耐力运动诱导大鼠肾上腺髓质细胞表型转化对EIB的影响
发布时间：2018-07-23 20:14
【摘要】：目的：初步阐明耐力运动在大鼠运动性支气管痉挛发病过程中对肾上腺髓质嗜铬细胞结构和功能的影响及其作用机制。 方法：对大鼠进行中强度或高强度跑台耐力运动,并分别予NGF抗体、力竭运动或卵清蛋白(ovalbumin,OVA)致敏、激发干预,120只雄性Sprague-Dawley大鼠随机分为15组,每组8只,分别为：正常对照组,力竭运动组,中速训练组,中速训练+NGF抗体组,中速训练+力竭运动组,中速训练+NGF抗体+力竭运动组,高速训练组,高速训练+NGF抗体组,高速训练+力竭运动组,高速训练+NGF抗体+力竭运动组,OVA组,OVA+中速训练组,OVA+中速训练+NGF抗体组,OVA+高速训练组,OVA+高速训练+NGF抗体组。检测大鼠气道阻力以及支气管肺泡灌洗液中白细胞比例；ELISA检测大鼠血清中肾上腺素(Epinephrine, EPI)、神经生长因子(Nerve growth factor, NGF)、皮质酮(Corticosterone, CORT)水平；光镜观察大鼠支气管肺组织以及肾上腺髓质的形态学改变；电镜观察大鼠肾上腺髓质嗜铬细胞的超微结构；免疫组化法检测肾上腺髓质中苯乙醇胺-N-甲基转移酶(Phenylethanolamine N-methyl transferase, PNMT)、外周蛋白(Peripherin)的表达；Western blot法检测p-ERK/ERK、p-p38/p38, p-JNK/JNK以及peripherin蛋白表达量；Real-time PCR以及western blot法分别检测JMJD3以及Mash1的mRNA和蛋白表达量。 结果：1.与正常组以及中速训练组比,高速训练组大鼠气道内淋巴细胞浸润明显增加,气道阻力升高；大鼠肾上腺髓质嗜铬细胞出现空泡样变性,嗜铬颗粒减少,出现细胞突起。高速训练组大鼠血清中NGF水平升高,EPI水平下降,肾上腺中PNMT表达下降,peripherin表达上升,同时p-ERK/ERK、JMJD3、Mash1表达均升高；NGF抗体干预能抑制高强度耐力运动所致的上述病理生理改变。 2.与正常组对比,力竭运动组大鼠气道阻力增高,肾上腺髓质中血窦扩张,其血清中EPI、CORT水平升高,肾上腺中PNMT、p-ERK表达增高,而血清中NGF、肾上腺中JMJD3及Mash1表达无明显差异。与高速训练组比较,高速训练+力竭运动组大鼠气道淋巴细胞浸润增加,气道阻力升高,肾上腺中peripherin、p-ERK、JMJD3、Mash1的表达均升高。但血清中EPI、CORT、NGF水平均无明显改变。经NGF抗体干预后,高速训练+NGF抗体+力竭运动组大鼠上述病理生理改变均明显改善。 3.与正常组以及高速训练组比较,OVA组大鼠气道中淋巴细胞、巨噬细胞、嗜酸性粒细胞、中性粒细胞增多,气道阻力明显升高,血清中NGF水平升高,CORT水平下降。与正常对照组比较,OVA组大鼠肾上腺髓质嗜铬颗粒减少,有细胞突起形成,血清EPI水平下降,大鼠肾上腺中PNMT表达下降,peripherin表达上升,同时p-ERK/ERK、JMJD3、Mash1表达均升高。与OVA组对比,OVA+高速训练组大鼠上述病理生理改变进一步加重,嗜铬细胞出现脱颗粒改变,纤维组织增多。而经过NGF抗体以及中强度耐力运动干预,OVA+高速训练+NGF抗体组以及OVA+中速训练组大鼠的病理生理改变可部分逆转。 结论：1.高强度耐力运动能够引起大鼠肾上腺嗜铬细胞向神经细胞表型转化,其发生机制与NGF介导的p-ERK-JMJD3-Mashl通路活化密切关联； 2.剧烈运动激发能够引起高强度耐力运动大鼠支气管痉挛,其机制与肾上腺髓质嗜铬细胞发生神经细胞表型转化导致肾上腺素分泌水平降低密切关联； 3.高强度耐力运动干预与OVA干预相互作用,使大鼠肾上腺髓质嗜铬细胞发生神经细胞表型转化过程加剧,而中强度耐力运动有逆转作用。图38幅,表14个,参考文献67篇
[Abstract]:Objective: to elucidate the effect and mechanism of endurance exercise on the structure and function of adrenal medullary chromaffin cells during the pathogenesis of exercise-induced bronchospasm in rats.
Methods: the rats were treated with moderate or high strength endurance exercise, and were treated with NGF antibody, exhaustive exercise or ovalbumin (OVA) sensitization and excitation intervention. 120 male Sprague-Dawley rats were randomly divided into 15 groups, 8 rats in each group, which were the normal control group, the exhaustive exercise group, the medium speed training group and the medium speed training +NGF antibody group. Speed training + exhausted exercise group, medium speed training +NGF antibody + exhausting exercise group, high speed training group, high speed training +NGF antibody group, high speed training + exhaustive exercise group, high speed training +NGF antibody + exhausted exercise group, OVA group, OVA+ medium speed training group, OVA+ medium speed training +NGF antibody group, OVA+ high-speed training group, OVA+ high-speed training +NGF antibody group. Test rats gas. ELISA, Epinephrine, EPI, Nerve growth factor, NGF, and corticosterone (Corticosterone, CORT) in rat serum, and the morphological changes of bronchopulmonary tissues and adrenal medulla in rats were observed by light microscopy. The rats were observed by electron microscopy. The ultrastructure of the chromaffin cells in the adrenal medulla; the expression of Phenylethanolamine N-methyl transferase (PNMT) (PNMT) and the peripheral protein (Peripherin) in the adrenal medulla by immunohistochemistry; the Western blot method was used to detect p-ERK/ERK, p-p38/p38, p-JNK/JNK, and the expression of peripherin protein. The mRNA and protein expression levels of JMJD3 and Mash1 were detected by Western blot.
Results: 1. compared with the normal group and the medium speed training group, the infiltration of lymphocytes in the airway of the rats in the high speed training group increased obviously, the airway resistance increased, and the chromaffin cells in the adrenal medulla were vacuolated, the chromaffin granules decreased and the cell protruded. The level of NGF in the high speed training group was higher, the level of EPI decreased, and the PN in the adrenal gland was decreased. The expression of MT decreased, the expression of peripherin increased, while the expression of p-ERK/ERK, JMJD3 and Mash1 all increased, and the intervention of NGF antibody could inhibit the above pathophysiological changes caused by high intensity endurance exercise.
2. compared with the normal group, the airway resistance of the rats in the exhaustive exercise group increased and the blood sinus dilated in the adrenal medulla, the level of EPI and CORT in the serum increased, the expression of PNMT and p-ERK in the adrenal gland increased, but there was no significant difference in the expression of JMJD3 and Mash1 in the adrenal gland. Compared with the high speed training group, the high speed training + exhaustive exercise group had lymphatic airway lymphatic. The expression of peripherin, p-ERK, JMJD3, and Mash1 in the adrenal gland increased, but the levels of EPI, CORT and NGF in the serum were not significantly changed. After the prognosis of NGF antibody, the above pathophysiological changes in the rats with high speed training +NGF antibody + exhausting exercise group were obviously improved.
3. compared with the normal group and the high speed training group, the lymphocyte, macrophage, eosinophil, neutrophilic granulocyte, neutrophils increased in the OVA group, increased the airway resistance, the level of NGF in the serum and the level of CORT decreased. Compared with the normal control group, the chromaffin granules in the renal gland medulla of the OVA group were reduced, the formation of cell protruded and the serum EPI was formed. The expression of PNMT in the adrenal gland decreased and the expression of peripherin increased, while the expression of p-ERK/ERK, JMJD3 and Mash1 increased. Compared with the OVA group, the above pathophysiological changes were further aggravated in the OVA+ high speed training group, and the chromaffin cells were degranulation, and the fibrous fabric increased. After NGF antibody and moderate strength endurance exercise intervention, the expression of chromaffin cells was increased. The pathophysiological changes of OVA+ high speed training +NGF antibody group and OVA+ medium speed training group can be partially reversed.
Conclusion: 1. high intensity endurance exercise can induce the phenotype of adrenal chromaffin cells to be transformed into neural cell phenotype, and the mechanism is closely related to the activation of NGF mediated p-ERK-JMJD3-Mashl pathway.
2. intense exercise induced bronchial spasm in rats with high intensity endurance exercise, which is closely related to the reduction of adrenaline secretion by the phenotypic transformation of the adrenal medullary chromaffin cells.
3. the interaction between high strength endurance exercise intervention and OVA intervention makes the phenotypic transformation process of the adrenal medullary chromaffin cells aggravated, while the middle intensity endurance exercise has a reversal effect. Figure 38, table 14, 67 references
【学位授予单位】：中南大学
【学位级别】：博士
【学位授予年份】：2013
【分类号】：R562.25
本文编号：2140520
[Abstract]:Objective: to elucidate the effect and mechanism of endurance exercise on the structure and function of adrenal medullary chromaffin cells during the pathogenesis of exercise-induced bronchospasm in rats.
Methods: the rats were treated with moderate or high strength endurance exercise, and were treated with NGF antibody, exhaustive exercise or ovalbumin (OVA) sensitization and excitation intervention. 120 male Sprague-Dawley rats were randomly divided into 15 groups, 8 rats in each group, which were the normal control group, the exhaustive exercise group, the medium speed training group and the medium speed training +NGF antibody group. Speed training + exhausted exercise group, medium speed training +NGF antibody + exhausting exercise group, high speed training group, high speed training +NGF antibody group, high speed training + exhaustive exercise group, high speed training +NGF antibody + exhausted exercise group, OVA group, OVA+ medium speed training group, OVA+ medium speed training +NGF antibody group, OVA+ high-speed training group, OVA+ high-speed training +NGF antibody group. Test rats gas. ELISA, Epinephrine, EPI, Nerve growth factor, NGF, and corticosterone (Corticosterone, CORT) in rat serum, and the morphological changes of bronchopulmonary tissues and adrenal medulla in rats were observed by light microscopy. The rats were observed by electron microscopy. The ultrastructure of the chromaffin cells in the adrenal medulla; the expression of Phenylethanolamine N-methyl transferase (PNMT) (PNMT) and the peripheral protein (Peripherin) in the adrenal medulla by immunohistochemistry; the Western blot method was used to detect p-ERK/ERK, p-p38/p38, p-JNK/JNK, and the expression of peripherin protein. The mRNA and protein expression levels of JMJD3 and Mash1 were detected by Western blot.
Results: 1. compared with the normal group and the medium speed training group, the infiltration of lymphocytes in the airway of the rats in the high speed training group increased obviously, the airway resistance increased, and the chromaffin cells in the adrenal medulla were vacuolated, the chromaffin granules decreased and the cell protruded. The level of NGF in the high speed training group was higher, the level of EPI decreased, and the PN in the adrenal gland was decreased. The expression of MT decreased, the expression of peripherin increased, while the expression of p-ERK/ERK, JMJD3 and Mash1 all increased, and the intervention of NGF antibody could inhibit the above pathophysiological changes caused by high intensity endurance exercise.
2. compared with the normal group, the airway resistance of the rats in the exhaustive exercise group increased and the blood sinus dilated in the adrenal medulla, the level of EPI and CORT in the serum increased, the expression of PNMT and p-ERK in the adrenal gland increased, but there was no significant difference in the expression of JMJD3 and Mash1 in the adrenal gland. Compared with the high speed training group, the high speed training + exhaustive exercise group had lymphatic airway lymphatic. The expression of peripherin, p-ERK, JMJD3, and Mash1 in the adrenal gland increased, but the levels of EPI, CORT and NGF in the serum were not significantly changed. After the prognosis of NGF antibody, the above pathophysiological changes in the rats with high speed training +NGF antibody + exhausting exercise group were obviously improved.
3. compared with the normal group and the high speed training group, the lymphocyte, macrophage, eosinophil, neutrophilic granulocyte, neutrophils increased in the OVA group, increased the airway resistance, the level of NGF in the serum and the level of CORT decreased. Compared with the normal control group, the chromaffin granules in the renal gland medulla of the OVA group were reduced, the formation of cell protruded and the serum EPI was formed. The expression of PNMT in the adrenal gland decreased and the expression of peripherin increased, while the expression of p-ERK/ERK, JMJD3 and Mash1 increased. Compared with the OVA group, the above pathophysiological changes were further aggravated in the OVA+ high speed training group, and the chromaffin cells were degranulation, and the fibrous fabric increased. After NGF antibody and moderate strength endurance exercise intervention, the expression of chromaffin cells was increased. The pathophysiological changes of OVA+ high speed training +NGF antibody group and OVA+ medium speed training group can be partially reversed.
Conclusion: 1. high intensity endurance exercise can induce the phenotype of adrenal chromaffin cells to be transformed into neural cell phenotype, and the mechanism is closely related to the activation of NGF mediated p-ERK-JMJD3-Mashl pathway.
2. intense exercise induced bronchial spasm in rats with high intensity endurance exercise, which is closely related to the reduction of adrenaline secretion by the phenotypic transformation of the adrenal medullary chromaffin cells.
3. the interaction between high strength endurance exercise intervention and OVA intervention makes the phenotypic transformation process of the adrenal medullary chromaffin cells aggravated, while the middle intensity endurance exercise has a reversal effect. Figure 38, table 14, 67 references
【学位授予单位】：中南大学
【学位级别】：博士
【学位授予年份】：2013
【分类号】：R562.25
【参考文献】
相关期刊论文 前5条
1 李志琴,俞彰,刘懿,沈强,凌诒萍;应急大鼠肾上腺髓质嗜铬细胞胞吐特性的超微结构研究[J];电子显微学报;2003年01期
2 冯俊涛;胡成平;;神经生长因子启动哮喘神经-内分泌-免疫网络功能失衡[J];生物化学与生物物理进展;2008年03期
3 史艳莉;李翠娥;代方梅;黄元汛;;n-3多不饱和脂肪酸对运动性哮喘豚鼠气道炎症和气道高反应性的影响[J];中国运动医学杂志;2010年06期
4 逯勇;黄克武;张洪玉;常晓红;张君;武宝梅;彭晶晶;;冷空气激发前后运动性哮喘患者外周气道功能的变化[J];中国运动医学杂志;2011年01期
5 廖力微,戴路明,黄惠珍,梁咏雪,艾凤英,魏星,陈建华,杨志坚,史有奎;运动性哮喘气道高渗激发后组胺及迷走神经反射的调控作用[J];中华内科杂志;2005年02期
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