中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2011年
26期
1847-1851
,共5页
尚峰%何志旭%汪浩文%崔冬冰%杨燕
尚峰%何誌旭%汪浩文%崔鼕冰%楊燕
상봉%하지욱%왕호문%최동빙%양연
高血压,肺性%间质干细胞移植%血管内皮生长因子类%基因
高血壓,肺性%間質榦細胞移植%血管內皮生長因子類%基因
고혈압,폐성%간질간세포이식%혈관내피생장인자류%기인
Hypertension,pulmonary%Mesenchymal stem cell transplantation%Vascular endothelial growth factors%Genes
目的 探讨血管内皮生长因子(VEGF)基因转染的骨髓间充质干细胞(MMSC)移植治疗肺动脉高压(PH)的疗效和机制.方法 体外分离培养SD大鼠MMSC,将含有增强型绿色荧光蛋白的VEGF质粒转进MMSC.将SD大鼠随机分为4组:正常对照组、PH模型组;MMSC移植组,转基因移植组.一次性项背部皮下注射野百合碱(50 mg/kg),复制PH模型,观察21 d后,MMSC移植组及转基因移植组分别给予浓度为5×106个细胞/ml和5×105个细胞/ml的MMSC悬液1 ml静脉注射,模型组及正常组均给予等量L-DMEM液注射.移植后30 d,观察4组大鼠一般情况、生存率、右心室收缩压(RVSP)、右心室肥大指数(RVH)、血气及肺小动脉的微观结构变化.结果 移植后30 d,MMSC移植组及转基因移植组大鼠的RVSP分别为(30.2 ±2.1)和(29.2±1.1)mm Hg(1 mm Hg=0.133 kPa),RVH分别为(37.9 ±3.2)%和(27.2±3.4)%,肺小动脉中膜厚度(MT)分别为(21.3±3.4)和(14.3±2.8)μm,肺小动脉管腔横截面积与血管总横截面积的比值(V/T)分别为(39.3 ±4.3)%和(43.0 ±1.5)%.上述指标与PH模型组相比差异均有统计学意义(P<0.01);其中MMSC移植组与转基因移植组间RVH、MT及V/T的差异有统计学意义(P<0.05).另外,MMSC移植组与转基因移植组动脉血气各指标较PH模型组均明显改善(P<0.05).同时观察到肺内损伤血管修复和血管新牛现象.结论 转染有VEGF165基因的MMSC移植可有效减轻并逆转野百合碱诱导的PH进程,此作用较单纯移植MMSC的治疗效果更加明显.其作用机制可能与血管修复及血管新生有关.
目的 探討血管內皮生長因子(VEGF)基因轉染的骨髓間充質榦細胞(MMSC)移植治療肺動脈高壓(PH)的療效和機製.方法 體外分離培養SD大鼠MMSC,將含有增彊型綠色熒光蛋白的VEGF質粒轉進MMSC.將SD大鼠隨機分為4組:正常對照組、PH模型組;MMSC移植組,轉基因移植組.一次性項揹部皮下註射野百閤堿(50 mg/kg),複製PH模型,觀察21 d後,MMSC移植組及轉基因移植組分彆給予濃度為5×106箇細胞/ml和5×105箇細胞/ml的MMSC懸液1 ml靜脈註射,模型組及正常組均給予等量L-DMEM液註射.移植後30 d,觀察4組大鼠一般情況、生存率、右心室收縮壓(RVSP)、右心室肥大指數(RVH)、血氣及肺小動脈的微觀結構變化.結果 移植後30 d,MMSC移植組及轉基因移植組大鼠的RVSP分彆為(30.2 ±2.1)和(29.2±1.1)mm Hg(1 mm Hg=0.133 kPa),RVH分彆為(37.9 ±3.2)%和(27.2±3.4)%,肺小動脈中膜厚度(MT)分彆為(21.3±3.4)和(14.3±2.8)μm,肺小動脈管腔橫截麵積與血管總橫截麵積的比值(V/T)分彆為(39.3 ±4.3)%和(43.0 ±1.5)%.上述指標與PH模型組相比差異均有統計學意義(P<0.01);其中MMSC移植組與轉基因移植組間RVH、MT及V/T的差異有統計學意義(P<0.05).另外,MMSC移植組與轉基因移植組動脈血氣各指標較PH模型組均明顯改善(P<0.05).同時觀察到肺內損傷血管脩複和血管新牛現象.結論 轉染有VEGF165基因的MMSC移植可有效減輕併逆轉野百閤堿誘導的PH進程,此作用較單純移植MMSC的治療效果更加明顯.其作用機製可能與血管脩複及血管新生有關.
목적 탐토혈관내피생장인자(VEGF)기인전염적골수간충질간세포(MMSC)이식치료폐동맥고압(PH)적료효화궤제.방법 체외분리배양SD대서MMSC,장함유증강형록색형광단백적VEGF질립전진MMSC.장SD대서수궤분위4조:정상대조조、PH모형조;MMSC이식조,전기인이식조.일차성항배부피하주사야백합감(50 mg/kg),복제PH모형,관찰21 d후,MMSC이식조급전기인이식조분별급여농도위5×106개세포/ml화5×105개세포/ml적MMSC현액1 ml정맥주사,모형조급정상조균급여등량L-DMEM액주사.이식후30 d,관찰4조대서일반정황、생존솔、우심실수축압(RVSP)、우심실비대지수(RVH)、혈기급폐소동맥적미관결구변화.결과 이식후30 d,MMSC이식조급전기인이식조대서적RVSP분별위(30.2 ±2.1)화(29.2±1.1)mm Hg(1 mm Hg=0.133 kPa),RVH분별위(37.9 ±3.2)%화(27.2±3.4)%,폐소동맥중막후도(MT)분별위(21.3±3.4)화(14.3±2.8)μm,폐소동맥관강횡절면적여혈관총횡절면적적비치(V/T)분별위(39.3 ±4.3)%화(43.0 ±1.5)%.상술지표여PH모형조상비차이균유통계학의의(P<0.01);기중MMSC이식조여전기인이식조간RVH、MT급V/T적차이유통계학의의(P<0.05).령외,MMSC이식조여전기인이식조동맥혈기각지표교PH모형조균명현개선(P<0.05).동시관찰도폐내손상혈관수복화혈관신우현상.결론 전염유VEGF165기인적MMSC이식가유효감경병역전야백합감유도적PH진정,차작용교단순이식MMSC적치료효과경가명현.기작용궤제가능여혈관수복급혈관신생유관.
Objective To explore the therapeutic effect and the mechanism of marrow mesenchymal stem cells (MMSCs) transfected with vascular endothelial growth factor ( VEGF) gene in the treatment of pulmonary hypertension in rats. Methods MMSCs from the bone marrow of Sprague-Dawley rats were isolated, cultured and propagated in vitro. pIRES2-EGFP-VEGF165 was transfected into MMSC. The healthy male SD rats were divided randomly into 4 groups: normal control group, pulmonary hypertension model group, MMSCs transplantation group and transfer gene transplantation group. A single subcutaneous monocrotaline (50 mg/kg) was injected to induce the model of pulmonary hypertension. The normal control group received a single subcutaneous dose of L-DMEM (low glucose Dulbecco's modified Eagle's medium).All four groups of rats were fed similarly. At Day 21 post-modeling, 5 × 106 MMSCs in 1 ml L-DMEM were injected into the MMSC group. 5 × 105 MMSC transfected by pIRES2-EGFP-VEGF165 were injected into the gene transplantation group. A same volume L-DMEM solution was also injected into the pulmonary hypertension model group and normal control group. The parameters of right ventricular systolic pressure (RVSP) , right ventricular hypertrophy index, blood gas analysis and microstructure as well as pulmonary microvascular changes were observed after 30 days. Results At Day 30 post-transplantation of MMSCs, the outcomes were as follows; RVSP was (30. 2 ±2. 1) and (29. 2 ± 1. 1) mm Hg (1 mm Hg =0. 133 kPa) in the MMSCs transplantation and gene transplantation groups respectively. The right ventricular hypertrophy indices were (37. 9 ±3. 2) % and (27. 2 ±3.4)% respectively. The media thickness of pulmonary artery ( MT) was (21.3 ± 3. 4 ) and ( 14. 3 ± 2. 8 ) μm respectively. The ratios of vascular area to total arterial area (V/T) were (39.3 ±4.3) % and (43.0±1.5) % respectively. As compare with the pulmonary hypertension model group, the above parameters were of statistical significances (P <0. 01). A comparison of right ventricle hypertrophy index, MT and V/T was of statistical significance between MMSC and gene transplantation groups ( P < 0. 05 ) . The blood gas analysis of the MMSCs transplantation and gene transplantation groups were better than the pulmonary hypertension mode group. Ultramicrostructure showed that neovascularization and small pulmonary arterial repair appeared in two transplantation groups. Conclusion MMSCs transfected by pIRES2-EGFP-VEGF165 transplantation may improve and reverse the MCT-induced progress of pulmonary hypertension in rats. And it is better than the MMSC transplantation. The potential mechanism is through arterial repair and neovascularization.
