【摘要】：目的：本實(shí)驗(yàn)通過建立“肺病”(慢性支氣管炎)動(dòng)物模型,選取三個(gè)不同的時(shí)間點(diǎn),觀察造模后肺與大腸在病理形態(tài)學(xué)、細(xì)胞組織學(xué)、炎癥介質(zhì)等方面的對(duì)應(yīng)性變化,探尋“肺病及腸”的傳變規(guī)律及其病理傳變機(jī)制。 方法：選取SD大鼠,雄性,共60只。按體重隨機(jī)分成空白對(duì)照(24只)和模型組(36只)�？瞻捉M大鼠置于無煙環(huán)境中飼養(yǎng),模型組大鼠采用單純煙熏法造模,每次煙熏50min,每天3次(分別在早上9點(diǎn)、下午2點(diǎn)和6點(diǎn)),共煙熏70d。于首次造模第20天、50天、70天隨機(jī)抽取空白組(8只)和模型組(12只)大鼠檢測(cè)大鼠肺、胃腸功能；光鏡觀察肺、胃、十二指腸、空腸、回腸、結(jié)腸、直腸組織病理形態(tài)學(xué)變化；電鏡觀察肺和結(jié)腸組織病理形態(tài)學(xué)改變；ELISA法檢測(cè)大鼠肺、結(jié)腸組織TNF-α、IL-1β、ET-1、PGE2含量；免疫組化法檢測(cè)肺、結(jié)腸組織VIP、SP、CGRP、iNOS表達(dá)。 結(jié)果： 1.病理生理 肺功能：與空白組比較,模型組大鼠造模第20天、第50天、第70天呼吸頻率增快,潮氣量、每分鐘通氣量降低,有差異(P0.05)或顯著性差異(P0.01)；與造模第20天比較,模型組大鼠造模第50天、第70天潮氣量、每分鐘通氣量均降低,有顯著性差異(P0.01)。 胃腸功能：與空白組比較,模型組大鼠造模第50天、第70天糞便含水率降低,有顯著性差異(P0.01)；第20天、第50天、第70天胃內(nèi)殘留率升高、小腸推進(jìn)率降低,有顯著性差異(P0.01)；與造模第20天比較,模型組大鼠造模第50天、第70天糞便含水率降低、胃內(nèi)殘留率升高、小腸推進(jìn)率均降低,有差異(P0.05)或顯著性差異(P0.01)；與造模第50天比較,模型組大鼠造模第70天糞便含水率降低、胃內(nèi)殘留率升高、小腸推進(jìn)率降低,有差異(P0.05)或顯著性差異(P0.01)。 2.病理形態(tài)學(xué) 光鏡：模型組大鼠造模第20天、第50天、第70天十二指腸、空腸、回腸、直腸、胃組織無明顯變化；模型組大鼠肺組織于造模第20天、第50天、第70天均可見支氣管廣泛上皮細(xì)胞變性、壞死,不同程度炎細(xì)胞浸潤：模型組大鼠結(jié)腸組織于造模第20天、第50天、第70天分別有40%、70%、80%大鼠結(jié)腸組織出現(xiàn)局部充血水腫,灶性上皮細(xì)胞變性、壞死,不同程度的慢性炎細(xì)胞浸潤,嚴(yán)重者可見小灶性糜爛,黏膜上皮欠完整,腺體排列欠規(guī)則,黏膜及黏膜下炎細(xì)胞浸潤； 電鏡：模型組大鼠造模第20天肺組織Ⅰ型肺泡上皮細(xì)胞腫脹,線粒體輕度腫脹,Ⅱ型肺泡上皮細(xì)胞增生；大鼠結(jié)腸組織超微結(jié)構(gòu)病無明顯改變。第50天、第70天肺組織Ⅰ型肺泡上皮細(xì)胞腫脹,線粒體重度腫脹,Ⅱ型肺泡上皮細(xì)胞增生,肺間質(zhì)內(nèi)膠原纖維增多、纖維母細(xì)胞活躍：大鼠結(jié)腸組織出現(xiàn)結(jié)腸黏膜上皮表面的微絨毛排列稀疏紊亂,線粒體腫脹,嵴排列紊亂,結(jié)腸組織黏膜下固有層間隙膠原纖維增生,見成纖維母細(xì)胞。 3.相關(guān)調(diào)控物質(zhì) TNF-α、IL-1β、ET-1、PGE2含量變化：與空白組比較,模型組大鼠造模第20天、第50天、第70天肺組織、結(jié)腸組織TNF-α、IL-1β、ET-1、PGE2含量明顯升高,有差異(P0.05)或顯著性差異(P0.01)；與造模第20天比較,模型組大鼠造模第50天、第70天肺組織、結(jié)腸組織TNF-α、IL-1β、ET-1、PGE2含量明顯升高,有顯著性差異(P0.01)；與造模第50天比較,模型組大鼠造模第70天肺組織、結(jié)腸組織TNF-α、IL-1β、ET-1、PGE2含量明顯升高,有顯著性差異(P0.01)。 VIP、SP、CGRP、iNOS表達(dá)變化：與空白組比較,模型組大鼠造模第20天、第50天、第70天肺組織VIP、SP、CGRP、iNOS表達(dá)均升高(P0.05或P0.01),第20天結(jié)腸組織VIP表達(dá)降低,CGRP、iNOS表達(dá)升高(P0.05或P0.01),造模第50天、第70天肺組織、結(jié)腸組織VIP、SP表達(dá)降低,CGRP、iNOS表達(dá)升高(P0.05或P0.01)；與造模第20天比較,模型組大鼠造模第50天、第70天肺組織VIP、SP、iNOS表達(dá)升高(P0.05或P0.01),模型組大鼠造模第50天結(jié)腸組織iNOS表達(dá)升高(P0.01),第70天結(jié)腸組織VIP、SP、CGRP、iNOS表達(dá)升高(P0.05或P0.01)；與造模第50天比較,模型組大鼠造模第70天肺組織VIP、CGRP、iNOS表達(dá)升高(P0.05),結(jié)腸組織SP、CGRP、iNOS表達(dá)升高(PO.05或P0.01)。 結(jié)論： 1.肺病大鼠可出現(xiàn)胃腸功能的改變,提示肺病可能傳變到“腸”。 2.肺病大鼠可出現(xiàn)大腸的病理變化,提示肺病可能傳變到“腸”；肺病大鼠十二指腸、空腸、回腸、直腸、胃組織無明顯病理改變,而結(jié)腸組織出現(xiàn)不同程度的病理改變,提示肺病傳變到“腸”的主要部位可能在結(jié)腸。 3.肺病大鼠可出現(xiàn)大腸相關(guān)調(diào)控物質(zhì)的變化,提示肺病可能傳變到“腸”。 4.初步發(fā)現(xiàn)TNF-α、IL-1、ET-1、PGE2等炎癥介質(zhì)可能是“肺病及腸”的物質(zhì)基礎(chǔ)。 5.初步發(fā)現(xiàn)VIP、SP、CGRP、iNOS等神經(jīng)肽物質(zhì)可能是“肺病及腸”的物質(zhì)基礎(chǔ)。 6.模型大鼠肺病傳變到“腸”的時(shí)間節(jié)點(diǎn)在造模50天左右,肺病是否能傳變到“腸”,主要取決于肺臟病變的病理損傷程度,而不單純?nèi)Q于造模時(shí)間的長(zhǎng)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.
【學(xué)位授予單位】：成都中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2012
【分類號(hào)】：R-332

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