摘要
目的分离银星竹鼠巨噬细胞并鉴定,观察不同组织部位来源巨噬细胞对马尔尼菲青霉分生孢子的吞噬作用并测定其吞噬能力,为进一步探讨竹鼠与马尔尼菲青霉及马尔尼菲青霉病的相互关系奠定基础。方法使用不同方法分别从银星竹鼠肺泡、腹腔、脾脏中分离获得巨噬细胞。采用瑞氏-吉姆萨染色、透射电镜观察细胞形态结构;透射电镜观察不同组织来源巨噬细胞对马尔尼菲青霉分生孢子的吞噬作用及吞噬前后细胞内外结构的变化情况;采用流式细胞技术检测不同组织来源巨噬细胞对马尔尼菲青霉分生孢子的吞噬率。结果成功分离获得不同组织来源的竹鼠原代巨噬细胞,通过光镜、电镜观察证实所分离细胞均具有巨噬细胞的形态结构特征;所分离巨噬细胞在吞噬马尔尼菲青霉分生孢子后,细胞表面伪足增多增粗、吞噬空泡增多,线粒体、内质网、溶酶体等细胞器未见明显的数量及结构的改变,被巨噬细胞吞噬的马尔尼菲青霉分生孢子也未见明显结构变化;流式细胞技术检测肺泡、腹腔、脾脏巨噬细胞对马尔尼菲青霉分生孢子的吞噬率分别为75.09%±6.92%、83.36%±0.87%、78.84%±3.73%,统计学分析三者间存在统计学差异(P<0.05)。结论成功分离获得不同组织来源的竹鼠巨噬细胞,不同组织来源巨噬细胞对马尔尼菲青霉分生孢子具有吞噬作用,且均有较高吞噬率;透射电镜下未见竹鼠巨噬细胞对马尔尼菲青霉分生孢子明显的杀伤表现,可能两者间存在一种共生关系。
The objective of this study was to isolate macrophages from Rhizomys pruinosus,and provide methods for preliminary identification of macrophages based on morphological and phagocytic characteristics.Meanwhile,this research also aimed at observing the phagocytosis of different sources of macrophages to Penicillium marneffei conidia and detecting the phagocytic rate,which will provide initial experimental evidence of the correlation among Penicillium marneffei,Penicilliosis marneffei and Rhizomyidae.Different methods were used to isolate macrophages from alveoli,peritoneum and spleen of Rhizomys pruinosus.The cell morphology was observed by Wright-Giemsa staining and transmission electron microscopy.By transmission electron microscopy,the phagocytosis of different sources of macrophages to Penicillium marneffei conidia and the changes of the surface and intracellular structure of macrophages before and after phagocytosis were observed.The phagocytic rate was detected by flow cytometry.Results displayed that different sources of primary macrophages of Rhizomys pruinosus were isolated.The isolated cells had the characteristic of macrophages in morphology which have been confirmed by light microscope and electron microscope.At the same time,the number of cell surface projection increased and became thicker.However,the number and structure of mitochondria,endocytoplasmic reticulum and lysosome did not change after the phagocytosis.The structure of Penicillium marneffei conidia had no change,either.The phagocytic rates of alveolar,peritoneal and spleen macrophages to Penicillium marneffei conidia were 75.09%±6.92%,83.36%±0.87%,and 78.84%±3.73%,respectively,with statistically significant difference among these three groups（P0.05）.It＇s implied that the macrophages isolated from alveoli,peritoneum and spleen of Rhizomys pruinosus have good phagocytosis to Penicillium marneffei conidia.No lethal effect of the Rhizomys pruinosus macrophages on Penicillium marneffei conidia was observed by transmission electron microscopy.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2012年第1期51-55,共5页
Chinese Journal of Zoonoses
基金
NSFC-广东联合基金重点项目(U0932003)
关键词
银星竹鼠
巨噬细胞
马尔尼菲青霉
分生孢子
吞噬
Rhizomys pruinosus
macrophage
Penicillium marneffei
conidia
phagocytosis
