硫化氫在臭氧誘導(dǎo)的小鼠氧化應(yīng)激模型中的作用機(jī)制
發(fā)布時(shí)間：2018-08-05 16:39
【摘要】：背景與目的急性臭氧暴露可導(dǎo)致氣道炎癥,氣道上皮細(xì)胞損傷,氧化應(yīng)激和氣道高反應(yīng)性(Airway hyperresponsiveness,AHR)。硫化氫(Hydrogen sulfide,H2S)作為新發(fā)現(xiàn)的氣體信號(hào)分子,在炎癥反應(yīng)和氧化應(yīng)激反應(yīng)中具有重要作用。內(nèi)源性H2S主要由胱硫醚-β-合成酶(Cystathionine-β-synthase,CBS)和胱硫醚-γ-裂解酶(Cystathionine-γ-yase,CSE)催化產(chǎn)生。本研究的目的是探討H2S能否預(yù)防急性臭氧暴露的危害。研究方法小鼠隨機(jī)分為空氣暴露組和臭氧暴露組。臭氧暴露濃度為3ppm,持續(xù)3小時(shí)。每次暴露前給予小鼠腹腔注射硫氫化鈉(Na HS,H2S供體)或炔丙基甘氨酸(PPG,H2S生成酶CSE抑制劑)。對(duì)照組給予磷酸鹽緩沖液(PBS)。暴露結(jié)束24小時(shí)后進(jìn)行研究。在另一個(gè)體外實(shí)驗(yàn)中,通過膜片鉗技術(shù)觀察Na HS對(duì)小鼠氣道平滑肌細(xì)胞大電導(dǎo)鈣激活鉀通道(Large-conductance calcium activated K+channels,BKCa)的作用。結(jié)果急性臭氧暴露下調(diào)H2S/CSE、CBS體系,降低血清H2S水平和CBS、CSE m RNA及蛋白的表達(dá)。給予外源性H2S供體Na HS可抑制臭氧誘導(dǎo)的AHR和氣道炎癥,降低氧化應(yīng)激水平,減少氣道上皮細(xì)胞凋亡,抑制H2S/CSE、CBS體系的下調(diào)。不僅如此,Na HS可抑制臭氧誘導(dǎo)的p38絲裂原活化蛋白激酶(p38 MAPK)和熱休克蛋白27(HSP27)的磷酸化,但Na HS可促進(jìn)Akt(亦被稱為蛋白激酶B)的磷酸化。給予H2S生成酶CSE抑制劑PPG將減少內(nèi)源性H2S水平,并導(dǎo)致臭氧對(duì)肺組織的損傷進(jìn)一步惡化。體外實(shí)驗(yàn)顯示Na HS可增加小鼠氣道平滑肌細(xì)胞Ca2+依賴性外向鉀電流。結(jié)論本研究結(jié)果表明,H2S在臭氧誘導(dǎo)的氧化應(yīng)激模型中具有保護(hù)作用。這種保護(hù)作用與H2S抑制p38 MAPK和HSP27的磷酸化,以及激活A(yù)kt的磷酸化有關(guān)。此外,H2S可激活小鼠氣道平滑肌細(xì)胞的BKCa通道。H2S可望成為氣道炎癥性疾病新的治療手段。
[Abstract]:Background & objective Acute ozone exposure can lead to airway inflammation, airway epithelial cell injury, oxidative stress and Airway hyperresponsiveness (AHR). As a newly discovered gas signal molecule, hydrogen sulfide (Hydrogen sulfide H2S) plays an important role in inflammatory reaction and oxidative stress reaction. Endogenous H 2S is mainly produced by Cystathionine- 尾 -synthase (CBS) and cystathionine- 緯 -lyase (CSE). The purpose of this study was to investigate whether H 2S can prevent acute ozone exposure. Methods mice were randomly divided into air exposure group and ozone exposure group. The ozone exposure concentration was 3 ppm and lasted for 3 hours. Mice were given intraperitoneal injection of sodium sulphide (NaHSH _ 2S donor) or propargyl glycine (CSE inhibitor) before each exposure. The control group was given phosphate buffer solution (PBS). The study was conducted 24 hours after exposure. In another experiment in vitro, the effects of NaHS on Large-conductance calcium activated K channels activated potassium channels (BK Ca) in mouse airway smooth muscle cells were observed by patch clamp technique. Results Acute ozone exposure down-regulated the CBS system of H _ 2S / CSE, decreased serum H _ 2S level and the expression of CSE m RNA and protein in serum. Exogenous H 2S donor NaHS inhibited ozone induced AHR and airway inflammation, decreased oxidative stress, reduced airway epithelial cell apoptosis, and inhibited the downregulation of H 2S / CSE / CBS system. Moreover, NaHS inhibited the phosphorylation of p38 mitogen-activated protein kinase (p38 MAPK) and heat shock protein 27 (HSP27) induced by ozone, but NaHS promoted the phosphorylation of Akt (also known as protein kinase B). Administration of CSE inhibitor of H2S-producing enzyme PPG could reduce endogenous H2S level and lead to further deterioration of ozone damage to lung tissue. In vitro experiments showed that NaHS could increase Ca2 dependent outward potassium currents in mouse airway smooth muscle cells. Conclusion the results suggest that H2S has protective effect on oxidative stress induced by ozone. This protective effect is related to the inhibition of phosphorylation of p38 MAPK and HSP27 and activation of phosphorylation of Akt by H 2S. In addition, H2S can activate the BKCa channel of mouse airway smooth muscle cells. H2S may be a new therapy for airway inflammatory diseases.
【學(xué)位授予單位】：上海交通大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R-332;R56
本文編號(hào)：2166365
[Abstract]:Background & objective Acute ozone exposure can lead to airway inflammation, airway epithelial cell injury, oxidative stress and Airway hyperresponsiveness (AHR). As a newly discovered gas signal molecule, hydrogen sulfide (Hydrogen sulfide H2S) plays an important role in inflammatory reaction and oxidative stress reaction. Endogenous H 2S is mainly produced by Cystathionine- 尾 -synthase (CBS) and cystathionine- 緯 -lyase (CSE). The purpose of this study was to investigate whether H 2S can prevent acute ozone exposure. Methods mice were randomly divided into air exposure group and ozone exposure group. The ozone exposure concentration was 3 ppm and lasted for 3 hours. Mice were given intraperitoneal injection of sodium sulphide (NaHSH _ 2S donor) or propargyl glycine (CSE inhibitor) before each exposure. The control group was given phosphate buffer solution (PBS). The study was conducted 24 hours after exposure. In another experiment in vitro, the effects of NaHS on Large-conductance calcium activated K channels activated potassium channels (BK Ca) in mouse airway smooth muscle cells were observed by patch clamp technique. Results Acute ozone exposure down-regulated the CBS system of H _ 2S / CSE, decreased serum H _ 2S level and the expression of CSE m RNA and protein in serum. Exogenous H 2S donor NaHS inhibited ozone induced AHR and airway inflammation, decreased oxidative stress, reduced airway epithelial cell apoptosis, and inhibited the downregulation of H 2S / CSE / CBS system. Moreover, NaHS inhibited the phosphorylation of p38 mitogen-activated protein kinase (p38 MAPK) and heat shock protein 27 (HSP27) induced by ozone, but NaHS promoted the phosphorylation of Akt (also known as protein kinase B). Administration of CSE inhibitor of H2S-producing enzyme PPG could reduce endogenous H2S level and lead to further deterioration of ozone damage to lung tissue. In vitro experiments showed that NaHS could increase Ca2 dependent outward potassium currents in mouse airway smooth muscle cells. Conclusion the results suggest that H2S has protective effect on oxidative stress induced by ozone. This protective effect is related to the inhibition of phosphorylation of p38 MAPK and HSP27 and activation of phosphorylation of Akt by H 2S. In addition, H2S can activate the BKCa channel of mouse airway smooth muscle cells. H2S may be a new therapy for airway inflammatory diseases.
【學(xué)位授予單位】：上海交通大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R-332;R56
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