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双侧颈上神经节切除对压力负荷型心力衰竭小鼠心室重构及心功能的影响

Bilateral superior cervical ganglionectomy attenuates cardiac remodeling and improves cardiac function in pressure-overloaded heart failure mice

摘要目的:探讨双侧颈上神经节切除对压力负荷型心力衰竭(心衰)小鼠心室重构及心功能的影响。方法:健康雄性6周龄C57BL/6小鼠共24只,体重20~25 g。采用随机数字法分为4组,即对照组( n=6,进行假手术)、去神经组( n=6,手术切除双侧颈上神经节)、心衰组[ n=6,采用重度胸主动脉缩窄术(sTAB)建立压力负荷型心衰模型]和心衰去神经组( n=6,于sTAB术后2周行双侧颈上神经节切除术)。分别于sTAB术前以及后第2、3、4周行超声心动图检测小鼠心功能。于sTAB术后第4周末处死小鼠,取心肌组织。心肌组织切片行苏木素-伊红(HE)、Masson和免疫组织化学染色进行心肌形态和组织学分析;提取心肌组织蛋白,采用Western blot法检测其酪氨酸羟化酶(TH)、B型利钠肽(BNP)及β1肾上腺素能受体(AR-β1)的蛋白表达水平。 结果:心衰组和心衰去神经组小鼠sTAB术后第2、3、4周左心室射血分数(LVEF)逐渐减低,术后第4周心衰去神经组小鼠LVEF高于心衰组( P<0.05),而去神经组小鼠各时间点LVEF与对照组比较差异均无统计学意义( P均>0.05)。sTAB术后第4周HE染色结果显示,与对照组比较,去神经组小鼠心肌细胞横截面积差异无统计学意义( P>0.05),心衰组小鼠则较大( P<0.05),而心衰去神经组小鼠较心衰组小( P<0.05)。Masson染色结果显示,对照组和去神经组小鼠心肌组织几乎无蓝染,而心衰组小鼠心肌组织出现大量蓝染,心衰去神经组小鼠蓝染组织少于心衰组。免疫组织化学染色结果显示,去神经组小鼠TH阳性神经纤维密度与对照组比较差异无统计学意义( P>0.05),而心衰组小鼠心肌组织中可见大量TH阳性的神经纤维,心衰去神经组小鼠心肌组织中TH阳性神经纤维的密度较心衰组低( P<0.05)。Western blot结果显示,心衰组小鼠心肌组织中TH蛋白表达水平高于对照组( P<0.05),而心衰去神经组则低于心衰组( P<0.05),且BNP蛋白在各组间表达的差异与TH蛋白相似。与对照组比较,去神经组小鼠AR-β1蛋白表达水平差异无统计学意义( P>0.05),心衰组则较低( P<0.05),而心衰去神经组与心衰组该指标差异无统计学意义( P>0.05)。免疫组织化学染色结果显示,与对照组比较,去神经组小鼠心脏CD68阳性细胞密度差异无统计学意义( P>0.05),心衰组则较高( P<0.05),而心衰去神经组较心衰组低( P<0.05)。 结论:切除双侧颈上神经节可减小心衰后小鼠心脏交感神经密度,减少巨噬细胞浸润,减轻心室重构,改善心功能。

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abstractsObjective:To investigate the effect of bilateral superior cervical ganglionectomy on cardiac remodeling and function in pressure-overloaded heart failure (HF) mice.Methods:Pressure-overloaded HF mouse model was produced by severe thoracic aorta banding (sTAB). Bilateral superior cervical ganglionectomy (SCGx) was performed 2 weeks after sTAB. Twenty four 6-week-old male C57BL/6 mice were randomized divided into 4 groups ( n=6 each): control group: sham sTAB+sham SCGx; denervated group: sham sTAB+SCGx; HF group: sTAB+sham SCGx; denervated HF group: sTAB+SCGx. Cardiac function was measured by echocardiography at week 0, 1, 2, and 4 after sTAB, respectively. All mice were sacrificed at the end of week 4 and heart tissues were harvested. HE and Masson staining were performed. Immunohistochemical staining (IHC) for tyrosine hydroxylase (TH), adrenergic receptor β1 (AR-β1) and CD68 was performed. Western blot was used to determine the protein expression level of TH, B type natriuretic peptide (BNP), and AR-β1. Results:Left ventricular ejection fraction (LVEF) declined continuously in HF group. LVEF was similar between denervated HF group and control group at various time points ( P>0.05). LVEF was significantly higher in denervated HF group than in HF group at the end of week 4 ( P<0.05). HE staining showed that cross sectional cardiomyocyte area was significantly larger in HF group than in control group and denervated HF group ( P<0.05), which was similar between denervated HF group and control group ( P>0.05). Masson staining showed that fibrosis level was significantly lower in denervated HF group than in HF group ( P<0.05). IHC showed that TH+nerves and CD68 + macrophages were significantly increased in HF mice as compared to control mice ( P<0.05), whereas this change was abolished in denervated HF group. AR-β1 was significantly down-regulated in HF group compared with control group ( P<0.05), which was not affected by denervation ( P>0.05). Western blot demonstrated that the expression level of TH and BNP was significantly higher in HF group compared with the control group ( P<0.05), whereas this difference was diminished in denervated HF group ( P>0.05). Conclusion:Bilateral superior cervical ganglionectomy can reduce sympathetic innervation and macrophage infiltration in pressure overloaded failure heart, thus attenuate cardiac remodeling and improve cardiac function.

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栏目名称 心力衰竭和心肌病
DOI 10.3760/cma.j.cn112148-20200603-00458
发布时间 2025-02-25
基金项目
国家自然科学基金 National Natural Science Foundation of China
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中华心血管病杂志

中华心血管病杂志

2021年49卷4期

345-352页

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