基于肺病模型大鼠病理形态学及相关调控物质变化探讨“肺病及肠”病理传变规律及其机制
发布时间：2018-08-25 16:21
【摘要】：目的：本实验通过建立“肺病”(慢性支气管炎)动物模型,选取三个不同的时间点,观察造模后肺与大肠在病理形态学、细胞组织学、炎症介质等方面的对应性变化,探寻“肺病及肠”的传变规律及其病理传变机制。 方法：选取SD大鼠,雄性,共60只。按体重随机分成空白对照(24只)和模型组(36只)。空白组大鼠置于无烟环境中饲养,模型组大鼠采用单纯烟熏法造模,每次烟熏50min,每天3次(分别在早上9点、下午2点和6点),共烟熏70d。于首次造模第20天、50天、70天随机抽取空白组(8只)和模型组(12只)大鼠检测大鼠肺、胃肠功能；光镜观察肺、胃、十二指肠、空肠、回肠、结肠、直肠组织病理形态学变化；电镜观察肺和结肠组织病理形态学改变；ELISA法检测大鼠肺、结肠组织TNF-α、IL-1β、ET-1、PGE2含量；免疫组化法检测肺、结肠组织VIP、SP、CGRP、iNOS表达。 结果： 1.病理生理 肺功能：与空白组比较,模型组大鼠造模第20天、第50天、第70天呼吸频率增快,潮气量、每分钟通气量降低,有差异(P0.05)或显著性差异(P0.01)；与造模第20天比较,模型组大鼠造模第50天、第70天潮气量、每分钟通气量均降低,有显著性差异(P0.01)。 胃肠功能：与空白组比较,模型组大鼠造模第50天、第70天粪便含水率降低,有显著性差异(P0.01)；第20天、第50天、第70天胃内残留率升高、小肠推进率降低,有显著性差异(P0.01)；与造模第20天比较,模型组大鼠造模第50天、第70天粪便含水率降低、胃内残留率升高、小肠推进率均降低,有差异(P0.05)或显著性差异(P0.01)；与造模第50天比较,模型组大鼠造模第70天粪便含水率降低、胃内残留率升高、小肠推进率降低,有差异(P0.05)或显著性差异(P0.01)。 2.病理形态学 光镜：模型组大鼠造模第20天、第50天、第70天十二指肠、空肠、回肠、直肠、胃组织无明显变化；模型组大鼠肺组织于造模第20天、第50天、第70天均可见支气管广泛上皮细胞变性、坏死,不同程度炎细胞浸润：模型组大鼠结肠组织于造模第20天、第50天、第70天分别有40%、70%、80%大鼠结肠组织出现局部充血水肿,灶性上皮细胞变性、坏死,不同程度的慢性炎细胞浸润,严重者可见小灶性糜烂,黏膜上皮欠完整,腺体排列欠规则,黏膜及黏膜下炎细胞浸润； 电镜：模型组大鼠造模第20天肺组织Ⅰ型肺泡上皮细胞肿胀,线粒体轻度肿胀,Ⅱ型肺泡上皮细胞增生；大鼠结肠组织超微结构病无明显改变。第50天、第70天肺组织Ⅰ型肺泡上皮细胞肿胀,线粒体重度肿胀,Ⅱ型肺泡上皮细胞增生,肺间质内胶原纤维增多、纤维母细胞活跃：大鼠结肠组织出现结肠黏膜上皮表面的微绒毛排列稀疏紊乱,线粒体肿胀,嵴排列紊乱,结肠组织黏膜下固有层间隙胶原纤维增生,见成纤维母细胞。 3.相关调控物质 TNF-α、IL-1β、ET-1、PGE2含量变化：与空白组比较,模型组大鼠造模第20天、第50天、第70天肺组织、结肠组织TNF-α、IL-1β、ET-1、PGE2含量明显升高,有差异(P0.05)或显著性差异(P0.01)；与造模第20天比较,模型组大鼠造模第50天、第70天肺组织、结肠组织TNF-α、IL-1β、ET-1、PGE2含量明显升高,有显著性差异(P0.01)；与造模第50天比较,模型组大鼠造模第70天肺组织、结肠组织TNF-α、IL-1β、ET-1、PGE2含量明显升高,有显著性差异(P0.01)。 VIP、SP、CGRP、iNOS表达变化：与空白组比较,模型组大鼠造模第20天、第50天、第70天肺组织VIP、SP、CGRP、iNOS表达均升高(P0.05或P0.01),第20天结肠组织VIP表达降低,CGRP、iNOS表达升高(P0.05或P0.01),造模第50天、第70天肺组织、结肠组织VIP、SP表达降低,CGRP、iNOS表达升高(P0.05或P0.01)；与造模第20天比较,模型组大鼠造模第50天、第70天肺组织VIP、SP、iNOS表达升高(P0.05或P0.01),模型组大鼠造模第50天结肠组织iNOS表达升高(P0.01),第70天结肠组织VIP、SP、CGRP、iNOS表达升高(P0.05或P0.01)；与造模第50天比较,模型组大鼠造模第70天肺组织VIP、CGRP、iNOS表达升高(P0.05),结肠组织SP、CGRP、iNOS表达升高(PO.05或P0.01)。 结论： 1.肺病大鼠可出现胃肠功能的改变,提示肺病可能传变到“肠”。 2.肺病大鼠可出现大肠的病理变化,提示肺病可能传变到“肠”；肺病大鼠十二指肠、空肠、回肠、直肠、胃组织无明显病理改变,而结肠组织出现不同程度的病理改变,提示肺病传变到“肠”的主要部位可能在结肠。 3.肺病大鼠可出现大肠相关调控物质的变化,提示肺病可能传变到“肠”。 4.初步发现TNF-α、IL-1、ET-1、PGE2等炎症介质可能是“肺病及肠”的物质基础。 5.初步发现VIP、SP、CGRP、iNOS等神经肽物质可能是“肺病及肠”的物质基础。 6.模型大鼠肺病传变到“肠”的时间节点在造模50天左右,肺病是否能传变到“肠”,主要取决于肺脏病变的病理损伤程度,而不单纯取决于造模时间的长短。
[Abstract]:Objective: To observe the corresponding changes of pathological morphology, cytohistology and inflammatory mediators between the lung and the large intestine after the establishment of an animal model of pulmonary disease (chronic bronchitis) and select three different time points.
Methods: Sixty SD rats were randomly divided into two groups according to body weight: control group (24 rats) and model group (36 rats). Rats in blank group were fed in a smoke-free environment. Rats in model group were smoked for 50 minutes three times a day (9:00 a.m., 2:00 p.m. and 6:00 p.m.) for 70 days. The lung, stomach, duodenum, jejunum, ileum, colon and rectum were observed by light microscopy, and the pathological changes of lung and colon were observed by electron microscopy. The contents of TNF-a, IL-1beta, ET-1 and PGE2 in lung and colon were detected by ELISA. The expressions of VIP, SP, CGRP and iNOS in lung and colon tissues were detected by immunohistochemistry.
Result:
1. pathophysiology
Pulmonary function: Compared with the blank group, the respiratory rate increased, tidal volume and minute ventilation volume decreased on the 20th, 50th and 70th day of the model group (P 0.05) or significant difference (P 0.01); compared with the 20th day of the model group, tidal volume and minute ventilation volume decreased on the 50th and 70th day of the model group (P 0.0). 1).
Gastrointestinal function: Compared with the blank group, the water content of feces in the model group decreased on the 50th and 70th day (P Compared with the model group on the fiftieth day, the fecal water content decreased, the gastric residual rate increased, and the intestinal propulsion rate decreased (P 0.05) or significantly (P 0.01).
2. pathomorphology
Light microscopy: There were no significant changes in duodenal, jejunum, ileum, rectum and stomach tissues on the 20th, 50th and 70th day of model rats, and bronchial extensive epithelial cell degeneration, necrosis and inflammatory cell infiltration were observed on the 20th, 50th and 70th day of model rats. On day 50 and day 70, local congestion and edema, focal epithelial degeneration, necrosis, chronic inflammatory cell infiltration were found in 80% of the colonic tissues of rats. In severe cases, focal erosion, incomplete mucosal epithelium, irregular arrangement of glands, and infiltration of mucosal and submucosal inflammatory cells were observed.
Electron microscopy: On the 20th day, the type I alveolar epithelial cells in the lung tissue of the model group were swollen, mitochondria were slightly swollen, and type II alveolar epithelial cells proliferated; the ultrastructural changes of the colon tissue of the rats were not obvious. On the 50th day and the 70th day, the type I alveolar epithelial cells in the lung tissue were swollen, the mitochondria weight was heavily swollen, the type II alveolar epitheli Collagen fibers were increased and fibroblasts were active in the plasma. The microvilli on the surface of colonic mucosa epithelium were arranged sparsely and disorderly. Mitochondria were swollen and cristae were arranged disorderly. Collagen fibers were proliferated and fibroblasts were found in the space between the lamina propria of colonic mucosa.
3. related regulatory substances
Changes of TNF-a, IL-1beta, ET-1 and PGE2 contents: Compared with the blank group, the contents of TNF-a, IL-1beta, ET-1 and PGE2 in lung tissue, colon tissue and colon tissue of the model group increased significantly on the 20th day, 50th day and 70th day (P 0.05) or significant difference (P 0.01); Compared with the 20th day, the contents of TNF-a, IL-1beta, ET-1 and PGE2 in lung tissue and colon tissue of the model group increased significantly (P 0.05) or P 0.01). The contents of TNF-a, IL-1beta, ET-1 and PGE2 in lung tissue, colon tissue and colon tissue were significantly increased on day 70 (P 0.01) compared with day 50 (P 0.01).
Changes of VIP, SP, CGRP, iNOS expression: Compared with the blank group, the expression of VIP, SP, CGRP and iNOS in lung tissue increased on the 20th, 50th and 70th day (P 0.05 or P 0.01), decreased in colon tissue on the 20th day, increased in CGRP and iNOS expression (P 0.05 or P 0.01), decreased in lung tissue and colon tissue on the 50th and 70th day, decreased in CGRP, iNOS expression. The expression of VIP, SP and iNOS in lung tissue increased on the fiftieth and seventieth day (P 0.05 or P 0.01), and the expression of iNOS in colon tissue increased on the fiftieth day (P 0.01), and the expression of VIP, SP, CGRP and iNOS in colon tissue increased on the seventieth day (P 0.05 or P 0.01). The expression of VIP, CGRP and iNOS in lung tissues increased (P 0.05) and the expression of SP, CGRP and iNOS in colon tissues increased (P 0.05 or P 0.01) on the seventieth day.
Conclusion:
1. the change of gastrointestinal function can be seen in rats with lung disease, suggesting that lung disease may change to "intestines".
2. Pathological changes of large intestine may occur in rats with pulmonary disease, suggesting that pulmonary disease may be transmitted to the "intestine"; duodenum, jejunum, ileum, rectum and stomach in rats with pulmonary disease have no obvious pathological changes, but colon tissue has different degrees of pathological changes, suggesting that the main part of pulmonary disease to the "intestine" may be in the colon.
3. the change of large intestine related regulatory substance can be seen in lung disease rats, which suggests that lung disease may change to "intestine".
4. it is preliminarily found that TNF-, IL-1, ET-1, PGE2 and other inflammatory mediators may be the material basis of "lung disease and intestine".
5. preliminary discovery of VIP, SP, CGRP, iNOS and other neuropeptide substances may be the material basis of "lung disease and intestine".
6. Whether the lung disease can be transferred to the intestine or not depends on the degree of pathological damage of the lung lesion, not only on the time of modeling.
【学位授予单位】：成都中医药大学
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R-332
本文编号：2203451
[Abstract]:Objective: To observe the corresponding changes of pathological morphology, cytohistology and inflammatory mediators between the lung and the large intestine after the establishment of an animal model of pulmonary disease (chronic bronchitis) and select three different time points.
Methods: Sixty SD rats were randomly divided into two groups according to body weight: control group (24 rats) and model group (36 rats). Rats in blank group were fed in a smoke-free environment. Rats in model group were smoked for 50 minutes three times a day (9:00 a.m., 2:00 p.m. and 6:00 p.m.) for 70 days. The lung, stomach, duodenum, jejunum, ileum, colon and rectum were observed by light microscopy, and the pathological changes of lung and colon were observed by electron microscopy. The contents of TNF-a, IL-1beta, ET-1 and PGE2 in lung and colon were detected by ELISA. The expressions of VIP, SP, CGRP and iNOS in lung and colon tissues were detected by immunohistochemistry.
Result:
1. pathophysiology
Pulmonary function: Compared with the blank group, the respiratory rate increased, tidal volume and minute ventilation volume decreased on the 20th, 50th and 70th day of the model group (P 0.05) or significant difference (P 0.01); compared with the 20th day of the model group, tidal volume and minute ventilation volume decreased on the 50th and 70th day of the model group (P 0.0). 1).
Gastrointestinal function: Compared with the blank group, the water content of feces in the model group decreased on the 50th and 70th day (P Compared with the model group on the fiftieth day, the fecal water content decreased, the gastric residual rate increased, and the intestinal propulsion rate decreased (P 0.05) or significantly (P 0.01).
2. pathomorphology
Light microscopy: There were no significant changes in duodenal, jejunum, ileum, rectum and stomach tissues on the 20th, 50th and 70th day of model rats, and bronchial extensive epithelial cell degeneration, necrosis and inflammatory cell infiltration were observed on the 20th, 50th and 70th day of model rats. On day 50 and day 70, local congestion and edema, focal epithelial degeneration, necrosis, chronic inflammatory cell infiltration were found in 80% of the colonic tissues of rats. In severe cases, focal erosion, incomplete mucosal epithelium, irregular arrangement of glands, and infiltration of mucosal and submucosal inflammatory cells were observed.
Electron microscopy: On the 20th day, the type I alveolar epithelial cells in the lung tissue of the model group were swollen, mitochondria were slightly swollen, and type II alveolar epithelial cells proliferated; the ultrastructural changes of the colon tissue of the rats were not obvious. On the 50th day and the 70th day, the type I alveolar epithelial cells in the lung tissue were swollen, the mitochondria weight was heavily swollen, the type II alveolar epitheli Collagen fibers were increased and fibroblasts were active in the plasma. The microvilli on the surface of colonic mucosa epithelium were arranged sparsely and disorderly. Mitochondria were swollen and cristae were arranged disorderly. Collagen fibers were proliferated and fibroblasts were found in the space between the lamina propria of colonic mucosa.
3. related regulatory substances
Changes of TNF-a, IL-1beta, ET-1 and PGE2 contents: Compared with the blank group, the contents of TNF-a, IL-1beta, ET-1 and PGE2 in lung tissue, colon tissue and colon tissue of the model group increased significantly on the 20th day, 50th day and 70th day (P 0.05) or significant difference (P 0.01); Compared with the 20th day, the contents of TNF-a, IL-1beta, ET-1 and PGE2 in lung tissue and colon tissue of the model group increased significantly (P 0.05) or P 0.01). The contents of TNF-a, IL-1beta, ET-1 and PGE2 in lung tissue, colon tissue and colon tissue were significantly increased on day 70 (P 0.01) compared with day 50 (P 0.01).
Changes of VIP, SP, CGRP, iNOS expression: Compared with the blank group, the expression of VIP, SP, CGRP and iNOS in lung tissue increased on the 20th, 50th and 70th day (P 0.05 or P 0.01), decreased in colon tissue on the 20th day, increased in CGRP and iNOS expression (P 0.05 or P 0.01), decreased in lung tissue and colon tissue on the 50th and 70th day, decreased in CGRP, iNOS expression. The expression of VIP, SP and iNOS in lung tissue increased on the fiftieth and seventieth day (P 0.05 or P 0.01), and the expression of iNOS in colon tissue increased on the fiftieth day (P 0.01), and the expression of VIP, SP, CGRP and iNOS in colon tissue increased on the seventieth day (P 0.05 or P 0.01). The expression of VIP, CGRP and iNOS in lung tissues increased (P 0.05) and the expression of SP, CGRP and iNOS in colon tissues increased (P 0.05 or P 0.01) on the seventieth day.
Conclusion:
1. the change of gastrointestinal function can be seen in rats with lung disease, suggesting that lung disease may change to "intestines".
2. Pathological changes of large intestine may occur in rats with pulmonary disease, suggesting that pulmonary disease may be transmitted to the "intestine"; duodenum, jejunum, ileum, rectum and stomach in rats with pulmonary disease have no obvious pathological changes, but colon tissue has different degrees of pathological changes, suggesting that the main part of pulmonary disease to the "intestine" may be in the colon.
3. the change of large intestine related regulatory substance can be seen in lung disease rats, which suggests that lung disease may change to "intestine".
4. it is preliminarily found that TNF-, IL-1, ET-1, PGE2 and other inflammatory mediators may be the material basis of "lung disease and intestine".
5. preliminary discovery of VIP, SP, CGRP, iNOS and other neuropeptide substances may be the material basis of "lung disease and intestine".
6. Whether the lung disease can be transferred to the intestine or not depends on the degree of pathological damage of the lung lesion, not only on the time of modeling.
【学位授予单位】：成都中医药大学
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R-332
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