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2020, 10, v.43 1308-1311+1317
小鼠主动脉斑块大体油红O染色技术的改进
基金项目(Foundation): 新疆维吾尔自治区重点实验室开放课题项目(2017D04001)
邮箱(Email):
DOI:
投稿时间: 2020-01-10
投稿日期(年): 2020
修回时间: 2020-02-10
终审时间: 2020-10-15
终审日期(年): 2020
审稿周期(年): 1
发布时间: 2020-10-15
出版时间: 2020-10-15
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摘要:

目的对小鼠主动脉斑块大体油红O染色技术进行改进,以准确测定动脉粥样硬化小鼠主动脉壁斑块面积。方法选取普通饲料喂养10月龄雄性ApoE-/-小鼠行整根主动脉剥离,显微弹簧剪去除主动脉外膜脂肪并纵向剖开整根主动脉,不同温度梯度下(25℃、45℃、65℃)油红O染料孵育10 min,吸水纸上铺展主动脉,用图像分析软件测定主动脉各部位的斑块面积。结果不同温度油红O染色发现,45℃较25℃孵育动脉斑块着色明显,呈血红色,易于测量,而65℃孵育条件下,斑块出现脱落现象,故45℃为最佳温度。油红O染色显示ApoE-/-小鼠的主动脉弓、胸主动脉、腹主动脉以及髂总动脉均可见明显的粥样硬化斑块,大体主动脉斑块面积为(29.8±3.2)%;主动脉弓部的斑块面积为(64.4±5.4)%,与其他部位的斑块面积相比较,弓部更易发生病变,是其他部位的2~4倍。结论全面改进小鼠主动脉斑块大体油红O染色技术并准确测量小鼠大体及各部位主动脉斑块面积,为评估动脉粥样硬化病变程度提供技术指导。

Abstract:

Objective The gross oil red O staining technique for aortic plaque in mice was improved to accurately evaluate the aortic plaque area in atherosclerotic mice. Methods Aortic dissection was performed on male ApoE-/- mice at the age of 10 months with normal-diet. Microspring scissors were used to remove adventitial fat and to cut longitudinally the entire aorta. The aorta was incubated with oil red O dye in different temperature conditions(25℃, 45℃ and 65℃)for 10 minutes, and then the gross aortawas spread out on the absorbent paper. Photograph was capturedand the plaque area of each part was measured by Image J software. Results Compared with the aortic oil red O staining under the incubation condition at 25℃, the aortic plaques were much more stained than that at 45℃, which showed deep red color and easy for measure. While under the condition of 65℃, plaque shed. Thus, 45℃ was defined as the optimal temperature.Oil red O staining showed significant atherosclerotic plaques in the aortic arch, thoracic aorta, abdominal aorta, and biforked common iliac artery of ApoE-/- miceand the gross aortic plaque size was(29.8±3.2) %. While,the plaque area of the aortic arch was(64.4±5.4) %, whichwas 2-4 times than that of the other parts, indicating that the aortic arch was more prone to lesions than other parts. Conclusion The gross oil red O staining technique for aortic plaque in mice was comprehensively improved and the area of gross and local aortic plaque could be accurately and easily measured, providing a certain technical guidance to evaluate the degree of atherosclerotic lesions.

参考文献

[1] VAN DIEPEN J A,BERBEE J F,HAVEKES L M,et al.Interactions between inflammation and lipid metabolism:Relevance for efficacy of anti-inflammatory drugs in the treatment of atherosclerosis[J].Atherosclerosis,2013,228(2):306-315.

[2] EMINI VESELI B,PERROTTA P,DE MEYER G R A,et al.Animal models of atherosclerosis[J].Eur J Pharmacol,2017,816:3-13.

[3] 王思静,张华,马学宽,等.天香丹对动脉粥样硬化ApoE-/-小鼠IL-6、IL-10表达的影响[J].新疆医科大学学报,2019,42(7):930-933.

[4] SHNERB GANOR R,HARATS D,SCHIBY G,et al.Factor XI deficiency protects against atherogenesis in apolipoprotein E/factor XI double knockout mice[J].Arterioscler Thromb Vasc Biol,2016,36(3):475-481.

[5] LI Y,C G Z,WANG X H,et al.Progression of atherosclerosis in ApoE -knockout mice fed on a high-fat diet[J].Eur Rev Med Pharmacol Sci,2016,20(18):3863-3867.

[6] RIVA G,VILLANOVA M,CIMA L,et al.Oil red O is a useful tool to assess donor liver steatosis on frozen sections during transplantation[J].Transplant Proc,2018,50(10):3539-3543.

[7] ANDRES-MANZANO M J,ANDRES V,DORADO B.Oil red O and hematoxylin and eosin staining for quantification of atherosclerosis burden in mouse aorta and aortic root[J].Methods Mol Biol,2015,1339:85-99.

[8] YAMAMOTO K,MIYOSHI H,CHO K Y,et al.Overexpression of perilipin1 protects against atheroma progression in apolipoprotein E knockout mice[J].Atherosclerosis,2018,269:192-196.

[9] LIN Y,BAI L,CHEN Y,et al.Practical assessment of the quantification of atherosclerotic lesions in ApoE-/- mice[J].Mol Med Rep,2015,12(4):5298-5306.

[10] GOETZ L G,MAMILLAPALLI R,SAHIN C,et al.Addition of estradiol to cross-sex testosterone therapy reduces atherosclerosis plaque formation in female ApoE-/- mice[J].Endocrinology,2018,159(2):754-762.

[11] A P,F B,GD N,et al.The interplay of lipids,lipoproteins,and immunity in atherosclerosis[J].Curr Atheroscler Rep,2018,20(3):12.

[12] KO K A,FUJIWARA K,KRISHNAN S,et al.En face preparation of mouse blood vessels[J].J Vis Exp,2017(123):55460.

[13] HU D,YIN C,MOHANTA S K,et al.Preparation of single cell suspensions from mouse aorta[J].Bio Protoc,2016,6(11):e1832.

[14] 杨美娟,蔡华丹,李健.油红O及苏丹黑B对重度脂肪肝组织脂质染色效果的比较[J].临床与实验病理学杂志,2012,28(02):217-218.

[15] LIU J,LIN J,HE S,et al.Transgenic overexpression of IL-37 protects against atherosclerosis and strengthens plaque stability[J].Cell Physiol Biochem,2018,45(3):1034-1050.

[16] 高红丽,李虹伟,李卫萍,等.吡格列酮对糖尿病ApoE-/-小鼠动脉粥样硬化的影响及机制研究[J].临床和实验医学杂志,2019,18(20):2129-2133.

[17] 王喜欢,张金华,胡亚南,等.大蒜素对高脂饮食ApoE-/-小鼠动脉粥样硬化形成的影响[J].中国动脉硬化杂志,2017,25(02):140-144.

基本信息:

中图分类号:R543.5

引用信息:

[1]陈小翠,马依彤,孙明慧,等.小鼠主动脉斑块大体油红O染色技术的改进[J].新疆医科大学学报,2020,43(10):1308-1311+1317.

基金信息:

新疆维吾尔自治区重点实验室开放课题项目(2017D04001)

投稿时间:

2020-01-10

投稿日期(年):

2020

修回时间:

2020-02-10

终审时间:

2020-10-15

终审日期(年):

2020

审稿周期(年):

1

发布时间:

2020-10-15

出版时间:

2020-10-15

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