【摘要】： 目的觀察Wistar大鼠肺組織中卡氏肺孢子蟲的生長發(fā)育和形態(tài)結(jié)構(gòu)。 方法通過皮下注射地塞米松磷酸鈉建立Wistar大鼠卡氏肺孢子蟲肺炎動物模型為實(shí)驗(yàn)組,另設(shè)不注射藥物為對照組。制備大鼠肺印片標(biāo)本,瑞-姬氏染色,光學(xué)顯微鏡下檢測和觀察卡氏肺孢子蟲。取大鼠肺組織標(biāo)本,制備肺組織切片和透射、掃描電鏡觀察標(biāo)本,分別使用光鏡、透射和掃描電鏡觀察該蟲生長發(fā)育、形態(tài)結(jié)構(gòu)及其感染大鼠肺病理學(xué)變化。用數(shù)字圖像分析技術(shù),對卡氏肺孢子蟲包囊進(jìn)行形態(tài)學(xué)參數(shù)測定。 結(jié)果肺印片標(biāo)本經(jīng)瑞-姬氏染色,光學(xué)顯微鏡油鏡下檢測,可見清晰卡氏肺孢子蟲包囊和滋養(yǎng)體,以肺泡內(nèi)多見,偶可在細(xì)胞內(nèi)見到。對照組大鼠未檢測到該蟲。實(shí)驗(yàn)組大鼠肺印片標(biāo)本該蟲檢出率為57%,第4～5周查到包囊為主,7～8周后以滋養(yǎng)體為主。掃描電鏡觀察,卡氏肺孢子蟲有表面光滑和粗糙兩種類型,其中粗糙型又有微絨毛、管狀結(jié)構(gòu)、顆粒樣物三種情況;觀察到壁上有凹陷、正在脫囊及伸出偽足的蟲體。透射電鏡觀察到卡氏肺孢子蟲有滋養(yǎng)體、囊前期和包囊三個發(fā)育階段。滋養(yǎng)體多附著于肺Ⅰ型上皮細(xì)胞,偶見附著于肺Ⅱ型上皮細(xì)胞,有的伸出1或多個較寬大偽足,部分細(xì)胞質(zhì)內(nèi)發(fā)現(xiàn)核相關(guān)細(xì)胞器和紡錘微管。囊前期有早、中、晚三種類型,在其內(nèi)發(fā)現(xiàn)有代表減數(shù)分裂的聯(lián)會復(fù)合體。包囊壁上有一增厚處,內(nèi)有一孔隙。第10周實(shí)驗(yàn)組大鼠肺間隔及氣管中可見到卡氏肺孢子蟲。用數(shù)字圖像處理技術(shù)測量卡氏肺孢子蟲包囊長徑、短徑、投影面積、體積、周長、等效直徑和圓球度數(shù)值均數(shù)分別為:4.8159μm,3.6923μm,13.07μm~2,36.76μm~3,14.29μm,4.03μm和0.8。光鏡觀察HE染色的肺組織切片,第7～8周實(shí)驗(yàn)組大鼠肺泡內(nèi)被大量蟲體和滲出物充滿,肺Ⅱ型上皮細(xì)胞脫落,肺泡隔增厚。掃描電鏡觀察肺內(nèi)有卡氏肺孢子蟲,肺組織呈纖維性改變。透射電鏡觀察,肺Ⅱ型上皮細(xì)胞質(zhì)內(nèi)可見許多空泡和電子密度高的顆粒。含蟲體的肺Ⅱ型上皮細(xì)胞表膜突出,形成質(zhì)膜小泡,細(xì)胞核膜內(nèi)陷,核內(nèi)染色體呈絲狀并繞成團(tuán)。 結(jié)論瑞-姬氏復(fù)染法檢測卡氏肺孢子蟲,操作簡單、蟲體清晰,有利于一般實(shí)驗(yàn)室對該蟲檢測。掃描電鏡觀察到表面有粗細(xì)不等的管狀物和正在脫囊的蟲體,陽性大鼠肺組織纖維化改變,為國內(nèi)首次報道。透射電鏡觀察到卡氏肺孢子蟲的滋養(yǎng)體、囊前期和包囊三個發(fā)育階段;滋養(yǎng)體附著于肺Ⅱ型上皮細(xì)胞及氣管內(nèi),國內(nèi)外未見報道;包囊壁上觀察到一增厚處,內(nèi)有一孔隙,為罕見報道。數(shù)字圖像分析技術(shù)獲得了卡氏肺孢子蟲包囊的一系列數(shù)據(jù),豐富了肺孢子蟲生物學(xué)資料,為病原學(xué)診斷提供參考資料。
[Abstract]:Objective to observe the growth, development and morphology of Pneumocystis carinii in lung tissue of Wistar rats. Methods the animal model of pneumocystis carinii pneumonia was established by subcutaneous injection of dexamethasone sodium phosphate in Wistar rats. Rat lung impressions were prepared and stained by Riggi's staining. Pneumocystis carinii was detected and observed under optical microscope. Lung tissue sections and transmission were prepared from rat lung tissue. Light microscope, transmission electron microscope and scanning electron microscope were used to observe the growth and development, morphological structure and pathological changes of lung of infected rats. The morphological parameters of the cysts of Pneumocystis carinii were measured by digital image analysis. Results the pulmonary specimens were stained by Ricky's stain and detected by oil microscope. The clear cysts and trophozoites of Pneumocystis carinii were found in the alveoli and occasionally in the cells. The worm was not detected in the control group. In the experimental group, the positive rate of the worm was 57%, the cyst was found in the 4th week, and the trophozoite was the main one after 78 weeks. Scanning electron microscope (SEM) showed that Pneumocystis carinii had smooth surface and rough surface, including rough type and microvilli, tubular structure and granulocytes, and there were indentation in the wall, exfoliation and extension of pseudopodia on the wall of Pneumocystis carinii. Transmission electron microscopy showed that Pneumocystis carinii had trophozoites, precysts and cysts. Most trophozoites were attached to type I epithelial cells, and occasionally to type 鈪，

本文編號：2316317