抑制MLK3对铁过载肾衰竭小鼠心肌纤维化的作用观察  

Effect of inhibition of MLK3 on myocardial fibrosis in iron overload mice with renal failure

作　　者：盛丹丹 王琳 SHENG Dandan;WANG Lin(School of Medicine,Jiangsu University,Zhenjiang 212013,China)

机构地区：[1]江苏大学医学院,江苏镇江212013 [2]江苏大学附属医院心内科

出　　处：《山东医药》2023年第14期12-15,共4页Shandong Medical Journal

基　　金：国家自然科学基金项目(81801664)。

摘　　要：目的探讨抑制混合谱系激酶3(MLK3)对铁过载肾衰竭小鼠心肌纤维化的作用。方法将75只小鼠随机分为肾衰竭模型组60只和假手术组15只,肾衰竭模型组采用5/6肾切除术构建肾衰竭模型,假手术组仅切开背部皮肤后缝合。将造模成功的肾衰竭模型组小鼠分为肾衰竭组、肾衰竭铁过载组、MLK3抑制组、MLK3恢复组,每组各15只。肾衰竭铁过载组、MLK3抑制组、MLK3恢复组均给予0.2 mg/g右旋糖酐铁腹腔注射构建铁过载模型。MLK3抑制组于末次注射右旋糖酐铁后,给予MLK3抑制剂腹腔注射;MLK3恢复组先给予MLK3抑制剂灌胃,2周后给予尾静脉注射MLK3腺相关病毒。小鼠处死,取心脏组织,采用普鲁士蓝染色法检查心脏组织铁沉积情况;采用HE染色法观察心脏组织结构改变;采用Masson染色法观察心脏组织纤维化情况;采用免疫荧光法检测心脏组织纤维化相关蛋白波形蛋白(Vimentin)表达。结果普鲁士蓝染色显示,假手术组和肾衰竭组心脏组织无明显铁沉积,铁过载组心脏组织铁沉积明显增多。HE染色显示,假手术组和肾衰竭组心肌纤维排列整齐,细胞结构完整;铁过载组心肌纤维排列紊乱,结构破坏,细胞着色不均匀;MLK3抑制组心肌组织纤维排列较铁过载组整齐,但偶有少许纤维破裂;与MLK3抑制组比较,MLK3恢复组心肌纤维排列杂乱无章,断裂痕迹明显。Masson染色显示,铁过载组有明显蓝色胶原纤维;与铁过载组比较,MLK3抑制组蓝色胶原纤维沉积减少;与MLK3抑制组比较,MLK3恢复组蓝色胶原纤维明显增多。肾衰竭组心脏组织Vimentin表达较假手术组升高;与肾衰竭组比较,铁过载组Vimentin表达升高;与铁过载组比较,MLK3抑制组Vimentin表达降低;与MLK3抑制组比较,MLK3恢复组Vimentin表达升高(P<0.05或<0.01)。结论铁过载可能通过影响MLK3促进心肌纤维化的发生;抑制MLK3能够改善铁过载肾衰竭小鼠心肌纤维化程度。Objective To investigate the effect of inhibition of mixed lineage kinase 3(MLK3)on myocardial fibro⁃sis in renal failure mice induced by iron overload.Methods Seventy-five mice were randomly divided into the kidney failure model group of 60 and sham operation group of 15.In the kidney failure model group,we constructed the kidney failure models by 5/6 nephrectomy,while mice in the sham operation group only underwent incision of the back skin and suturing.The successfully modeled renal failure model mice were divided into renal failure group,renal failure iron over⁃load group,MLK3 inhibition group,and MLK3 activation group,with 15 mice in each group.Mice in the renal failure iron overload group,MLK3 inhibition group,and MLK3 activation group were all given 0.2 mg/g dextran iron through in⁃traperitoneal injection to build iron overload models.Mice in the MLK3 inhibition group received intraperitoneal injection of MLK3 inhibitor after the last injection of dextran iron.Mice in the MLK3 agonist group were first given an MLK3 inhibi⁃tor by gavage,and after 2 weeks,the MLK3 adeno-associated virus was injected into the tail vein.Mouse was euthanized and heart tissue was taken for examination of iron deposition in the heart tissue using Prussian blue staining method.The changes in cardiac tissue structure were observed by HE staining and the cardiac tissue fibrosis by Masson staining.The expression of fibrosis-related protein Vimentin in the heart tissues was detected by immunofluorescence assay.Results Prussian blue staining showed that there was no obvious iron deposition in the heart tissues of the sham operation group and the renal failure group,but the iron deposition in the heart tissues significantly increased in the iron overload group.HE staining showed that the myocardial fibers in the sham operation group and renal failure group were arranged neatly and the cell structure was intact.In the iron overload group,the arrangement of myocardial fibers was disordered,the structure was destroyed,and the cells

关 键 词：混合谱系激酶3 铁过载 肾衰竭 心肌纤维化 小鼠 

分 类 号：R332[医药卫生—人体生理学]