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血红素加氧酶-1对哮喘小鼠Treg细胞和Th17细胞平衡的影响 被引量:2

Effect of Heme Oxygenase-1 on the balance of Treg cells and Th17 cells in asthmatic mice
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摘要 目的探讨血红素加氧酶-1(HO-1)对哮喘小鼠Treg细胞和Th17细胞平衡的影响,阐述HO-1的免疫调节作用。方法 60只雌性BALB/c小鼠按数字表法随机分为正常组、哮喘组、高铁氯化血红素(Hemin)组、锡-原卟啉(Sn PP)组、Hemin+si RNA scramble组和Hemin+si RNA HO-1组,每组10只。通过注射和雾化吸入鸡卵蛋白(OVA)制备哮喘模型,正常组以生理盐水代替致敏液。Hemin组和Sn PP组分别腹腔注射Hemin或Sn PP 75μmol/kg,Hemin+si RNA scramble组和Hemin+si RNA HO-1组分别在腹腔注射Hemin前转染p Sico R-GFP-si RNA scramble或p Sico R-GFP-si RNA HO-1。末次激发后24 h内,Western blot和比色法分别检测各组小鼠肺部组织中HO-1蛋白表达和HO-1活性;HE染色观察肺部组织病理变化;流式细胞术检测外周血中Treg细胞、Th17细胞占CD4^+T细胞的百分比,ELISA法检测血清和支气管肺泡灌洗液(BALF)中IL-10、IL-17的水平。结果与正常组相比,哮喘组小鼠肺部HO-1表达和活性有所增加,炎性反应明显,Treg/CD4^+T细胞的百分比和IL-10水平显著降低,Th17/CD4^+T细胞的百分比和IL-17水平显著升高,差异具有统计学意义(P<0.05)。与哮喘组相比,Hemin组明显增加HO-1表达和活性,减少肺部组织炎性反应,增加Treg/CD4^+T细胞的百分比和IL-10水平,降低Th17/CD4^+T细胞的百分比和IL-17水平,而Sn PP组没有明显变化。与Hemin组相比,Hemin+si RNA HOv1组肺部组织中HO-1表达和活性降低,炎性反应增加,Treg/CD4^+T细胞的百分比和IL-10水平降低,Th17/CD4^+T细胞的百分比和IL-17水平增加,差异具有统计学意义(P<0.05)。结论增加HO-1的表达和活性能够明显改善哮喘小鼠炎性反应,增加Treg细胞的数目并促进IL-10的产生,同时抑制Th17细胞的表达及IL-17的产生。 Objective To investigate the effect of Heme Oxygenase-1(HO-1) on the balance of Treg cells and Th17 cells in asthmatic mice, and to explain the immunomodulatory effect of HO-1. Methods Sixty female BALB/c mice were randomly divided into six groups according to random number table: normal control group, asthma group, Hemin group, tin- protoporphyrin(Sn PP) group, Hemin+si RNA scramble group and Hemin+si RNA HO-1 group, with 10 mice in each group. Asthmatic mouse model was established by intraperitoneal injection and aerosol inhalation of ovalbumin(OVA), while mice in the normal control group were given normal saline instead. Hemin group and Sn PP group were intraperitoneally injected with Hemin or 75 μ mol/kg Sn PP, respectively. Hemin + si RNA scramble group and Hemin+si RNA HO-1 group were transfected with p Sico R-GFP-si RNA scramble or p Sico R-GFP-si RNA HO-1 before intraperitoneal injection with Hemin. All mice were sacrificed at 24 hours after the last excitation. Western blot and HO-1 activity quantitative Kit were respectively used to measure the expression and activity of HO-1 in lung tissues. The hematoxylin-eosin staining was used to observe the airway structural changes. Flow cytometry was used to measure the percentages of Treg cells and T helper(Th17) cells in CD4~+T cells in peripheral blood, while enzyme-linked immunosorbent assay(ELISA) was used to measure the levels of interleukin(IL)-10 and IL-17 in serum and bronchoalveolar lavage fluid. Results Compared with the normal control group, asthma group showed an increase in the expression and activity of HO-1, and inflammatory reaction was obvious. The percentage of Treg/CD4~+T cells and the level of IL-10 were significantly decreased, and percentage of Th17/CD4~+T cells and the level of IL-17 were significantly increased,with statistically significant differences(P0.05). Compared with the asthma group, Hemin group had significantly increased expression and activity of HO-1, inhibited inflammatory reaction in lung tissue, increased percentage of Treg cells and I-10 level, decreased percentage of Th17 cells and IL-17 level. However, there was no significant change in the Sn PP group. Compared with the Hemin group, Hemin +si RNA HO-1 group showed an obvious decrease in the ex-pression and activity of HO-1, percentage of Treg cells and IL-10 level, and an increase in inflammatory reaction, percentage of Th17 cells, and IL-17 level. The differences were statistically significant(P0.05). Conclusion Increasing the expression and activity of HO-1 could significantly improve the inflammatory reaction, increase the number of Treg cells and promote the production of IL-10, and meanwhile inhibit the generation of Th17 cells and production of IL-17 in asthmatic mice.
作者 杨欣 张庆 张艳萍 魏晓晔 姬亚梅 YANG Xin ZHANG Qing ZHANG Yan-ping WEI Xiao-ye JI Ya-mei(Department of Paediatrics, YuLin Children's Hospital, Yulin 719000, Shaanxi, CHINA Department of Paediatrics, Xingyuan Hosptial of Yulin, Yulin 719000, Shaanxi, CHINA)
出处 《海南医学》 CAS 2017年第1期9-14,共6页 Hainan Medical Journal
关键词 血红素加氧酶-1 哮喘 CD4+CD25+调节性T细胞 Th17细胞 Heme oxygenase-l Asthma CD4+CD25+ regulatory T cell T helper cell(Th17) cell
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