CAJ | 학술논문

目的对脂多糖(LPS)诱导的脓毒症小鼠心功能进行观察,探讨LPS诱导脓毒症小鼠心功能障碍的病理生理机制.方法选取8周龄的雄性C57BL/6小鼠共60只随机(随机数字法)分为4组,对照组(n=15)和实验组共3组(n=15).对照组给予生理盐水腹腔注射(10 mg/kg);实验组行LPS腹腔注射(10 mg/kg),分别于6、12、24 h后通过超声观察各组小鼠心功能(n=12).取小鼠心、肾、肺组织包埋后HE染色(n=6),鉴定模型,通过免疫组化法(n=3)检测各组心脏组织PECAM-1、α-SMA的表达.运用RT-PCR技术检测小鼠心肌血管内皮生长因子(VEGF)和低氧诱导因子(HIF-1 α)表达水平,采用Western blot方法检测小鼠心脏p53与HIF-1α表达量,用ELISA法检测肿瘤坏死因子(TNF-α)和白介素(IL-6)的表达水平.所得数据采用独立样本t检验和单因素方差分析.结果实验组小鼠心脏左室收缩期前壁厚度、左室舒张期前壁厚度、左心室舒张期内径增加,每搏输出量降低,与对照组比较差异具有统计学意义(P＜0.05).通过免疫组化发现,实验组小鼠心脏中新生血管数量较对照组增加(P＜0.05);RT-PCR示实验组小鼠心脏中VEGF、HIF-1α表达量增高(P＜0.05);Western blot示实验组HIF-1α、p53的表达水平较对照组明显增高(P＜0.05);ELISA检测结果显示实验组VEGF、TNF-α等细胞因子表达水平较对照组增高(P＜0.05).结论脂多糖可导致小鼠心功能障碍,在此过程中心肌血管再生及细胞凋亡并存,VEGF和HIF-1α等参与了血管再生,而BAX、p53等参与了细胞凋亡.
목적 대지다당(LPS)유도적농독증소서심공능진행관찰,탐토LPS유도농독증소서심공능장애적병리생리궤제.방법 선취8주령적웅성C57BL/6소서공60지수궤(수궤수자법)분위4조,대조조(n=15)화실험조공3조(n=15).대조조급여생리염수복강주사(10 mg/kg);실험조행LPS복강주사(10 mg/kg),분별우6、12、24 h후통과초성관찰각조소서심공능(n=12).취소서심、신、폐조직포매후HE염색(n=6),감정모형,통과면역조화법(n=3)검측각조심장조직PECAM-1、α-SMA적표체.운용RT-PCR기술검측소서심기혈관내피생장인자(VEGF)화저양유도인자(HIF-1 α)표체수평,채용Western blot방법검측소서심장p53여HIF-1α표체량,용ELISA법검측종류배사인자(TNF-α)화백개소(IL-6)적표체수평.소득수거채용독립양본t검험화단인소방차분석.결과 실험조소서심장좌실수축기전벽후도、좌실서장기전벽후도、좌심실서장기내경증가,매박수출량강저,여대조조비교차이구유통계학의의(P＜0.05).통과면역조화발현,실험조소서심장중신생혈관수량교대조조증가(P＜0.05);RT-PCR시실험조소서심장중VEGF、HIF-1α표체량증고(P＜0.05);Western blot시실험조HIF-1α、p53적표체수평교대조조명현증고(P＜0.05);ELISA검측결과현시실험조VEGF、TNF-α등세포인자표체수평교대조조증고(P＜0.05).결론 지다당가도치소서심공능장애,재차과정중심기혈관재생급세포조망병존,VEGF화HIF-1α등삼여료혈관재생,이BAX、p53등삼여료세포조망.
Objective To study about the cardiac function of the mice suffering from sepsis induced by lipopolysaccharide (LPS) so as to probe the physiopathologic mechanism of the cardiac dysfunction of the mice.Method Sixty male C57BL/6 mice of eight weeks old were randomly (random number) divided into four groups:one control group (n =15) and three experimental groups (n =15 in each group).The mice of control group received intra-peritoneal injection of normal saline (10 mg/kg) while the mice of experimental groups got intra-peritoneal injection of LPS (10 mg/kg).The cardiac function of mice (n =12) was determined by echocardiography 6 h,12 h and 24 h later,respectively.The heart,kidney and lung tissues of mice (n =6) were stained with Haematoxylin-Eosin (HE) staining after embedding with paraffin for observing the histopathological changes under optic microscopy.The expressions of PECAM-1 and α-SMA of the heart tissue of mice (n =3) in three groups determined by immunohistochemical method.The RT-PCR method was used to test the expressions of VEGF (vascular endothelial growth factor) and HIF-α (hypoxia-inducible factor) of the myocardium of mice.In addition,the Western blot method was employed to test the levels of p53 and HIF-1α proteins in myocardium of mice,while ELISA was utilized to detect the level of tumor necrosis factor-α (TNF-α) and the interleukin-6 (IL-6).The data were analyzed by independent samples of t-test and one-way ANOVA respectively.Results The experiment result proved that the thickness of anterior wall of left ventricle of mice during systolic and diastolic periods increased and the inner diameter of the left ventricle also increased during the diastolic period in mice of the experimental group,while the stroke volume decreased compared with the control group (P ＜ 0.05).The immunohistochemical method showed that the new vessels of the mice' s heart in experimental groups increased compared with the control group (P ＜ 0.05).RT-PCR showed the expressions of VEGF and HIF-1α of the mice heart of experimental group increased (P ＜ 0.05) and Western blot showed the levels of HIF-1 α and p53 proteins in experimental groups increased significantly compared with the control group.The experimental group had higher levds of VEGF,HIF-1α,and IL-6 were evidenced by using ELISA than those of the control group (P ＜ 0.05).Conclusions The lipopolysaccharide can lead to cardiac dysfunction.In this process,myocardium angiogenesis and apoptosis phenomenon coexists,as VEGF and HIF-1α participating in angiogenesis,whereas BAX and p53 playing a role in the process of apoptosis.