-SMA) were mesaured by western blot.(2) MRC-5 human fetal lung fibroblasts were divided into 4 group:(1) control,(2) Ang Ⅱ,(3) Ang Ⅱ +Ac-SDKP,(4) Ang Ⅱ +8-Me-cAMP (a specific activator of Epac).The α-SMA expression was observed by immnocytochemical stain.The protein expression of collagen type Ⅰ,α-SMA,serum response factor (SRF),myocardin-related transcription factor (MRTF)-A,exchange protein directly activated by cAMP (Epac) 1,2 were measured by Westen blot.Results Myofibroblast differentiation could be induced by Ang Ⅱ from MRC-5 cells with a dose-and time-dependent manner.The up-regulation of SRF and MRTF-A were observed in MRC-5 cells induced by Ang Ⅱ and accompanied with collagen Ⅰ and α-SMA increased.Pre-treatment with 8-Me-cAMP or Ac-SDKP could attenuated all this changes induced by Ang Ⅱ,and promoted the expression of Epac1.Conclusion Ac-SDKP can inhibit the myofibroblast differentiation of MRC-5 cells induced by Ang Ⅱ via Epac 1 activating.
