颈动脉体对促炎性细胞因子、慢性缺氧和高压氧刺激的反应
发布时间：2018-08-29 15:53
【摘要】： 哺乳动物的颈动脉体(carotid body, CB)是一个位于颈动脉血管分叉处的血运极其丰富的器官。已经知道,慢性缺氧能引起颈动脉体功能活动增强,而最近的研究表明,颈动脉体支持细胞具有干细胞潜能,缺氧能引起支持细胞表达nestin,而且表达量与缺氧时间呈正相关,但多数临床疾病都是慢性间歇性缺氧,在此状态下nestin和神经再生标记物GAP-43有何变化还未见研究。高压氧作为现代医疗手段中的新成员,对一氧化碳中毒、缺血再灌注等症状具有较好的临床效果,高压氧时颈动脉体的反应如何尚不清楚。PKC作为细胞信号转导的中心环节,在神经传导、递质释放、突触可塑性、长时程增强效应以及肿瘤发生、细胞凋亡等方面起重要作用。ERK是细胞因子、生长因子介导细胞增殖效应中最重要的途径。PKC和ERK信号通路是否在颈动脉体化学感受功能或神经再生过程中发挥作用还有待研究。 白细胞介素-1β(IL-1β)是调节机体免疫、炎症、内分泌和代谢活动急性反应的一种重要的促炎性细胞因子,颈动脉体主细胞本身也表达IL-1β,同时还有其I型受体(IL-1RI)的高表达,提示颈动脉体具有感受促炎性细胞因子刺激的物质基础。促炎性细胞因子可以促进慢性缺氧时颈动脉体中儿茶酚胺类物质的合成,从而调节颈动脉体的慢性缺氧感受功能。既然缺氧能引起支持细胞分化成具有感受功能的主细胞,那么促炎性细胞因子是否也有类似的作用?另外,颈动脉体感受促炎性细胞因子调节的细胞内信号途径也未见报道,对此问题进行研究具有重要理论意义和应用价值。 在本研究中,综合应用免疫组织化学技术、Westren blot技术在组织切片以及蛋白水平,分别研究了慢性低压缺氧、高压氧以及外源性IL-1β刺激对大鼠颈动脉体主细胞TH表达的影响。还初步观察了CB在上述刺激条件下细胞内信号分子pPKC-γ、pERK1/2水平以及神经干细胞和神经再生的标记物Nestin和GAP-43的变化。 结果简要归纳如下: (1)正常条件下,TH、pPKC-γ、pERK1/2、GAP-43、Nestin在大鼠颈动脉体均有表达。TH、pPKC-γ主要在颈动脉体球细胞表达,pERK1/2主要在颈动脉体支持细胞表达,GAP-43在球细胞和支持细胞均有表达。 (2)免疫组织化学结果发现,慢性缺氧和IL-1β刺激都可以引起颈动脉体TH、GAP-43、Nestin表达升高;高压氧刺激抑制TH、GAP-43的表达,高气压刺激无显著差异;慢性缺氧和IL-1β刺激可以引起pERK1/2表达升高,高压氧和高气压刺激也能引起pERK1/2表达升高;慢性缺氧和IL-1β刺激可以引起pPKC-γ表达升高,高压氧刺激则抑制其表达,而高气压刺激则无明显作用。 (3)Western blot进一步证实了上述结果。 主要结论:(1)CB可以感受免疫因子的刺激。(2)缺氧和IL-1β刺激能引起CB生理活性增强,而高压氧则抑制CB生理活性,而且这一效应主要是由于高氧引起的。(3)PKC和ERK信号通路在颈动脉体感受作用中发挥作用。(4)颈动脉体存在分化再生潜能,缺氧和IL-1β刺激能引起CB增殖分化。
[Abstract]:The carotid body (CB) of mammals is an organ with abundant blood supply located at the bifurcation of carotid arteries. Chronic hypoxia has been known to enhance the function of the carotid body. Recent studies have shown that CSCs have stem cell potential, hypoxia can induce nestin expression in CSCs, and hypoxia can induce nestin expression in CSCs. The expression of nestin and GAP-43 is positively correlated with hypoxia time, but most clinical diseases are chronic intermittent hypoxia. There is no study on the changes of nestin and GAP-43 under this condition. Hyperbaric oxygen, as a new member of modern medical methods, has a good clinical effect on carbon monoxide poisoning, ischemia-reperfusion and other symptoms. As the central link of cell signal transduction, PKC plays an important role in nerve conduction, transmitter release, synaptic plasticity, long-term potentiation, tumorigenesis and apoptosis. ERK is the most important pathway of cytokines and growth factors mediating cell proliferation. Whether the signal pathway plays a role in carotid chemosensory function or nerve regeneration is still to be studied.
Interleukin-1 beta (IL-1 beta) is an important pro-inflammatory cytokine that regulates the acute response of immune, inflammatory, endocrine and metabolic activities. The main cell of carotid body itself also expresses IL-1 beta, and has high expression of type I receptor (IL-1RI), suggesting that the carotid body has a material basis for sensing pro-inflammatory cytokine stimulation. Inflammatory cytokines can promote the synthesis of catecholamines in the carotid body during chronic hypoxia, thus regulating the chronic hypoxic sensory function of the carotid body. The intracellular signaling pathways regulated by inflammatory cytokines have not been reported, so it is of great theoretical significance and application value to study this issue.
In this study, the effects of chronic hypobaric hypoxia, hyperbaric oxygen and exogenous IL-1beta stimulation on TH expression in rat carotid somatic host cells were studied by using immunohistochemical technique and Westren blot technique in histological sections and protein levels. The level of K1/2 and the changes of Nestin and GAP-43 in neural stem cells and nerve regeneration markers.
The results are summarized as follows:
(1) Under normal conditions, TH, pPKC-gamma, pERK1/2, GAP-43 and Nestin were expressed in rat carotid body. TH, pPKC-gamma were mainly expressed in carotid body globular cells, pERK1/2 was mainly expressed in carotid body Sertoli cells, and GAP-43 was expressed in globular cells and Sertoli cells.
(2) Immunohistochemical results showed that both chronic hypoxia and IL-1beta stimulation could induce the expression of TH, GAP-43 and Nestin in carotid artery; hyperbaric oxygen stimulation inhibited the expression of TH, GAP-43, while hyperbaric stimulation had no significant difference; chronic hypoxia and IL-1beta stimulation could induce the expression of pERK1/2, hyperbaric oxygen and hyperbaric stimulation could also induce the expression of pERK1/2. Chronic hypoxia and IL-1 beta stimulation could increase the expression of pPKC-gamma, hyperbaric oxygen stimulation could inhibit the expression of pPKC-gamma, but hyperbaric stimulation had no obvious effect.
(3) Western blot further confirmed the above results.
Main conclusions: (1) CB can sense the stimulation of immune factors. (2) Hypoxia and IL-1 beta stimulation can enhance the physiological activity of CB, while hyperbaric oxygen can inhibit the physiological activity of CB, and this effect is mainly due to hyperoxia. (3) PKC and ERK signaling pathways play a role in carotid somatosensory function. (4) There is differentiation and regeneration potential in carotid bodies. Hypoxia and IL-1 beta stimulation can induce proliferation and differentiation of CB.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R363
，
本文编号：2211678
[Abstract]:The carotid body (CB) of mammals is an organ with abundant blood supply located at the bifurcation of carotid arteries. Chronic hypoxia has been known to enhance the function of the carotid body. Recent studies have shown that CSCs have stem cell potential, hypoxia can induce nestin expression in CSCs, and hypoxia can induce nestin expression in CSCs. The expression of nestin and GAP-43 is positively correlated with hypoxia time, but most clinical diseases are chronic intermittent hypoxia. There is no study on the changes of nestin and GAP-43 under this condition. Hyperbaric oxygen, as a new member of modern medical methods, has a good clinical effect on carbon monoxide poisoning, ischemia-reperfusion and other symptoms. As the central link of cell signal transduction, PKC plays an important role in nerve conduction, transmitter release, synaptic plasticity, long-term potentiation, tumorigenesis and apoptosis. ERK is the most important pathway of cytokines and growth factors mediating cell proliferation. Whether the signal pathway plays a role in carotid chemosensory function or nerve regeneration is still to be studied.
Interleukin-1 beta (IL-1 beta) is an important pro-inflammatory cytokine that regulates the acute response of immune, inflammatory, endocrine and metabolic activities. The main cell of carotid body itself also expresses IL-1 beta, and has high expression of type I receptor (IL-1RI), suggesting that the carotid body has a material basis for sensing pro-inflammatory cytokine stimulation. Inflammatory cytokines can promote the synthesis of catecholamines in the carotid body during chronic hypoxia, thus regulating the chronic hypoxic sensory function of the carotid body. The intracellular signaling pathways regulated by inflammatory cytokines have not been reported, so it is of great theoretical significance and application value to study this issue.
In this study, the effects of chronic hypobaric hypoxia, hyperbaric oxygen and exogenous IL-1beta stimulation on TH expression in rat carotid somatic host cells were studied by using immunohistochemical technique and Westren blot technique in histological sections and protein levels. The level of K1/2 and the changes of Nestin and GAP-43 in neural stem cells and nerve regeneration markers.
The results are summarized as follows:
(1) Under normal conditions, TH, pPKC-gamma, pERK1/2, GAP-43 and Nestin were expressed in rat carotid body. TH, pPKC-gamma were mainly expressed in carotid body globular cells, pERK1/2 was mainly expressed in carotid body Sertoli cells, and GAP-43 was expressed in globular cells and Sertoli cells.
(2) Immunohistochemical results showed that both chronic hypoxia and IL-1beta stimulation could induce the expression of TH, GAP-43 and Nestin in carotid artery; hyperbaric oxygen stimulation inhibited the expression of TH, GAP-43, while hyperbaric stimulation had no significant difference; chronic hypoxia and IL-1beta stimulation could induce the expression of pERK1/2, hyperbaric oxygen and hyperbaric stimulation could also induce the expression of pERK1/2. Chronic hypoxia and IL-1 beta stimulation could increase the expression of pPKC-gamma, hyperbaric oxygen stimulation could inhibit the expression of pPKC-gamma, but hyperbaric stimulation had no obvious effect.
(3) Western blot further confirmed the above results.
Main conclusions: (1) CB can sense the stimulation of immune factors. (2) Hypoxia and IL-1 beta stimulation can enhance the physiological activity of CB, while hyperbaric oxygen can inhibit the physiological activity of CB, and this effect is mainly due to hyperoxia. (3) PKC and ERK signaling pathways play a role in carotid somatosensory function. (4) There is differentiation and regeneration potential in carotid bodies. Hypoxia and IL-1 beta stimulation can induce proliferation and differentiation of CB.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R363
，
本文编号：2211678
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