CAJ | 학술논문

目的研究急性心肌梗死时超声介导微泡造影剂携带肝细胞生长因子(HGF)基因靶向治疗的可行性.方法将54只雄性SD大鼠建立心肌梗死模型并随机分成3组(每组18只):US/MB-HGF组,予超声照射及尾静脉注射携带HGF质粒基因(pcDNA3.1-HGF)的微泡造影剂;US-HGF组,予超声照射及尾静脉注射PCDNA3.1-HGF,无微泡造影剂;US/MB-P组:即对照组,予超声照射及尾静脉注射携带空质粒的微泡造影剂.并按大鼠被处死的时间顺序,将每组分为3个亚组(每组6只):24 h组、7 d组和14 d组,分别取心肌组织做石蜡切片并做免疫组织化学染色,包括抗凋亡蛋白(bcl-2)的表达、HGF蛋白表达和CD34标记的毛细血管计数情况.结果 (1)心肌组织HGF蛋白表达:US/MB.HGF7 d亚组有明显的HGF蛋白表达,24 h组未见有明显的HGF的表达颗粒.US-HGF7d和14 d亚组可见少量HGF表达.US/MB-P组各亚组均未见HGF表达.(2)CDM标记的毛细血管计数:US/MB-HGF 14 d亚组为(367.6±17.6)个/高倍视野,明显高于US-HGF组[(268.9±0.8)个/高倍视野]和US/MB-P组[(186.8±11.8)个/高倍视野,均P＜0.05],各组24 h和7 d亚组的毛细血管计数均低于其14 d亚组.(3)梗死区bcl-2的表达:US/MB-HGF 7 d亚组为9.9%±0.5%,明显高于US-HGF组(6.3%±1.0%)和uS/MB-P组(3.5%±0.8%,均P＜0.05);US/MB-HGF 14 d亚组为6.7%±0.9%,明显高于US-HGF组(4.5%±0.8%)和US/MB-P组(2.1%±0.9%,均P＜0.05);3组24 h亚组均未见明显表达.结论超声介导造影剂微泡破裂可以使静脉注射的HGF基因在心肌局部浓度增高,增强其转染和表达,且可明显促进心肌新生血管的生长,减少急性梗死区心肌细胞的凋亡.
목적 연구급성심기경사시초성개도미포조영제휴대간세포생장인자(HGF)기인파향치료적가행성.방법 장54지웅성SD대서건립심기경사모형병수궤분성3조(매조18지):US/MB-HGF조,여초성조사급미정맥주사휴대HGF질립기인(pcDNA3.1-HGF)적미포조영제;US-HGF조,여초성조사급미정맥주사PCDNA3.1-HGF,무미포조영제;US/MB-P조:즉대조조,여초성조사급미정맥주사휴대공질립적미포조영제.병안대서피처사적시간순서,장매조분위3개아조(매조6지):24 h조、7 d조화14 d조,분별취심기조직주석사절편병주면역조직화학염색,포괄항조망단백(bcl-2)적표체、HGF단백표체화CD34표기적모세혈관계수정황.결과 (1)심기조직HGF단백표체:US/MB.HGF7 d아조유명현적HGF단백표체,24 h조미견유명현적HGF적표체과립.US-HGF7d화14 d아조가견소량HGF표체.US/MB-P조각아조균미견HGF표체.(2)CDM표기적모세혈관계수:US/MB-HGF 14 d아조위(367.6±17.6)개/고배시야,명현고우US-HGF조[(268.9±0.8)개/고배시야]화US/MB-P조[(186.8±11.8)개/고배시야,균P＜0.05],각조24 h화7 d아조적모세혈관계수균저우기14 d아조.(3)경사구bcl-2적표체:US/MB-HGF 7 d아조위9.9%±0.5%,명현고우US-HGF조(6.3%±1.0%)화uS/MB-P조(3.5%±0.8%,균P＜0.05);US/MB-HGF 14 d아조위6.7%±0.9%,명현고우US-HGF조(4.5%±0.8%)화US/MB-P조(2.1%±0.9%,균P＜0.05);3조24 h아조균미견명현표체.결론 초성개도조영제미포파렬가이사정맥주사적HGF기인재심기국부농도증고,증강기전염화표체,차가명현촉진심기신생혈관적생장,감소급성경사구심기세포적조망.
Objective To evaluate whether ultrasonic microbubble destruction (US/MB) could enhance the therapeutic effects of hepatocyte growth factor (HGF) gene transfer for acute myocardial infarction (MI).Methods MI was induced by left anterior descending artery ligation in male SD rats.Two to 4 hours thereafter,MI rats were randomly treated with tail vein infusing pc-DNA3.1-HGF plasmid mixed with microbubbles (US/MB-HGF group,n=18);tail vein infusing pc-DNA3.1-HGF plasmid mixed with saline (US-HGF group,n=18);tail vein infusing empty plasmid mixed with micrnhubbles (US/MB-Pgroup,n=18).All rats were exposed to ultrasound treatment thereafter till contrast imaging disappeared in cardiac region.Rats were sacrificed at 24 hours,7 days or 14 days,respectively (n=6 each) and myocardial protein expression of bcl-2 and HGF as well as microvascular density (MVD) were determined.Results The myocardial protein expressions of bcl-2 and HGF in US/MB-HGF group were significantly higher than those in US-HGF and US/MB-P groups at 7 days post MI (all P＜0.01) and MVD was significantly higher in US/MB-HGF group (367.6 ±17.6) than that in US-HGF (268.9±0.8) and US/MB-P (186.8±11.8) groups (all P＜0.05) at 14 days post MI.Conclusions Ultrasound-mediated microbubble destruction could enhance systemic HGF administration induced myocardial angiogenesis and reduce systemic HGF administration induced myocardial apoptosis in rats with acute MI.