合江县斯氏肺吸虫病流行病学调查，JAK-STAT和NF-κβ在此病实验动物肺、肝、脑中的表达与病理观察

  本文关键词：合江县斯氏肺吸虫病流行病学调查，JAK-STAT和NF-κβ在此病实验动物肺、肝、脑中的表达与病理观察，由笔耕文化传播整理发布。

        目的：了解合江县及毗邻地区肺吸虫病流行现状；观察斯氏肺吸虫病实验动物模型肺、肝、脑组织超微病理结构,观察JAK-STAT、NF-Kβ通路表达。方法：溪蟹分部位解剖并计算囊蚴感染率(度),问卷调查,肺吸虫病金标渗滤试剂盒(DIGFA-Kit)检测血清抗体,抗体阳性者做肝功检查；自流行区采集溪蟹,分离斯氏肺吸虫囊蚴,经腹腔注射感染犬,3月后剖杀,取肺、肝组织并检获成虫备用,肺吸虫成虫制备抗原,用脑立体定位仪微量注射肺吸虫抗原至大鼠大脑皮质建立大脑损伤模型,对照组注射生理盐水,24h后解剖大鼠取脑组织；固定、切片。光镜下观察虫体周围和抗原注射周围的组织细胞反应,电镜下观察超微病理结构改变,免疫组织化学方法观察JAK-STAT和NF-Kβ信号通路的表达活性。结果：(1)第二中间宿主以锯齿华溪蟹为主,川南流行区溪蟹的肺吸虫囊蚴携带率为92.37%(2453／2656),虫种鉴定为斯氏肺吸虫(Pagumogonimus Skrjabini),流行区居民食生蟹率21.53%(208／966),皮下游走性包块者4.51%(43／966),流行区人群血清学DIGFA-Kit检测肺吸虫抗体阳性率为4.84%(24／496),川南流行区人群有生食和半生食溪蟹的习惯,且感染与镇村、职业、年龄有关,差异有统计学意义(P<0.05),血清肺吸虫抗体阳性者肝功结果显示Y球蛋白升高,白球蛋白比例倒置；(2)斯氏肺吸虫病肺、肝、脑组织损害的动物模型建立,感染犬肺和肝表面有突起结节,质稍硬,有纤维疤痕形成。HE显示感染犬支气管腔有坏死细胞碎屑或虫卵,肺泡有虫卵沉积,卵周有淋巴细胞浸润,肺泡囊有陷塌,肺泡隔破坏；感染犬肝切片见坏死灶,嗜酸性粒细胞浸润及嗜酸性脓肿形成,坏死周围有肉芽肿及纤维组织包裹,肝细胞气球样变,萎缩、变性、坏死,肝窦扩张；抗原注射大鼠脑血肿周围组织水肿明显,且局灶状坏死,神经元皱缩,尼氏小体溶解,周围出现间隙,神经突呈浅淡或消失,可见单核细胞及胶质细胞反应。电镜显示感染犬肺可见Ⅰ、Ⅱ型上皮细胞肿胀、剥脱,细胞核破坏,线粒体肿胀、嵴断裂,内质网扩张、空泡样变；大量渗出液,嗜酸性粒细胞,单核细胞浸润；肺泡隔破坏,细胞水肿破坏。感染犬肝细胞水肿,细胞器破坏,线粒体肿胀、内质网扩张,肝血窦红细胞破坏,内皮细胞水肿。抗原注射组大鼠大脑皮质神经元胞体肿胀,核移位且形状不规则,核膜凸凹不平,核质溢出；染色质为絮状；线粒体肿胀,粗面内质网扩张呈囊泡化；脂褐素较多。(3)免疫组化显示正常犬肺中STAT和NF-Kβ弱阳性表达；感染犬肺可见STAT和NF-Kβ阳性细胞,免疫阳性物质呈棕褐色,NF-Kβ在肺气道表达多,STAT在肺实质表达多,组间存在显著性差异(X2=476,X2=521,P<0.05)。正常犬肝中STAT和NF-Kβ弱阳性表达：感染犬实验组肝可见STAT和NF-Kβ阳性细胞,以肝细胞坏死区明显,胞质和胞核中可见阳性表达,组间存在显著性差异(X2=405,X2=434,P<0.05)。正常对照组大鼠脑中STAT和NF-Kβ弱阳性表达：抗原注射组大鼠脑中可见STAT和NF-Kβ阳性细胞,以大脑皮质损伤区明显,胞质和胞核中见阳性表达,组间存在显著性差异(X2=476,X2=447,P<0.05)。结论：(1)肺吸虫在肺、肝、脑组织中机械破坏作用和炎症病理反应激活了JAK-STAT、NF-Kβ信号通路的表达。(2)肺吸虫病引起组织超微结构改变：细胞肿胀,线粒体肿胀,内质网扩张。(2)川南流行区肺吸虫病的隐性感染依然存在；肺吸虫病对当地居民尤其是中小学生存在着很大的危害：应加强肺吸虫病防治知识的宣传和教育,制定相应的防制措施,阻断该病在川南地区的流行。

    Study on the epidemic situation of Paragonimiasis Skrjabini in HeJiang country, research on expression of JAK-STAT、and NF-Kβin lung、liver、brain and changes in pathology in experimental paragonimiasisLI Zhen-lian 1,ZHANG Xi-lin 2, WANG Ying2, CHEN Wen-bi1, QIAN Bao-zhen3,, Sugiyama H4, WANG Guang-xi1※(1.Department of Pathogenic Biology,Luzhou Medical College,Luzhou646000, Sichuan Province,China; 2. Department of Pathogenic Biology,Third Military Medical University;3.Insitute of Biological Engineering, Zhejiang Academy of Medical Science; 4. Institutes of National Health in Japan,Japan)Abstract:objective:To survey epidemic situation, clinical manifestation of paragonimiasis skrjabini in southern Sichuan region Hejiang county. To study expression of JAK-STAT, NF-Kβin the lung,liver, brain and changes in ultrastructural pathology in experimental paragonimiasis.Methods:Epidemic situation of Paragonimiasis Skrjabini in HeJiang was surveyed by anatomizing freshwater crab, calculating the rate of infection metacercar-iae, questionaire, detecting serum antibody with DIGFA-kit. Crabs collected from endemic areas, were used to isolate metacercariae of Paragonimus Skrjabini. The dogs were infected with Paragonimus skrjabini metacercariae by intraperitoneal injection, dissected to obtain the lung tissue, liver tissue, and adult worms 3 months after infection. Rat models of brain damage were established by injecting paragonimus adult worm antigen with stereotactic microinjection apparatus,24h later, rat brain tissues were obtained, fixed, sliced. The reaction of cells around the worms and injecting antigen site were observed with the light microscope and the electron microscope, Expression and activity of the JAK-STAT and NF-K 0 signaling pathway were observed by immunohistochemistry. Results:The second intermediate host was Sinopotamon denticulatum. The Paragonimus skrjabini metacercaria infection rate of freshwater crab was 92.37%(2453/2656).The crab species was identified as Pagumogonimus skrjabini.The rate of eating raw crab in residents was 21.53%(208/966), the most remarkable manifestations was migratory subcutaneous nodules (4.51%), The positive rate of antibody of paragonimiasis was 4.84%(24/496).There are tissue damage in experimental dogs and rat, Nodules and formation of fibrous scar on the surface of lung and liver were observed in the infected dog, HE showed necrotic cell debris or eggs in the bronchial lumen and alveolar, lymphocytes infiltration around the egg, alveolar sacs collapsed, alveolar septa damaged. Necrosis of liver, eosinophil infiltration and eosinophilic abscess formation, necrosis and fibrous tissue wrapped around the granuloma, liver cell ballooning degeneration, atrophy, degeneration, necrosis, hepatocyte nuclear enrichment, sinusoidal expansion were observed in experimental dog.Brain edema around the hematoma in the antigen injection rat, focal necrosi, neuron shrinkage, dissolution of Nissl, surrounded by a gap, disappearance of neurites, reaction of mononuclear cells and glial cells were observed in the antigen injection rat.Electron microscopy showed thatⅠ,Ⅱ-type epithelial cell of lung swelling, exfoliation, nuclear damage, mitochondrial swelling, cristae rupture, expansion of endoplasmic reticulum, vacuole, large effusion, eosinophils, mononuclear cell infiltration, alveolar septal damage, edema and destruction of nearly all cells in the infected dog.Liver cell swelling, organelle damage, mitochondrial swelling, endoplasmic reticulum dilation, red blood cell destruction of the liver sinusoids, endothelial cell edema were observed in the infected dog.Cerebral cortex neurons swelling, nuclear translocation and irregular shape, uneven nuclear membrane, nuclear transfer overflow,, flocculent chromatin, mitochondrial swelling, expansion of rough endoplasmic reticulum, more lipofuscin were observed in the antigen injection rat.Immunohistochemistry showed STAT and NF-Kβlow expression in the lung of normal dog, STAT and NF-Kβpositive cells in infected dog, the positive cells showed brown, NF-Kβmore expression in the airway, STAT more expression in the lung parenchyma, significant differences exist between the two groups (X2=476, X2=521, P<0.05).STAT and NF-Kβlow expression in liver of normal dog, visible positive cells of STAT and NF-Kβin the liver of infected dog, significant necrosis of liver cells, expression can be seen in the cytoplasm and nucleus, significant difference exist between the two groups(X2-405, X2= 434, P<0.05). STAT and NF-Kβlow expression in normal rat brain, visible positive cells of STAT and NF-Kβin the brain of antigen injection rat, cortical damage zone significantly, expression can be seen in the cytoplasm and nucleus, significant difference exist between the groups (X2= 476, X2= 447, P<0.05). Conclusion:Mechanical damage and pathological inflammatory response of the lung fluke activate expression of the JAK-STAT, NF-Kβsignaling pathway in lung, liver of infected dogs and brain antigen injection rats. There are paragonimiasis in HeJiang county South Sichuan, To control paragonmiasis in South Sichuan region, comprehensive control measures and health education should be strengthened.

合江县斯氏肺吸虫病流行病学调查，JAK-STAT和NF-κβ在此病实验动物肺、肝、脑中的表达与病理观察

1 合江县斯氏肺吸虫病流行病学调查,JAK-STAT和NF-Kβ在此病实验动物肺、肝、脑中的表达与病理观察3-57    1.1 中文摘要3-6    1.2 英文摘要6-9    1.3 前言10-15    1.4 材料与方法15-22    1.5 结果22-40    1.6 讨论40-48    1.7 结论48-50    1.8 参考文献50-56    1.9 英汉缩略词对照表56-572 致谢57-583 JAK-STAT和NF-Kβ通路在寄生虫病致病机制中研究进展(综述)58-72

  本文关键词：合江县斯氏肺吸虫病流行病学调查，，JAK-STAT和NF-κβ在此病实验动物肺、肝、脑中的表达与病理观察，由笔耕文化传播整理发布。