黑斑蛙外周血细胞的细胞化学染色特征和细胞化学谱  

Cytochemical staining features and cytochemical profiles of peripheral blood cells in Rana nigromaculata

作　　者：王琴 李莉 苏洁洁 陈娴娴 胡雨佳 张盛周[1] Wang Qin;Li li;Su Jiejie;Chen Xianxian;Hu Yujia;Zhang Shengzhou(College of Life Science,Anhui Normal University,Key Laboratory for Conservation and Use of Important Biological Resources of Anhui Province,Wuhu 241000,China)

机构地区：[1]安徽师范大学生命科学学院,安徽省重要生物资源保护与利用重点实验室,芜湖241000

出　　处：《中国组织化学与细胞化学杂志》2024年第5期429-435,共7页Chinese Journal of Histochemistry and Cytochemistry

基　　金：安徽省自然科学基金项目(1808085MC82);安徽省重要生物资源保护与利用重点实验室建设基金。

摘　　要：目的研究黑斑蛙(Rana nigromaculata)外周血细胞的细胞化学染色特征及细胞化学谱。方法于黑斑蛙心腔抽取血液样本,存放于EDTA抗凝管中,随后制作血涂片,血涂片按照一定次序进行瑞氏及7种细胞化学染色,主要包括酸性磷酸酶染色(ACP)、碱性磷酸酶染色(AKP)、氯乙酸AS-D萘酚酯酶染色(AS-D)、α-醋酸萘酚酯酶染色(α-NAE)、过碘酸-雪夫染色(PAS)、过氧化物酶染色(POX)和苏丹黑B染色(SBB)。结果在黑斑蛙外周血中观察到红细胞、5种类型的白细胞和血栓细胞,5种白细胞即淋巴细胞、单核细胞、嗜中性粒细胞、嗜酸性粒细胞和嗜碱性粒细胞。红细胞PAS染色呈阳性,ACP、AKP、AS-D、α-NAE、POX和SBB染色均为阴性;淋巴细胞ACP和α-NAE染色为阳性,AS-D和PAS染色为弱阳性,AKP、POX和SBB染色为阴性;单核细胞POX染色呈强阳性,ACP和AS-D染色呈阳性,α-NAE、PAS和SBB染色为弱阳性,AKP染色为阴性;嗜中性粒细胞AS-D、POX和SBB染色为阳性,ACP、α-NAE和PAS染色为弱阳性,AKP染色呈阴性;嗜酸性粒细胞POX染色呈强阳性,α-NAE和SBB染色为阳性,ACP、AS-D和PAS染色为弱阳性,AKP染色呈阴性;嗜碱性粒细胞POX染色呈强阳性,AS-D和PAS染色为阳性,ACP和α-NAE染色为弱阳性,AKP和SBB染色为阴性;血栓细胞α-NAE染色呈阳性,ACP和PAS染色为弱阳性,AKP、AS-D、POX和SBB染色均为阴性。结论黑斑蛙外周血细胞的细胞化学谱和其他两栖类有相同之处,也有一些明显的物种特异性。本研究结果可为黑斑蛙人工养殖中健康监测和疾病诊断提供参考资料。Objective To study the cytochemical staining characteristics and cytochemical profile of peripheral blood cells in the black-spotted frog(Rana nigromaculata).Methods Blood samples were collected from the heart cavity of Rana nigromaculata and stored in EDTA anticoagulant tubes.Blood smears were then prepared and subjected to a series of cytochemical stains,including Wright’s stain and seven specific cytochemical stains:acid phosphatase(ACP),alkaline phosphatase(AKP),chloracetate esterase(AS-D),α-naphthyl acetate esterase(α-NAE),periodic acid-Schiff(PAS),peroxidase(POX),and Sudan black B(SBB).Results Peripheral blood of the Rana nigromaculata contained erythrocytes,five types of leukocytes,and thrombocytes.The five types of leukocytes were lymphocytes,monocytes,neutrophils,eosinophils,and basophils.Erythrocytes were positive for PAS staining,while ACP,AKP,AS-D,α-NAE,POX,and SBB were all negative.Lymphocytes showed positive staining for ACP and α-NAE,weak positive for AS-D and PAS,and negative for AKP,POX,and SBB.Monocytes showed strong positive staining for POX,positive for ACP and AS-D,weak positive forα-NAE,PAS,and SBB,and negative for AKP.Neutrophils were positive for AS-D,POX,and SBB,weak positive for ACP,α-NAE,and PAS,and negative for AKP.Eosinophils showed strong positive staining for POX,positive forα-NAE and SBB,weak positive for ACP,AS-D,and PAS,and negative for AKP.Basophils exhibited strong positive staining for POX,positive for AS-D and PAS,weak positive for ACP and α-NAE,and negative for AKP and SBB.Thrombocytes were positive for α-NAE,weak positive for ACP and PAS,and negative for AKP,AS-D,POX,and SBB.Conclusion The cytochemical profile of peripheral blood cells in the Rana nigromaculata shares similarities with other amphibians,but also displays some species-specific characteristics.The results of this study provide valuable reference data for health monitoring and disease diagnosis in black-spotted frog farming.

关 键 词：黑斑蛙 外周血 血细胞 细胞化学 

分 类 号：Q954.6[生物学—动物学]