CAJ | 학술논문

目的对脓毒症小鼠发生心肌损伤时心肌血管再生和细胞凋亡情况及相关机制的研究.方法将40只8周左右的雄性C57BL/6小鼠随机(随机数字法)分为两组,实验组(n=20)和对照组(n=20),实验组用脂多糖(LPS)腹腔注射(10 mg/kg),对照组给以生理盐水(10mg/kg),6h后超声观察两组小鼠心功能(n=40),后取两组小鼠心脏、肺脏、肾脏组织石蜡包埋切片做HE染色(n=6)观察病理学变化鉴定模型,免疫组化(n=3)PECAM -1、α-SAM染色观察心肌血管再生,心肌细胞凋亡染色(TUNLE) (n=3)观察细胞凋亡情况,提取心脏组织RNA(n=6)通过RT-PCR技术对HIF-1α等血管再生因子进行检测,所得数据处理均采用独立样本t检验.结果实验组与对照组相比小鼠心脏左室舒张期前壁厚度(t=-4.60,P＜0.05)、左室收缩期前壁厚度增厚(t=-3.24,P＜0.05),左心室舒张期内径减小(t=3.57,P＜0.01),每搏输出量下降(t=5.51,P＜0.01),免疫组化染α-SAM抗体显示实验组心脏新生血管数增加(t=-11.00,P＜0.01),凋亡染色(TUNEL)存在心肌细胞凋亡[实验组比对照组:(191.31±5.41) vs.(52.24±4.32)],RT-PCR显示HIF-1α表达显著增高(t=-8.12,P＜0.05).结论脓毒症小鼠存在明显心肌血管再生、细胞凋亡及心功能障碍,导致血管再生的通路有低氧诱导因子( HIF -1α)的参与.
목적 대농독증소서발생심기손상시심기혈관재생화세포조망정황급상관궤제적연구.방법 장40지8주좌우적웅성C57BL/6소서수궤(수궤수자법)분위량조,실험조(n=20)화대조조(n=20),실험조용지다당(LPS)복강주사(10 mg/kg),대조조급이생리염수(10mg/kg),6h후초성관찰량조소서심공능(n=40),후취량조소서심장、폐장、신장조직석사포매절편주HE염색(n=6)관찰병이학변화감정모형,면역조화(n=3)PECAM -1、α-SAM염색관찰심기혈관재생,심기세포조망염색(TUNLE) (n=3)관찰세포조망정황,제취심장조직RNA(n=6)통과RT-PCR기술대HIF-1α등혈관재생인자진행검측,소득수거처리균채용독립양본t검험.결과 실험조여대조조상비소서심장좌실서장기전벽후도(t=-4.60,P＜0.05)、좌실수축기전벽후도증후(t=-3.24,P＜0.05),좌심실서장기내경감소(t=3.57,P＜0.01),매박수출량하강(t=5.51,P＜0.01),면역조화염α-SAM항체현시실험조심장신생혈관수증가(t=-11.00,P＜0.01),조망염색(TUNEL)존재심기세포조망[실험조비대조조:(191.31±5.41) vs.(52.24±4.32)],RT-PCR현시HIF-1α표체현저증고(t=-8.12,P＜0.05).결론 농독증소서존재명현심기혈관재생、세포조망급심공능장애,도치혈관재생적통로유저양유도인자( HIF -1α)적삼여.
Objective To investigate the angiogenesis,apoptosis and their mechanisms in septic mice with myocardial injury.Methods Forty male C57BL/6 mice aged 8 weeks were randomly ( random number) divided into two groups:the sepsis group and the control group.The mice of sepsis group were treated with lipopolysaccharide (LPS) ( 10 mg/kg Intraperitoneal injection) while the mice of control group were treated with saline solution instead (10 mg/kg Intraperitoneal injection).Cardiac function of mice (n =40) was evaluated with ultrasound 6 hours after LPS administration.Subsequently,the tissues of heart,lung and kidney of mice (n =6) were taken and treated with Haematoxylin -Eosin staining (H&E) in order to observe the pathological changes and verify the successfulness of modeling.Immunohistochemistry staining with PECAM - 1 and α - SMA was used to identify the angiogenesis in the heart ( n =3 ),while the TUNEL apoptosis assay was applied for detecting the myocardial cell apoptosis ( n =3 ).The mRNA was extracted from heart tissue (n =6) to observe the expression of HIF-1 ot which was proved to be an angiogenesis factor.All the results were analyzed by independent sample t - test.Results Compared to the control group,mice in the sepsis group showed increased in thickness of left ventricular diastolic anterior wall ( t =- 4.60,P ＜ 0.05 ) and thickness of left ventricular systolic anterior wall (t =-3.24,P ＜0.05 ) along with decrease in left ventricular end diastolic diameter ( t =3.57,P ＜ 0.01 ) and stroke volume ( t =5.51,P ＜ 0.01 ).Immunohistochemistry staining with alpha - SAM antibody revealed increase in cardiac angiogenesis in the sepsis group (t =- 11.00,P ＜ 0.01 ).TUNEL apoptosis assay demonstrated apoptosis of the cardiomyocytes [ sepsis group versus control group:( 191.31 ±5.41 ) vs ( 52.24 ±4.32) ] and RT - PCR showed an increase in the expression of HIF - 1 alpha in the mice of the sepsis group ( t =- 8.12,P ＜0.05) Conclusions There were apparent myocardial angiogenesis,apoptosis and cardiac dysfunction in septic animal models.HIF-1α might play a role in the angiogenesis pathway.