短期低压低氧对小鼠肠道黏膜免疫功能的影响
|
摘要
目的观测短期低压低氧对小鼠肠道黏膜免疫功能的影响。方法将在海拔2 260 m(青海西宁,大气压582mmHg,PO_2 121.6mmHg,相当于海平面16%的O_2)饲养(IVC)的30只雌性BALB/c小鼠随机分为3组:常氧对照组、低氧7 d组和低氧14 d组。常氧对照组小鼠在西宁饲养14 d,低氧组小鼠分别在模拟5 000 m海拔(大气压405mmHg,PO_2 84.7mmHg,相当于海平面11%的O_2)的低压氧舱饲养7 d和14 d,观察3组小鼠体重、血常规和结肠组织形态学的变化;用酶联免疫吸附(ELISA)法检测肠系膜淋巴结T淋巴细胞培养液上清中白细胞介素-4(IL-4)、白细胞介素10(IL-10)、白细胞介素-17(IL-17)、干扰素-γ(IFN-γ)和肿瘤坏死因子-α(TNF-α)的含量及血清IgG1的含量;同时应用甲苯胺蓝(TB)染色法观察小肠组织肥大细胞的分布及状态。最后对上述结果行相关分析。结果与常氧对照组相比,低氧7 d组血清IgG1含量,小鼠肠系膜淋巴结T淋巴细胞培养液上清中IL-4、IL-10、IFN-γ和IL-17含量均无明显变化,TNF-α含量略有升高,但无统计学意义(P>0.05);结肠组织切片无病理性损伤变化,小肠组织切片中肥大细胞同对照组相比无显著性差异。低氧14 d组血清IgG1含量较低,但无统计学意义(P>0.05);小鼠肠系膜淋巴结T淋巴细胞培养液上清中IL-17的含量明显降低(P<0.05),IL-10和TNF-α的含量明显升高(P<0.05),IL-4和IFN-γ的含量无明显变化;结肠组织切片显示轻微病理性损伤,小肠组织切片显示肥大细胞出现脱颗粒现象。低氧14 d组与7 d组相比,14 d组血清IgG1含量较低,但无统计学意义(P>0.05);小鼠肠系膜淋巴结T淋巴细胞培养液上清中IFN-γ和IL-17的含量明显低于低氧7d组(P<0.05),TNF-α的含量无明显变化,IL-4和IL-10的含量明显升高(P<0.05)。结论短期低压低氧会改变小鼠肠系膜淋巴结T淋巴细胞免疫功能,致Th1细胞/Th2细胞、Th17细胞/Treg细胞亚群的平衡调节紊乱;而对B淋巴细胞参与的体液免疫影响不明显。
Objective To observe the effects of short-term hypobaric hypoxia on the intestinal mucosal immune function in mice.Methods The 30 BALB/c female mice fed at an altitude of 2 260 m(Xining,582 mmHg,PO_2 121.6 mmHg,equivalent to sea level O_2 16%)were randomly divided into 3 groups:normoxia control group,hypoxia 7 d group and hypoxia 14 d group.The normoxic control group was raised in Xining for 14 days.In the hypoxic group,the hypoxic oxygen chamber was simulated at 5 000 m altitude(405 mmHg,PO_2 84.7 mmHg,equivalent to sea level O_2 11%)for 7 days and 14 days.The body weight and blood routine tests of these three groups were observed.The contents in culture supernatant of interleukin-4(IL-4),interleukin-10(IL-10),interleukin-17(IL-17),interferon-gamma(IFN-γ)and tumor necrosis factor-α(TNF-α)were detected in mesenteric lymph node T lymphocytes by ELISA,and IgG1 content of serum by ELISA.HE staining histological changes of colon was observed.The distribution of mast cells were observed by toluidine blue staining in small intestine.Finally,analyzed the above results.Results Compared with the normoxic control group,there was no significant change of IgG1 in serum and there also was no significant change of IL-4,IL-10,IFN-γ and IL-17 in the mouse mesenteric lymph node T cell culture supernatant of HC-7 d group,and the content of TNF-α was slightly increased,but it was no statistical significance(P>0.05).There was no pathological damage in colon tissue sections,and there was no significant difference of the mast cells in the small intestine tissue section.Compared with the control group,there was no significant change of IgG1 in serum in the mouse mesenteric lymph node T cell culture supernatant of HC-14 d group,so it was no statistical significance(P>0.05).The content of IL-17 was significantly decreased(P<0.05)in mesenteric lymph node T cell culture supernatants,IL-10 and TNF-α levels were significantly increased(P<0.05),there was no significant change of IL-4 and IFN-γ.Colon tissue sections have mild pathological damage,degranulation of mast cells in small intestine tissue sections.Compared with HC-7 d group,there was no significant change of IgG1(P>0.05)in the serum of HC-14 d group,the content of IFN-γ and IL-17 was significantly decreased(P<0.05)in the culture supernatant of mouse mesenteric lymph nodes,the content of TNF-α was not significantly changed,and the content of IL-4 and IL-10 was significantly increased(P<0.05)in HC-14 d group.Conclusion The T lymphocyte immunity of mouse mesenteric lymph nodes can be changed in short-term hypobaric hypoxia environment,and the balance of Th1 cells/Th2 cells and Th17 cells/Treg cell subsets will be disordered,it is not obvious effect on humoral immunity of B lymphocytes.
Objective To observe the effects of short-term hypobaric hypoxia on the intestinal mucosal immune function in mice.Methods The 30 BALB/c female mice fed at an altitude of 2 260 m(Xining,582 mmHg,PO_2 121.6 mmHg,equivalent to sea level O_2 16%)were randomly divided into 3 groups:normoxia control group,hypoxia 7 d group and hypoxia 14 d group.The normoxic control group was raised in Xining for 14 days.In the hypoxic group,the hypoxic oxygen chamber was simulated at 5 000 m altitude(405 mmHg,PO_2 84.7 mmHg,equivalent to sea level O_2 11%)for 7 days and 14 days.The body weight and blood routine tests of these three groups were observed.The contents in culture supernatant of interleukin-4(IL-4),interleukin-10(IL-10),interleukin-17(IL-17),interferon-gamma(IFN-γ)and tumor necrosis factor-α(TNF-α)were detected in mesenteric lymph node T lymphocytes by ELISA,and IgG1 content of serum by ELISA.HE staining histological changes of colon was observed.The distribution of mast cells were observed by toluidine blue staining in small intestine.Finally,analyzed the above results.Results Compared with the normoxic control group,there was no significant change of IgG1 in serum and there also was no significant change of IL-4,IL-10,IFN-γ and IL-17 in the mouse mesenteric lymph node T cell culture supernatant of HC-7 d group,and the content of TNF-α was slightly increased,but it was no statistical significance(P>0.05).There was no pathological damage in colon tissue sections,and there was no significant difference of the mast cells in the small intestine tissue section.Compared with the control group,there was no significant change of IgG1 in serum in the mouse mesenteric lymph node T cell culture supernatant of HC-14 d group,so it was no statistical significance(P>0.05).The content of IL-17 was significantly decreased(P<0.05)in mesenteric lymph node T cell culture supernatants,IL-10 and TNF-α levels were significantly increased(P<0.05),there was no significant change of IL-4 and IFN-γ.Colon tissue sections have mild pathological damage,degranulation of mast cells in small intestine tissue sections.Compared with HC-7 d group,there was no significant change of IgG1(P>0.05)in the serum of HC-14 d group,the content of IFN-γ and IL-17 was significantly decreased(P<0.05)in the culture supernatant of mouse mesenteric lymph nodes,the content of TNF-α was not significantly changed,and the content of IL-4 and IL-10 was significantly increased(P<0.05)in HC-14 d group.Conclusion The T lymphocyte immunity of mouse mesenteric lymph nodes can be changed in short-term hypobaric hypoxia environment,and the balance of Th1 cells/Th2 cells and Th17 cells/Treg cell subsets will be disordered,it is not obvious effect on humoral immunity of B lymphocytes.
引文
[1]国际高原医学会慢性高原病专家小组。慢性高原病青海诊断标准[J]。青海医学院学报,2005,26(1):3-5。
[2]汝铁林,郝选明。间歇性低氧训练复合运动训练模型大鼠的造血功能[J]。中国组织工程研究与临床康复,2010,14(37):6913-6916。
[3]张晓娜。高原低氧对小鼠脾脏T淋巴细胞亚群及其功能的影响[D]。青海大学,2016。
[4]Facco M,Zilli C,Siviero M.Modulation of immune response by the acute and chronic exposure to high altitude[J].Med Sci Sports Exerc,2005,37(5):768-774.
[5]Saimm M,Shama SK,Dipti P,et al.Effect of hypobaric hypoxia on immune function in albino rats[J].Int J Biometeorol,1998,42(1):55-59.
[6]Biselli R,Le Moil S,Matricard PM.The effect of hypobaric hypoxia on specific B cell responses following immunization in mice and humans[J].Aviat Space Environ Med,1991,62(9):870-874.
[7]张翠萍,谢印之,张延坤,等。离体淋巴细胞经低氧及低氧预处理后免疫指标的变化[J]。中华航空航天医学杂志,2004,15:129-132。
[8]贾成,郭宜姣,袁东亚。高原环境下汉族人群免疫改变[J]。中华临床免疫和变态反应杂志,2014,8(3):233-237。
[9]Astrid M,Westendorf,Kathrin Skibbe,et al.Hypoxia enhances immunosuppression by inhibiting CD4+ effector T cell function and promoting Treg activity[J].Cellular Physiology and Biochemistry,2017,41:1271-1284.
[10]张晓娜,李积东,胡方杰,等。高原低氧暴露小鼠脾脏T淋巴细胞数量减少且免疫活性下降[J]。细胞与分子免疫学杂志,2017,33(2):164-167。
[11]Tucker A,Mcmurtry IF,Alexander AF,et al.Lung mast cell density and distribution in chronically hypoxic animals[J].J Appl Physiol Respir Environ Exere Physiol,1977,42(2):174-178.
[2]汝铁林,郝选明。间歇性低氧训练复合运动训练模型大鼠的造血功能[J]。中国组织工程研究与临床康复,2010,14(37):6913-6916。
[3]张晓娜。高原低氧对小鼠脾脏T淋巴细胞亚群及其功能的影响[D]。青海大学,2016。
[4]Facco M,Zilli C,Siviero M.Modulation of immune response by the acute and chronic exposure to high altitude[J].Med Sci Sports Exerc,2005,37(5):768-774.
[5]Saimm M,Shama SK,Dipti P,et al.Effect of hypobaric hypoxia on immune function in albino rats[J].Int J Biometeorol,1998,42(1):55-59.
[6]Biselli R,Le Moil S,Matricard PM.The effect of hypobaric hypoxia on specific B cell responses following immunization in mice and humans[J].Aviat Space Environ Med,1991,62(9):870-874.
[7]张翠萍,谢印之,张延坤,等。离体淋巴细胞经低氧及低氧预处理后免疫指标的变化[J]。中华航空航天医学杂志,2004,15:129-132。
[8]贾成,郭宜姣,袁东亚。高原环境下汉族人群免疫改变[J]。中华临床免疫和变态反应杂志,2014,8(3):233-237。
[9]Astrid M,Westendorf,Kathrin Skibbe,et al.Hypoxia enhances immunosuppression by inhibiting CD4+ effector T cell function and promoting Treg activity[J].Cellular Physiology and Biochemistry,2017,41:1271-1284.
[10]张晓娜,李积东,胡方杰,等。高原低氧暴露小鼠脾脏T淋巴细胞数量减少且免疫活性下降[J]。细胞与分子免疫学杂志,2017,33(2):164-167。
[11]Tucker A,Mcmurtry IF,Alexander AF,et al.Lung mast cell density and distribution in chronically hypoxic animals[J].J Appl Physiol Respir Environ Exere Physiol,1977,42(2):174-178.