IL-8和Eotaxin在哮喘大鼠中的表达及地塞米松干预研究
发布时间：2018-07-25 16:06
【摘要】：目的:研究哮喘大鼠外周血、肺泡灌洗液(BALF)上清液白介素-8(IL-8)、嗜酸性粒细胞趋化因子(Eotaxin)表达水平和肺组织IL-8、Eotaxin mRNA及蛋白表达及地塞米松(DXM)对其影响。 方法:30只雄性SD大鼠随机分为3组，每组10只：正常组、哮喘组和DXM干预组。（1）哮喘组：第1天和第8天大鼠腹腔注射1ml OVA1mg和Al(OH)3100mg无菌混悬液致敏。2周后将已致敏大鼠置于自制雾化玻璃箱中，，用1%OVA雾化吸入激发，1次/天×30min，哮喘大鼠表现为易激怒、呼吸急促、鼻翼煽动、进食差、大小便失禁等症状，连续7天。（2）DXM干预组于每日激发前半小时予DXM1mg/kg腹腔注射。（3）正常组：致敏与激发均用生理盐水代替。在末次激发后24小时，处死大鼠，收集血清、肺泡灌洗液（BALF）和肺组织。并将肺组织经脱水透明、石蜡包埋等处理后制作成肺标本切片。运用ELISA方法对血清和BALF上清液IL-8、Eotaxin水平进行测定，以及用改良牛氏计数台对BALF沉淀细胞进行计数并将沉淀细胞涂片染色后进行分类计数。观察各组肺组织病理学改变。通过RT-PCR方法测定肺组织IL-8、Eotaxin mRNA表达。运用免疫组织化学方法测定大鼠肺组织标本IL-8、Eotaxin蛋白表达。各组指标数据经统计学方法进行分析比较。 结果:1.哮喘组大鼠BALF中白细胞总数，嗜酸性粒细胞、中性粒细胞、淋巴细胞百分比显著高于正常组(P0.01)；DXM干预组白细胞总数和嗜酸性粒细胞、中性粒细胞、淋巴细胞百分比较哮喘组明显减少(P0.05)，较正常组白细胞总数、嗜酸性粒百分比明显增加(P0.01)，但中性粒细胞、淋巴细胞百分比无统计学意义（P0.05）。 2.哮喘组大鼠血清和BALF上清IL-8、Eotaxin水平均显著高于正常组(P0.01)，DXM干预组IL-8、Eotaxin水平明显低于哮喘组(P0.05)，DXM干预组IL-8水平明显高于正常组(P0.05)，Eotaxin水平与正常组无明显差异（P0.05）。 3.哮喘组大鼠肺组织IL-8、Eotaxin mRNA及蛋白的表达水平明显高于正常组(P0.01)，DXM组IL-8、Eotaxin mRNA及蛋白的表达与哮喘组相比明显减少(P0.01)，仍明显高于正常组(P0.05)。 结论:趋化因子IL-8和Eotaxin参与了哮喘炎症过程，DXM可通过抑制IL-8和Eotaxin表达来减弱其对炎性细胞中性粒细胞和嗜酸性粒细胞的趋化与激活从而减轻炎症反应，这可能是DXM减轻哮喘炎症反应的作用机制之一。
[Abstract]:Aim: to study the expression of interleukin-8 (IL-8) and eosinophil chemokine (Eotaxin) in peripheral blood, alveolar lavage fluid (BALF) supernatant of asthmatic rats and the expression of IL-8 eotaxin mRNA and protein in lung tissue, and the effect of dexamethasone (DXM) on the expression of IL-8 eotaxin and protein. Methods 30 male Sprague-Dawley rats were randomly divided into 3 groups, 10 rats in each group: normal group, Asthma group and DXM intervention group. (1) Asthma group: rats were sensitized by intraperitoneal injection of 1ml OVA1mg and Al (OH) 3100mg aseptic suspension on the 1st and 8th day. The asthmatic rats were irritated by 1%OVA atomization inhalation once a day for 30 minutes. The symptoms of asthma rats were irritable, shortness of breath, agitation of nasal wing, poor food intake, incontinence of feces and urine, and so on. (2) DXM intervention group was injected intraperitoneally with DXM1mg/kg half an hour before daily stimulation. (3) normal group: the sensitization and stimulation were replaced by normal saline. The rats were killed 24 hours after the last stimulation, and the serum, alveolar lavage fluid (BALF) and lung tissue were collected. The lung tissue was treated with dehydration and transparency and paraffin embedded. The levels of IL-8 eotaxin in serum and supernatant of BALF were determined by ELISA method, and the precipitated cells of BALF were counted by modified Bull's counter and stained by smear. Lung histopathological changes were observed in each group. The expression of IL-8 eotaxin mRNA in lung tissue was determined by RT-PCR method. The expression of IL-8 eotaxin in rat lung tissue was determined by immunohistochemical method. The index data of each group were analyzed and compared by statistical method. The result is 1: 1. The percentage of leukocytes, eosinophils, neutrophils and lymphocytes in BALF of asthmatic rats was significantly higher than that of control group (P0.01). The percentage of lymphocytes in asthma group was significantly lower than that in asthma group (P0.05), the percentage of total leukocytes and eosinophilic granulocytes was significantly increased (P0.01), but the percentage of neutrophils and lymphocytes had no statistical significance (P0.05). The level of IL-8 eotaxin in serum and supernatant of BALF in asthma group was significantly higher than that in normal group (P0.01). The level of IL-8 eotaxin in DXM intervention group was significantly lower than that in asthmatic group (P0.05). The level of eotaxin in serum and BALF supernatant of asthmatic group was significantly higher than that in normal group (P0.05). The expression of IL-8 eotaxin mRNA and protein in lung tissue of asthmatic group was significantly higher than that of normal group (P0.01) and the expression of IL-8 eotaxin mRNA and protein in DXM group was significantly lower than that in asthmatic group (P0.01), but still significantly higher than that in normal group (P0.05). Conclusion: the chemokines IL-8 and Eotaxin are involved in the inflammatory process of asthma. DXM can attenuate the chemotaxis and activation of inflammatory neutrophils and eosinophils by inhibiting the expression of IL-8 and Eotaxin. This may be one of the mechanisms by which DXM attenuates the inflammatory response of asthma.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R562.25
本文编号：2144343
[Abstract]:Aim: to study the expression of interleukin-8 (IL-8) and eosinophil chemokine (Eotaxin) in peripheral blood, alveolar lavage fluid (BALF) supernatant of asthmatic rats and the expression of IL-8 eotaxin mRNA and protein in lung tissue, and the effect of dexamethasone (DXM) on the expression of IL-8 eotaxin and protein. Methods 30 male Sprague-Dawley rats were randomly divided into 3 groups, 10 rats in each group: normal group, Asthma group and DXM intervention group. (1) Asthma group: rats were sensitized by intraperitoneal injection of 1ml OVA1mg and Al (OH) 3100mg aseptic suspension on the 1st and 8th day. The asthmatic rats were irritated by 1%OVA atomization inhalation once a day for 30 minutes. The symptoms of asthma rats were irritable, shortness of breath, agitation of nasal wing, poor food intake, incontinence of feces and urine, and so on. (2) DXM intervention group was injected intraperitoneally with DXM1mg/kg half an hour before daily stimulation. (3) normal group: the sensitization and stimulation were replaced by normal saline. The rats were killed 24 hours after the last stimulation, and the serum, alveolar lavage fluid (BALF) and lung tissue were collected. The lung tissue was treated with dehydration and transparency and paraffin embedded. The levels of IL-8 eotaxin in serum and supernatant of BALF were determined by ELISA method, and the precipitated cells of BALF were counted by modified Bull's counter and stained by smear. Lung histopathological changes were observed in each group. The expression of IL-8 eotaxin mRNA in lung tissue was determined by RT-PCR method. The expression of IL-8 eotaxin in rat lung tissue was determined by immunohistochemical method. The index data of each group were analyzed and compared by statistical method. The result is 1: 1. The percentage of leukocytes, eosinophils, neutrophils and lymphocytes in BALF of asthmatic rats was significantly higher than that of control group (P0.01). The percentage of lymphocytes in asthma group was significantly lower than that in asthma group (P0.05), the percentage of total leukocytes and eosinophilic granulocytes was significantly increased (P0.01), but the percentage of neutrophils and lymphocytes had no statistical significance (P0.05). The level of IL-8 eotaxin in serum and supernatant of BALF in asthma group was significantly higher than that in normal group (P0.01). The level of IL-8 eotaxin in DXM intervention group was significantly lower than that in asthmatic group (P0.05). The level of eotaxin in serum and BALF supernatant of asthmatic group was significantly higher than that in normal group (P0.05). The expression of IL-8 eotaxin mRNA and protein in lung tissue of asthmatic group was significantly higher than that of normal group (P0.01) and the expression of IL-8 eotaxin mRNA and protein in DXM group was significantly lower than that in asthmatic group (P0.01), but still significantly higher than that in normal group (P0.05). Conclusion: the chemokines IL-8 and Eotaxin are involved in the inflammatory process of asthma. DXM can attenuate the chemotaxis and activation of inflammatory neutrophils and eosinophils by inhibiting the expression of IL-8 and Eotaxin. This may be one of the mechanisms by which DXM attenuates the inflammatory response of asthma.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R562.25
【参考文献】
相关期刊论文 前2条
1 史菲 ,邱晨;大鼠支气管肺泡灌洗术标准化操作的探讨[J];中国现代医学杂志;2002年24期
2 李昌崇,童夏生,阮正英,李绍波,陈小芳,李孟荣;中性粒细胞弹性蛋白酶和IL-8在大鼠哮喘中的作用及地塞米松调控[J];中华微生物学和免疫学杂志;2005年09期
本文编号：2144343
本文链接：https://www.wllwen.com/yixuelunwen/huxijib/2144343.html
最近更新
教材专著
