压力超负荷诱导的心肌肥厚/心力衰竭模型鼠肝肾损伤及巨噬细胞活化水平的变化

权月; 田格尔; 周骏腾; 吴文超; 刘小菁

doi:10.12182/20200560606

压力超负荷诱导的心肌肥厚/心力衰竭模型鼠肝肾损伤及巨噬细胞活化水平的变化

The Observation of Liver and Kidney Injury and the Activation of Macrophages in the Overload Pressure Induced Cardiac Hypertrophy/Heart Failure Mouse Model

摘要

摘要:
目的观察压力超负荷诱导的心肌肥厚/心力衰竭小鼠模型中肝肾组织功能有无损伤，以及在此过程中巨噬细胞活化水平的变化。
方法 C57BL/6小鼠通过胸主动脉缩窄术(transverse aortic constriction，TAC)构建压力超负荷诱导的心肌肥厚/心力衰竭小鼠模型，假手术组小鼠主动脉弓不进行结扎。在TAC术后4周（4周TAC组）及8周（8周TAC组），各组小鼠分别进行超声心动图检查、采血及处死小鼠后取小鼠心、肝脏、肾脏组织标本，检测小鼠肝功能〔包括丙氨酸氨基转移酶（alanine aminotransferase, ALT）、门冬氨酸氨基转移酶（aspartate aminotransferase, AST）、总胆红素（total bilirubin, TBil）〕，以及肾功能〔血肌酐（serum creatinine, Scr）〕，小鼠心、肝、肾组织苏木精-伊红染色(hematoxylin-eosin staining，HE)观察病理形态学的改变，免疫组织化学染色法检测心脏、肝、肾组织中巨噬细胞活化的标志物F4/80蛋白的表达变化。
结果心脏超声检测结果显示，与假手术组相比，4周、8周TAC组小鼠心功能指标中左室舒张末期后壁厚度(left ventricular end-diastolic posterior wall thickness, LVPWd)、左室内径(left ventricular internal diameter in diastole, LVIDd)升高，左室射血分数（left ventricular ejection fraction, EF%）和左室短轴缩短率（left ventricular fractional shortening, FS%）均下降（P<0.05）。与假手术组比较，4周、8周TAC组小鼠血ALT、AST、TBil及Scr含量均升高（P <0.05）；HE染色显示4周TAC组小鼠肝组织即出现空泡化、肝窦轻度充血、炎性浸润等病理改变，8周TAC组小鼠肝损伤更加显著。TAC小鼠肾组织亦有轻微损伤表现，如肾小球轻度损伤，轻微出血。F4/80蛋白免疫组织化学染色结果显示，与Sham组比较，4周及8周TAC组小鼠心脏、肝脏的巨噬细胞活化增强（P <0.005），而肾脏中巨噬细胞活化水平无明显差异。
结论压力超负荷诱导的心肌肥厚/心力衰竭小鼠有肝肾损伤发生，此过程有巨噬细胞参与。

Abstract:
Objective The purpose of this study is to investigate the injury of liver and kidney tissues in overload pressure induced cardiac hypertrophy/heart failure mice model and the changes of macrophage activation level.
Methods 6-8 week-old C57BL/6 mice were subjected to transverse aortic constriction (TAC) surgery to establish the cardiac hypertrophy/heart failure mouse model induced by pressure overload, while the aortic was not ligated in the Sham group. At 4 weeks and 8 weeks after TAC, the mice of each group were subjected to echocardiography and blood collection. And mice were sacrificed to collect samples of the heart, liver, and kidney tissues. The contents of plasma alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBil) and serum creatinine (Scr) in Sham group and two operation groups were determined. The histological changes of liver, heart and kidney tissues were observed by HE staining, and the expression of the marker of macrophage activation, F4/80 protein, was detected in the heart, liver and kidney tissue by immunohistochemical staining.
Results Cardiac hypertrophy occurred at 4 weeks after TAC operation in C57BL/6 mice and developed into heart failure at 8 weeks after TAC. The echocardiography showed that, compared with the Sham group, the left ventricular end-diastolic posterior wall thickness (LVPWd) and the left ventricular internal diameter in diastole (LVIDd) were significantly increased, while the left ventricular ejection fraction (EF) and the left ventricular fractional shortening (FS) were significantly decreased (P<0.05) in the 4-week-TAC group and 8-week-TAC group. The plasma content of ALT, AST, TBil and Scr in the 4-week-TAC group and 8-week-TAC group were significantly higher than those in the Sham group (P<0.05). HE staining showed obvious liver pathological changes in TAC mice, such as vacuolation, mild hepatic sinusoid congestion and inflammatory infiltration in mice post 4 weeks after surgery, and such liver injury was worse in mice post 8 weeks after surgery. Besides, there was a slight damage in renal tissue shown by HE staining, such as slight glomerular injury and slight bleeding. F4/80 protein immunohistochemical staining results demonstrated that the activation of macrophages in the heart and liver in the 4-week-TAC group and 8-week-TAC group was significantly increased than that in the sham group (P<0.05), but there was no significant difference in kidney tissues in groups.
Conclusion Macrophages are involved in the process of liver and kidney injury in cardiac hypertrophy/heart failure.

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