高脂小鼠脂肪肝组织油红O特殊染色的应用
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摘要
【目的】探讨2种油红O染色法在非酒精性脂肪肝病中脂肪变性检测的应用。【方法】选取非酒精性脂肪肝病动物疾病模型C57BL/6J小鼠30只,安乐死处理后摘取左肝叶,将肝组织用4%多聚甲醛固定液充分固定后分为2份进行组织学制片,1份样品制作石蜡切片,石蜡切片制备完成后进行苏木精-伊红(HE)染色,作为脂肪变性的鉴定,另1份样品用30%蔗糖溶液脱水后用OCT包埋制作冰冻切片,冰冻切片制备完成后进行2种方法的油红O染色。通过盲测的方法,在光学显微镜下观察油红O在脂滴细胞的定位情况,并对2种染色方法进行对比。【结果】2种染色方法对组织及细胞没有任何病理性的损伤,并具有较好的显色与定位作用。【结论】运用良好的特殊染色技术,可以清晰分辨脂滴细胞,并显示出脂滴细胞的数量,从而对非酒精性脂肪肝病中的脂肪变性进行定位与半定量的分析。
[Objective]To explore the application of two oil-red staining methods for steatosis in non-alcoholic fatty liver disease. [Methods] Thirty C57 BL/6 J mice models of non-alcoholic fatty disease were selected. After euthanasia,the left liver lobe was harvested immediately,which was immersed in a large volume of 4% paraformaldehyde fixative in cacodylate buffer. After fixation,the liver was divided into two samples for making histological sections. One sample was made into paraffin sections for hematoxyin-eosin( HE) staining to identify steatosis. The other was made into frozen sections of OCT-embedded block after dehydration with 30% sucrose solution. The frozen sections were stained by two oil-red O staining methods.The localization of oil-red O in the lipid droplet was observed by blind measurement under microscope,and the results obtained by the two staining methods were compared. [Results]The two staining methods proved to be without pathological damage to tissue and cell,and had good coloration and localization effect,which provided scientific basis for the diagnosis of non-alcoholic fatty liver disease. [Conclusion] By the use of good special staining technology,the lipid drop cells are distinguished clearly,and the number of lipid drop cells are displayed. The steatosis in non-alcoholic fatty disease is located and analyzed with half quantification.
[Objective]To explore the application of two oil-red staining methods for steatosis in non-alcoholic fatty liver disease. [Methods] Thirty C57 BL/6 J mice models of non-alcoholic fatty disease were selected. After euthanasia,the left liver lobe was harvested immediately,which was immersed in a large volume of 4% paraformaldehyde fixative in cacodylate buffer. After fixation,the liver was divided into two samples for making histological sections. One sample was made into paraffin sections for hematoxyin-eosin( HE) staining to identify steatosis. The other was made into frozen sections of OCT-embedded block after dehydration with 30% sucrose solution. The frozen sections were stained by two oil-red O staining methods.The localization of oil-red O in the lipid droplet was observed by blind measurement under microscope,and the results obtained by the two staining methods were compared. [Results]The two staining methods proved to be without pathological damage to tissue and cell,and had good coloration and localization effect,which provided scientific basis for the diagnosis of non-alcoholic fatty liver disease. [Conclusion] By the use of good special staining technology,the lipid drop cells are distinguished clearly,and the number of lipid drop cells are displayed. The steatosis in non-alcoholic fatty disease is located and analyzed with half quantification.
引文
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[11]CHU M J,HICKEY A J,TAGALOA S,et al. Ob/ob mouse livers show decreased oxidative phosphorylation efficiencies and anaerobic capacities after cold ischemia[J].PLoS One,2014,9(6):e100609.
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[13]涂琦,刘勇,袁晟.苏木素伊红染色一步法的研究及应用[J].江西医药,2012,47(2):175-176.
[14]全国卫生专业技术资格考试专家委员会. 2015全国卫生专业技术资格考试指导-病理学技术[M].北京:人民卫生出版社,2014:232,438-439.
[15]任秀花,闫爱华,任知春.显示肝、肾上腺脂类油红O方法探讨[J].四川解剖学杂志,2001,9(2):104.
[16]BRUNT E M,JANNEY C G,DI BISCEGLIE A M,et al.Nonalcoholic steatohepatitis:a proposal for grading and staging the histological lesions[J]. Am J Gastroenterol,1999,94(9):2467-2474.
[17]KLEINER D E,BRUNT E M,VAN NATTA M,et al.Design and validation of a histological scoring system for nonalcoholic fatty liver disease[J]. Hepatology,2005,41(6):1313-1321.
[18]王晓颖,王立峰.非酒精性脂肪性肝炎的病理学诊断[J].实用肝脏病杂志,2013,16(6):486-489.
[19]王玉芳,徐晓艳,武彦.常用特殊染色在病理诊断中的应用[J].实用医技杂志,2013,20(12):1357-1358.
[2]PAPPACHAN J M,ANTONIO F A,EDAVALATH M,et al. Non-alcoholic fatty liver disease:a diabetologist's perspective[J]. Endocrine,2014,45(3):344-353.
[3]GOH V J,SILVER D L. The lipid droplet as a potential therapeutic target in NAFLD[J]. Semin Liver Dis,2013,33(4):312-320.
[4]BRUNT E M. Nonalcoholic steatohepatitis:pathologic features and differential diagnosis[J]. Semin Diagn Pathol,2005,22(4):330-338.
[5]黄海燕,辛永宁,姜曼,等.非酒精性脂肪性肝病动物实验模型研究进展[J].临床肝胆病杂志,2014,30(9):948-953.
[6]宣自华,戴敏.高脂诱导非酒精性脂肪肝动物模型及其病理机制研究进展[J].安徽医药,2009,13(3):233-236.
[7]刘静,柳银兰,杨文军,等.高脂高果糖饮食诱导非酒精性脂肪性肝炎小鼠模型的建立和鉴定[J].中华肝脏病杂志,2014,22(6):445-450.
[8]YAMADA H,OHASHI K,SUZUKI K,et al. Longitudinal study of circulating mi R-122 in a rat model of non-alcoholic fatty liver disease[J]. Clinica Chimica Acta,2015,446:267-271.
[9]王肖燕,王金泉,姚刚,等.脂肪组织冰冻切片油红O滴染法的建立[J].家畜生态学报,2014,35(8):58-60,96.
[10]张舵,寨旭,贺西京.油红O染色在大鼠脊髓损伤中的应用[J].中国骨伤,2015,28(8):738-742.
[11]CHU M J,HICKEY A J,TAGALOA S,et al. Ob/ob mouse livers show decreased oxidative phosphorylation efficiencies and anaerobic capacities after cold ischemia[J].PLoS One,2014,9(6):e100609.
[12]KRISTIANSEN M N,VEIDAL S S,RIGBOLT K T,et al.Obese diet-induced mouse models of nonalcoholic steatohepatitis-tracking disease by liver biopsy[J]. World J Hepatol,2016,8(16):673-684.
[13]涂琦,刘勇,袁晟.苏木素伊红染色一步法的研究及应用[J].江西医药,2012,47(2):175-176.
[14]全国卫生专业技术资格考试专家委员会. 2015全国卫生专业技术资格考试指导-病理学技术[M].北京:人民卫生出版社,2014:232,438-439.
[15]任秀花,闫爱华,任知春.显示肝、肾上腺脂类油红O方法探讨[J].四川解剖学杂志,2001,9(2):104.
[16]BRUNT E M,JANNEY C G,DI BISCEGLIE A M,et al.Nonalcoholic steatohepatitis:a proposal for grading and staging the histological lesions[J]. Am J Gastroenterol,1999,94(9):2467-2474.
[17]KLEINER D E,BRUNT E M,VAN NATTA M,et al.Design and validation of a histological scoring system for nonalcoholic fatty liver disease[J]. Hepatology,2005,41(6):1313-1321.
[18]王晓颖,王立峰.非酒精性脂肪性肝炎的病理学诊断[J].实用肝脏病杂志,2013,16(6):486-489.
[19]王玉芳,徐晓艳,武彦.常用特殊染色在病理诊断中的应用[J].实用医技杂志,2013,20(12):1357-1358.