中华肾脏病杂志
中華腎髒病雜誌
중화신장병잡지
2015年
5期
351-358
,共8页
汤日宁%朱冬冬%韩雨晨%伍敏%吕林莉%马坤岭%刘必成
湯日寧%硃鼕鼕%韓雨晨%伍敏%呂林莉%馬坤嶺%劉必成
탕일저%주동동%한우신%오민%려림리%마곤령%류필성
纤维化%糖尿病%细胞转分化%厄贝沙坦
纖維化%糖尿病%細胞轉分化%阨貝沙坦
섬유화%당뇨병%세포전분화%액패사탄
Fibrosis%Diabetes%Cell transdifferentiation%Irbesartan
目的 探讨厄贝沙坦对糖尿病大鼠心肌内皮细胞转分化(EndMT)的影响.方法 体内实验:采用腹腔注射链脲佐菌素(STZ)的方法制备自发性高血压症大鼠(SHR)糖尿病模型(n=8).模型组大鼠分为糖尿病组和厄贝沙坦(Isb)治疗组;Wistar-Kyoto (WKY)大鼠作为对照组.体外实验:人主动脉内皮细胞(HAEC)被分为对照组(NG,5.5 mmol/L);高糖组(HG,30 mmol/L糖);厄贝沙坦组(HG+Irb,HG+Irb 1 μmol/L)3组.光镜下观察各组大鼠心肌组织病理学改变;透射电镜观察心肌微血管内皮细胞病理改变.激光扫描共聚焦显微镜(LSCM)免疫荧光染色方法观察心肌和高糖刺激的人主动脉内皮细胞CD31和成纤维细胞特异蛋白1(FSP1)的表达.放射免疫法检测HAEC上清液中血管紧张素Ⅱ的浓度.Western印迹法检测FSP1、α平滑肌肌动蛋白(α-SMA)的表达.结果 与WKY组比较,糖尿病组大鼠出现明显的心肌纤维化.透射电镜结果显示:糖尿病组大鼠心肌微血管内皮指状突起增多.LSCM结果显示心肌CD31和FSP1共表达.体外实验结果显示:与对照组相比,高糖组HAEC细胞上清液中血管紧张素Ⅱ的浓度增加(P＜0.05),LSCM可见CD31和FSP1表达重叠,部分细胞获得纺锤样改变并失去CD31染色.高糖组FSP1和α-SMA蛋白水平的表达明显增加(均P＜ 0.05),厄贝沙坦组细胞上述改变减轻(P＜0.05).结论 厄贝沙坦可通过抑制心肌EndMT,减轻糖尿病心肌纤维化.
目的 探討阨貝沙坦對糖尿病大鼠心肌內皮細胞轉分化(EndMT)的影響.方法 體內實驗:採用腹腔註射鏈脲佐菌素(STZ)的方法製備自髮性高血壓癥大鼠(SHR)糖尿病模型(n=8).模型組大鼠分為糖尿病組和阨貝沙坦(Isb)治療組;Wistar-Kyoto (WKY)大鼠作為對照組.體外實驗:人主動脈內皮細胞(HAEC)被分為對照組(NG,5.5 mmol/L);高糖組(HG,30 mmol/L糖);阨貝沙坦組(HG+Irb,HG+Irb 1 μmol/L)3組.光鏡下觀察各組大鼠心肌組織病理學改變;透射電鏡觀察心肌微血管內皮細胞病理改變.激光掃描共聚焦顯微鏡(LSCM)免疫熒光染色方法觀察心肌和高糖刺激的人主動脈內皮細胞CD31和成纖維細胞特異蛋白1(FSP1)的錶達.放射免疫法檢測HAEC上清液中血管緊張素Ⅱ的濃度.Western印跡法檢測FSP1、α平滑肌肌動蛋白(α-SMA)的錶達.結果 與WKY組比較,糖尿病組大鼠齣現明顯的心肌纖維化.透射電鏡結果顯示:糖尿病組大鼠心肌微血管內皮指狀突起增多.LSCM結果顯示心肌CD31和FSP1共錶達.體外實驗結果顯示:與對照組相比,高糖組HAEC細胞上清液中血管緊張素Ⅱ的濃度增加(P＜0.05),LSCM可見CD31和FSP1錶達重疊,部分細胞穫得紡錘樣改變併失去CD31染色.高糖組FSP1和α-SMA蛋白水平的錶達明顯增加(均P＜ 0.05),阨貝沙坦組細胞上述改變減輕(P＜0.05).結論 阨貝沙坦可通過抑製心肌EndMT,減輕糖尿病心肌纖維化.
목적 탐토액패사탄대당뇨병대서심기내피세포전분화(EndMT)적영향.방법 체내실험:채용복강주사련뇨좌균소(STZ)적방법제비자발성고혈압증대서(SHR)당뇨병모형(n=8).모형조대서분위당뇨병조화액패사탄(Isb)치료조;Wistar-Kyoto (WKY)대서작위대조조.체외실험:인주동맥내피세포(HAEC)피분위대조조(NG,5.5 mmol/L);고당조(HG,30 mmol/L당);액패사탄조(HG+Irb,HG+Irb 1 μmol/L)3조.광경하관찰각조대서심기조직병이학개변;투사전경관찰심기미혈관내피세포병리개변.격광소묘공취초현미경(LSCM)면역형광염색방법관찰심기화고당자격적인주동맥내피세포CD31화성섬유세포특이단백1(FSP1)적표체.방사면역법검측HAEC상청액중혈관긴장소Ⅱ적농도.Western인적법검측FSP1、α평활기기동단백(α-SMA)적표체.결과 여WKY조비교,당뇨병조대서출현명현적심기섬유화.투사전경결과현시:당뇨병조대서심기미혈관내피지상돌기증다.LSCM결과현시심기CD31화FSP1공표체.체외실험결과현시:여대조조상비,고당조HAEC세포상청액중혈관긴장소Ⅱ적농도증가(P＜0.05),LSCM가견CD31화FSP1표체중첩,부분세포획득방추양개변병실거CD31염색.고당조FSP1화α-SMA단백수평적표체명현증가(균P＜ 0.05),액패사탄조세포상술개변감경(P＜0.05).결론 액패사탄가통과억제심기EndMT,감경당뇨병심기섬유화.
Objective To explore the effect of irbesartan on cardiac endothelial-mesenchymal transition (EndMT) in diabetic rats.Methods The model of diabetic rat was induced by intraperitoneal injection with streptozotocin (STZ,35 mg/kg) in spontaneous hypertensive rats (SHR).Diabetic rats were divided into diabetic group and the Irbesartan treated group.The pathological changes were investigated by fluorescence microscope and electron microscope.The EndMT was studied in human aortic endothelial cells (HAEC) exposure to high glucose.The concentration of angiotensin Ⅱ in the supernatant was detected by radioimmunoassay.Immunofluorescence staining was performed to detect the co-localization of CD31 and FSP1.Results The significant myocardial fibrosis was presented in the diabetic group.Endothelial protrusions were prominent feature in myocardial microvascular of diabetic rat compared with the control group rats.Double staining of HAEC showed co-localization of CD31 and FSP1,which was decreased by the treatment of Irbesartan (P ＜ 0.05).When HAEC was exposed to high glucose,it showed some cells acquired spindle-shaped morphology and lost CD31 staining,and FSP1 and α-SMA protein expression levels were markedly upregulated,which attenuated by the treatment of Irbesartan.Conclusion Irbesartan might prevent diabetes from myocardial fibrosis via inhibition of EndMT in diabetic rats.
