硫化氢对脂多糖所致急性肺损伤大鼠血浆和肺组织中炎性细胞因子的影响
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摘要
目的初步探讨硫化氢(H2S)对脂多糖(LPS)所致SD大鼠急性肺损伤(ALI)的保护作用。方法将112只SD大鼠随机分为四组:盐水对照组、内毒素(LPS)组、LPS+(NaHS)组、LPS+炔丙基甘氨酸(PPG)组,每组28只,分别取7只于给药2、4、6、8 h后,应用细胞因子酶联免疫蛋白吸附法(ELISA)法检测各时间点血浆及肺组织中TNF-α、IL-6和IL-4含量,并观察其峰值形成时间变化趋势。结果盐水对照组大鼠的血浆及肺组织中,TNF-α、IL-4浓度均低于检测的下限,IL-6在血浆及肺组织中浓度分别为:(44.03±3.57)ng/L和(48.14±5.68)ng/L;与盐水对照组相比,滴注LPS后大鼠血浆及肺组织中TNF-α、IL-6、IL-4的浓度水平均出现升高,且TNF-α表达峰值的时间也早于IL-6,IL-4的高峰在6 h,要晚于TNF-α、IL-6,但持续时间较长;与LPS组相比,PPG+LPS组大鼠TNF-α、IL-6浓度水平升高,IL-4水平高峰后移至8 h;但NaHS+LPS组大鼠血浆和肺组织中TNF-α、IL-6浓度水平则显著下降,IL-4的水平高峰可前移至4 h(P均<0.01)。结论 H2S能够抑制促炎因子TNF-α、IL-6的增加,并促进抗炎因子IL-4的表达且使其释放高峰提前来减轻机体过度的炎症反应,从而对抗LPS所致急性肺损伤ALI。
Objective To explore the protection effect of hydrogen sulfide(H2S) on endotoxic induced SD rats with acute lung injury(ALI) through replication of endotoxin(LPS) induced ALI model.Methods 112 SD rats were randomly divided into four groups(saline control,LPS,LPS + NaHS,LPS + of PPG groups),28 rats in each group.ELISA assay was used to detect TNF-α,IL-6 and IL-4 content in plasma and lung tissue at each time point after every 7 rats were got according treatment in each group for 2,4,6,8 h respectively and the changes trend of peak formation time was observed.Results In the plasma and lung tissue of rats in control group,TNF-αand IL-4 concentration were below the lower limit of detection levels,IL-6 concentrations in plasma and lung tissue were:(44.03 ± 3.57) ng / L and(48.14 ± 5.68) ng/L respectively.Compared with the control group,after infusion of LPS in rats,the concentration levels of TNF-α,IL-6 and IL-4 in plasma and lung tissue were increased and the TNF-αexpression peak time was earlier than that of IL-6.The IL-4 expression peak time was at 6 h,which was later than that of TNF-α,IL-6 but with a longer duration.Compared with LPS group,the concentration of TNF-αand IL-6 were increased in rats of PPG + LPS group,IL-4 expression peak time moved backward to 8 h.However,the concentration of TNF-α and IL-6 in plasma and lung tissue of rats in NaHS+LPS group were significantly decreased,IL-4 expression peak time moved forward to 4 h(P<0.01 respectively).Conclusions H2S could inhibit the increase of pro-inflammatory cytokines TNF-α,IL-6 and promote anti-inflammatory cytokines IL-4 expression and make its release peak time in advance to reduce the body's excessive inflammatory response,and thus to against LPS induced the ALI.
Objective To explore the protection effect of hydrogen sulfide(H2S) on endotoxic induced SD rats with acute lung injury(ALI) through replication of endotoxin(LPS) induced ALI model.Methods 112 SD rats were randomly divided into four groups(saline control,LPS,LPS + NaHS,LPS + of PPG groups),28 rats in each group.ELISA assay was used to detect TNF-α,IL-6 and IL-4 content in plasma and lung tissue at each time point after every 7 rats were got according treatment in each group for 2,4,6,8 h respectively and the changes trend of peak formation time was observed.Results In the plasma and lung tissue of rats in control group,TNF-αand IL-4 concentration were below the lower limit of detection levels,IL-6 concentrations in plasma and lung tissue were:(44.03 ± 3.57) ng / L and(48.14 ± 5.68) ng/L respectively.Compared with the control group,after infusion of LPS in rats,the concentration levels of TNF-α,IL-6 and IL-4 in plasma and lung tissue were increased and the TNF-αexpression peak time was earlier than that of IL-6.The IL-4 expression peak time was at 6 h,which was later than that of TNF-α,IL-6 but with a longer duration.Compared with LPS group,the concentration of TNF-αand IL-6 were increased in rats of PPG + LPS group,IL-4 expression peak time moved backward to 8 h.However,the concentration of TNF-α and IL-6 in plasma and lung tissue of rats in NaHS+LPS group were significantly decreased,IL-4 expression peak time moved forward to 4 h(P<0.01 respectively).Conclusions H2S could inhibit the increase of pro-inflammatory cytokines TNF-α,IL-6 and promote anti-inflammatory cytokines IL-4 expression and make its release peak time in advance to reduce the body's excessive inflammatory response,and thus to against LPS induced the ALI.
引文
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11 Dahle MK,Overland G,Myhre AE,et al.The phosphatidylinositol 3-kinase/protein kinase B signaling pathway is activated by lipoteichoic acid and plays a role in Kupffer cell production of interleukin-6(IL-6)and IL-10[J].Infect Immun,2004;72(10):5704-11.
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13 Murphey ED,Traber DL.Pretreatment with tumor necrosis factor-alpha attenuates arterial hypotension and mortality induced by endotoxin in pigs[J].Crit Care Med,2000;28(6):2015-21.
14 Cain BS,Meldrum DR,Dinarello CA,et al.Tumor necrosis factor-alpha and interleukin-1 beta synergistically depress human myocardial function[J].Crit Care Med,1999;27(7):1309-18.
2 Downey GP,Dong Q,Kruger J,et al.Regulation of neutruphil activation in Acute lung injury[J].Chest,1999;116(I Supp1):46s-54s.
3 顾俭勇.脂多糖致大鼠急性肺损伤后血浆TNF-α、IL-1β和IL-4的改变[J].山东医药,2009;49(4):41-2.
4 Bone RC.Why sepsis trials fail[J].JAMA,1996;276(7):565-6.
5 Grutz G.New insights into the molecular mechanism of interleukin-10 mediated immunosuppression[J].J Leukoc Biol,2005;77(1):3-15.
6 Marini JJ,Evans TW.Round table conference:acute lung injury[J].Intensive Care Med,1998;24(8):878-83.
7 Dinarello CA.Proinflammatory cytokines[J].Chest,2000;118(6):503-8.
8 刘红菊.肿瘤坏死因子在急性肺损伤发病中的作用[J].国外医学·免疫学分册,1996;5(2):229-31.
9 Amiot F,Fitting C,Tracey KJ,et al.Lipopolysaccharide-induced cytokine cascade and lethality in LT alpha PTNF alpha-deficient mice[J].Mol Med,1997;3(12):864-75.
10 Basak C,Pathak SK,Bhattacharyya A,et al.NF-kappa B-and C/EBP beta-driven interleukin-1βgene expression and PAK1-mediated caspase1 activation play essential roles in interleukin-1βrelease from Helicobacter pylori lipopolysaccharide(LPS)-stimulated macrophages[J].J Biol Chem,2005;280(6):4279-88.
11 Dahle MK,Overland G,Myhre AE,et al.The phosphatidylinositol 3-kinase/protein kinase B signaling pathway is activated by lipoteichoic acid and plays a role in Kupffer cell production of interleukin-6(IL-6)and IL-10[J].Infect Immun,2004;72(10):5704-11.
12 Xie J,Qian J,Wang S,et al.Novel and detrimental effects of lipopolysaccharide on in vitro generation of immature dendritic cells:involvement of mitogen-activated protein kinase p38[J].J Immunol,2003;171(9):4792-800.
13 Murphey ED,Traber DL.Pretreatment with tumor necrosis factor-alpha attenuates arterial hypotension and mortality induced by endotoxin in pigs[J].Crit Care Med,2000;28(6):2015-21.
14 Cain BS,Meldrum DR,Dinarello CA,et al.Tumor necrosis factor-alpha and interleukin-1 beta synergistically depress human myocardial function[J].Crit Care Med,1999;27(7):1309-18.