中华麻醉学杂志
中華痳醉學雜誌
중화마취학잡지
CHINESE JOURNAL OF ANESTHESIOLOGY
2012年
10期
1252-1256
,共5页
林群%雷立华%林财珠%曾邦雄%梁富球%林献忠%郑辉哲%蔡宏达%高友光%杨庆
林群%雷立華%林財珠%曾邦雄%樑富毬%林獻忠%鄭輝哲%蔡宏達%高友光%楊慶
림군%뢰립화%림재주%증방웅%량부구%림헌충%정휘철%채굉체%고우광%양경
肝细胞生长因子%间质干细胞移植%生物,基因修饰%高血压,肺性
肝細胞生長因子%間質榦細胞移植%生物,基因脩飾%高血壓,肺性
간세포생장인자%간질간세포이식%생물,기인수식%고혈압,폐성
Hepatocyte growth factor%Mesenchymal stem cell transplantation%Organisms,genetically modified%Hypertension,pulmonary
目的 评价人肝细胞生长因子(hHGF)基因修饰对骨髓间质于细胞(MSCs)移植改善重度肺动脉高压大鼠肺微血管稀薄作用的影响.方法 将原代培养的F344大鼠MSCs转导携带HGF基因的慢病毒载体和空载体,以获得HGF-MSCs和EGFP-MSCs.7周龄近交系雄性F344大鼠,体重180 ～ 250 g,采用野百合碱复合单肺切除法建立重度肺动脉高压大鼠模型.取重度肺动脉高压大鼠66只,采用随机数字表法,将其随机分为3组(n=22):对照组(C组)、空载体慢病毒转导的MSCs(EGFP-MSCs)组(E组)和hHGF转导的MSCs (hHGF-MSCs)组(H组).C组经颈外静脉注射DMEM1ml,E组注射含5× 105个EGFP-MSCs的DMEM 1 ml和H组注射含5×105个hHGF-MSCs的DMEM 1ml.注射野百合碱后45 d内进行生存分析；颈外静脉注射后2周,记录平均肺动脉压(MPAP),计算有心室肥厚指数,观察肺组织MSCs分布情况,测定肺组织鼠肝细胞生长因子(rHGF)和hHGF蛋白的含量,测定Ⅷ因子的表达情况以计算肺微血管密度,并行肺动脉钡剂灌注造影.结果 C组和E组肺组织均未检测出hHGF.与C组比较,E组和H组MPAP和右心室肥厚指数降低,肺组织rHGF蛋白含量、肺微血管密度和生存率升高(P＜0.01).与E组比较,H组MPAP和右心室肥厚指数降低,肺组织rHGF蛋白含量、肺微血管密度和生存率升高(P＜0.01).E组和H组肺内均可见大量绿色荧光的MSCs分布.肺动脉钡剂灌注造影显示,H组末梢血管灌注背景明显多于E组和C组.结论 hHGF基因修饰可促进MSCs移植改善重度肺动脉高压大鼠肺微血管稀薄的作用.
目的 評價人肝細胞生長因子(hHGF)基因脩飾對骨髓間質于細胞(MSCs)移植改善重度肺動脈高壓大鼠肺微血管稀薄作用的影響.方法 將原代培養的F344大鼠MSCs轉導攜帶HGF基因的慢病毒載體和空載體,以穫得HGF-MSCs和EGFP-MSCs.7週齡近交繫雄性F344大鼠,體重180 ～ 250 g,採用野百閤堿複閤單肺切除法建立重度肺動脈高壓大鼠模型.取重度肺動脈高壓大鼠66隻,採用隨機數字錶法,將其隨機分為3組(n=22):對照組(C組)、空載體慢病毒轉導的MSCs(EGFP-MSCs)組(E組)和hHGF轉導的MSCs (hHGF-MSCs)組(H組).C組經頸外靜脈註射DMEM1ml,E組註射含5× 105箇EGFP-MSCs的DMEM 1 ml和H組註射含5×105箇hHGF-MSCs的DMEM 1ml.註射野百閤堿後45 d內進行生存分析；頸外靜脈註射後2週,記錄平均肺動脈壓(MPAP),計算有心室肥厚指數,觀察肺組織MSCs分佈情況,測定肺組織鼠肝細胞生長因子(rHGF)和hHGF蛋白的含量,測定Ⅷ因子的錶達情況以計算肺微血管密度,併行肺動脈鋇劑灌註造影.結果 C組和E組肺組織均未檢測齣hHGF.與C組比較,E組和H組MPAP和右心室肥厚指數降低,肺組織rHGF蛋白含量、肺微血管密度和生存率升高(P＜0.01).與E組比較,H組MPAP和右心室肥厚指數降低,肺組織rHGF蛋白含量、肺微血管密度和生存率升高(P＜0.01).E組和H組肺內均可見大量綠色熒光的MSCs分佈.肺動脈鋇劑灌註造影顯示,H組末梢血管灌註揹景明顯多于E組和C組.結論 hHGF基因脩飾可促進MSCs移植改善重度肺動脈高壓大鼠肺微血管稀薄的作用.
목적 평개인간세포생장인자(hHGF)기인수식대골수간질우세포(MSCs)이식개선중도폐동맥고압대서폐미혈관희박작용적영향.방법 장원대배양적F344대서MSCs전도휴대HGF기인적만병독재체화공재체,이획득HGF-MSCs화EGFP-MSCs.7주령근교계웅성F344대서,체중180 ～ 250 g,채용야백합감복합단폐절제법건립중도폐동맥고압대서모형.취중도폐동맥고압대서66지,채용수궤수자표법,장기수궤분위3조(n=22):대조조(C조)、공재체만병독전도적MSCs(EGFP-MSCs)조(E조)화hHGF전도적MSCs (hHGF-MSCs)조(H조).C조경경외정맥주사DMEM1ml,E조주사함5× 105개EGFP-MSCs적DMEM 1 ml화H조주사함5×105개hHGF-MSCs적DMEM 1ml.주사야백합감후45 d내진행생존분석；경외정맥주사후2주,기록평균폐동맥압(MPAP),계산유심실비후지수,관찰폐조직MSCs분포정황,측정폐조직서간세포생장인자(rHGF)화hHGF단백적함량,측정Ⅷ인자적표체정황이계산폐미혈관밀도,병행폐동맥패제관주조영.결과 C조화E조폐조직균미검측출hHGF.여C조비교,E조화H조MPAP화우심실비후지수강저,폐조직rHGF단백함량、폐미혈관밀도화생존솔승고(P＜0.01).여E조비교,H조MPAP화우심실비후지수강저,폐조직rHGF단백함량、폐미혈관밀도화생존솔승고(P＜0.01).E조화H조폐내균가견대량록색형광적MSCs분포.폐동맥패제관주조영현시,H조말소혈관관주배경명현다우E조화C조.결론 hHGF기인수식가촉진MSCs이식개선중도폐동맥고압대서폐미혈관희박적작용.
Objective To investigate the effect of human hepatocyte growth factor (hHGF) genetic modification on the ameliorating effects of mesenchymal stem cells (MSCs) implantation on pulmonary microvascular rarefaction in a rat model of pulmonary hypertension (PH).Methods MSCs were obtained from F344 rats and transduced with lentiviral vector modified with human HGF (hHGF-MSCs) or empty vector (EGFP-MSCs).Sixty-six 7 week old male F344 rats weighing 180-250 g were used in this study.PH was induced by left pneumonectomy and subcutaneous monocrotaline (MCT) 60 mg/kg injected at 2 weeks after operation.The animals with PH were randomly divided into 3 groups:control group (group C),EGFP-MSCs group (group E) and HGF-MSCs group (group H).Groups H and E received hHGF-MSCs or EGFP-MSCs 5 × 105 in DMEM 1 ml iv at 3 weeks after subcutaneous MCT injection,while group C received plain DMEM 1 ml.Mean pulmonary arterial pressure (mPAP) was measured and right ventricular hypertrophy and angiogenesis in the lung were assessed and the content of rat HGF (rHGF) and hHGF protein in lung tissue and pulmonary capillary density (by immuno-histochemistry) was measured at 2 weeks after MSCs implantation.The survival rates within 45 days after MCT administration were compared among the 3 groups.Results No hHGF was detected in groups C and E.Both hHGF-MSCs and EGFP-MSCs significantly reduced MPAP and right ventricular hypertrophy and increased pulmonary capillary density and survival rates in groups H and E as compared with group C and the efficacy of hHGF-MSCs was significantly greater than that of EGFP-MSCs.Barium angiography revealed that distal pulmonary vasculature was significantly increased in group H as compared with groups E and C.The survival of the rats receiving hHGF-MSCs was significantly longer in group H than that in groups E and C.Conclusion hHGF genetic modification can improve the ameliorating effects of MSCs implantation on PH-related microvascular rarefaction.
