脂联素通过下调Nox2表达减轻急性酒精性心肌损伤

发布时间：2018-05-27 01:24

本文选题：酒精 + 心肌损伤　；参考：《山西医科大学》2017年硕士论文

【摘要】：目的探讨脂联素(APN)对小鼠急性酒精性心肌损伤的影响并研究相关机制,为急性酒精性心肌损伤及酒精性心肌病的预防和诊治提供实验依据及新思路。方法取20只健康8周龄SPF级C57BL/6J雄性小鼠,随机分为正常实验组(n=10只)与正常模型组(n=10只)。将8周龄纯合子脂联素基因敲除(APN-/-)雄性鼠(SPF级、近交系C57BL/6J,20只)随机分为APN-/-对照组(n=10只)与APN-/-模型组(n=10只);各模型组均给予腹腔注射乙醇3g/Kg(体重)·d,各对照组均给予腹腔注射相等量的生理盐水。4组小鼠均正常摄食、饮水,连续3天后进行各项指标检测:观察小鼠一般情况;用动物超声机及小动物专用探头测定心脏结构参数及相关功能指标;检测血清乳酸脱氢酶(LDH)、血浆N尾端B型脑钠肽原(NT-proBNP);检测心肌组织Casepase-3活力及TUNEL检测心肌组织细胞凋亡;制作心肌组织HE染色及Masson染色切片;制备心肌匀浆,测定MDA、ROS含量及SOD活性;Western blot法测定心肌组织Nox2蛋白表达情况。结果与正常对照组相比,正常模型组射血分数显著下降(P0.01);血清LDH、血浆NT-proBNP浓度分别升高了1.98倍、5.13倍(均P0.01);HE染色及Masson染色结果示心肌结构紊乱、心肌纤维走向迂曲、断裂,胶原纤维增多,CVF值升高2.63倍(P0.01);心肌组织内Casepase-3活性升高了2.58倍(P0.01),TUNEL法示阳性凋亡心肌细胞增多了12.67倍(P0.01);心肌组织内ROS含量、MDA含量分别升高了1.68倍、2.87倍(均P0.01),SOD活性升高2.92倍(P0.01);Nox2蛋白表达量升高1.87倍(P0.01)。与APN-/-对照组比较,APN-/-模型组射血分数显著下降(P0.01),其余指标均显著升高(均P0.01)。与正常模型组比较,APN-/-模型组小鼠射血分数降低更为显著(P0.01),血清LDH、NT-proBNP浓度分别升高了1.30倍、1.25倍(均P0.01);HE染色及Masson染色示心肌结构紊乱无序、心肌纤维走向迂曲、断裂更加明显、胶原纤维增多明显,CVF值升高了1.55倍(P0.01);心肌组织内Casepase-3活性升高1.66倍(P0.01),TUNEL法示阳性凋亡心肌细胞比例升高1.64倍(P0.01);心肌组织内ROS含量、MDA含量分别升高1.42倍、1.39倍(均P0.01),SOD活性降低28%(P0.01),Nox2蛋白表达量升高1.44倍(P0.01)。结论脂联素(APN)可以减轻小鼠急性酒精性心肌损伤,其机制与脂联素抑制心肌组织Nox2表达发挥抗氧化应激作用有关。
[Abstract]:Objective to investigate the effect of APN on acute alcoholic myocardial injury in mice and its related mechanism, and to provide experimental basis and new ideas for the prevention and treatment of acute alcoholic myocardial injury and alcoholic cardiomyopathy. Methods Twenty healthy 8-week-old SPF grade C57BL/6J male mice were randomly divided into normal experimental group (n = 10) and normal model group (n = 10). The 8 week old homozygous adiponectin gene knockout APN / -) male mice with SPF grade, Inbred line C57BL / 6 JJ (20) were randomly divided into APN-r-control group (n = 10) and APN-r-model group (n = 10). Each model group was given ethanol 3 g / Kg (body weight) d, and each control group was given a normal diet and drinking water of the same amount of normal saline. After 3 days of continuous testing, the general situation of mice was observed, the parameters of heart structure and related functional indexes were measured by animal ultrasound machine and small animal special probe. Serum lactate dehydrogenase (LDH), plasma N-tail B-type brain natriuretic peptide (NT-proBNPN), myocardial tissue Casepase-3 activity and TUNEL detection of myocardial apoptosis, myocardial HE staining and Masson staining sections were made, and myocardial homogenate was prepared. The content of MDAA Ros and the expression of Nox2 protein in myocardium were determined by Western blot. Results compared with the normal control group, the ejection fraction of the normal model group decreased significantly (P 0.01), the serum LDH and plasma NT-proBNP concentration increased by 1.98 times and 5.13 times respectively (P 0.01 he staining and Masson staining showed that the myocardial structure was disordered, the myocardial fibers were tortuous and broken. The Casepase-3 activity in myocardial tissue increased by 2.58 times and the number of positive apoptotic cardiomyocytes increased by 12.67 times by Tunel method. The content of ROS in myocardial tissue was increased by 1.68 times and 2.87 times respectively (all P0.01SOD activity increased). The protein expression of Nox2 was 1.87 times higher than that of P0.01. Compared with the control group, the ejection fraction of the APN-p-model group decreased significantly (P 0.01), and the other indexes increased significantly (P 0.01). Compared with the normal model group, the ejection fraction decreased significantly in the APN-r-model group, and the serum LDHNT-proBNP concentration increased by 1.30 times and 1.25 times respectively (P 0.01 he staining and Masson staining showed that the myocardial structure was disordered, the myocardial fiber direction was tortuous, and the rupture was more obvious. Casepase-3 activity in myocardial tissue increased by 1.66 times and the percentage of positive apoptotic cardiomyocytes was increased by 1.64 times by Tunel method. The content of ROS in myocardial tissue was increased by 1.42 times and 1.39 times respectively (all P0.01SOD activity). The expression of P0.01Nox2 protein increased by 1.44 times. Conclusion adiponectin (APN) can attenuate acute alcoholic myocardial injury in mice, and its mechanism is related to the antioxidation stress effect of adiponectin on the expression of Nox2 in myocardium.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R542.2

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