摘要
目的探讨血红素氧化酶-1(hemeoxygenase,HO-1)介导脂联素(adiponectin,APN)在脓毒症肺损伤中的保护作用。方法80只雄性Wistar大鼠采用随机数字表法分为4组(每组20只):对照组(C组)、脓毒症模型组(LPS组)、脂联素预处理组(APN组)和HO-1抑制剂锌原卟啉IX(zincprotoporphyrinIX,ZnPPIX)干预组(Zn组)。各组分别于腹腔注射脂多糖(1ipopolysaccharide,LPS)(20rag/ks)或0.9%氯化钠溶液后6h,采用酶联免疫吸附法检测血清中肿瘤坏死因子(tumornecrosisfactor-alpha,TNF咀)和白介素(interlukin-6,IL-6)的含量。采用分光光度法测定一氧化氮(nitricoxide,NO)浓度、肺组织丙二醛(malonaldehyde,MDA)和髓过氧化物酶(myeloperoxidase,MPO)活性变化。采用实时荧光定量聚合酶链反应(real-timePCR)法检测肺组织HO-1、诱导型一氧化氮合成酶(induciblenitricoxidesynthase,iNOS)mRNA的表达水平。观察5d内大鼠生存情况,并绘制生存曲线。结果与c组比较,LPS组、APN组和zn组血清,TNF-α [(321.3±11.2)、(132.5±10.2)、(311.7±12.4)vs(52.1±1.4)ng/L、IL-6[(2229.26±210.25)、(1134.14±11.24)、(2028.27±167.04)vs(55.38±0.23)]ng/L、NO[(103±10)、(79±8)、(97±9)vs(48±3)]]μmol/L和肺组织匀浆MDA[(16.209±0.151)、(5.943±0.310)、(13.238±0.326)vs(3.081±0.017)]]μmol/g、MPO[(12.42±0.46)、(6.77±0.14)、(10.45±0.21)vs(2.74±0.06)]U/g水平显著升高(P〈0.05),生存率显著下降(20%vs 100%;70%vs100%;30%vs100%,P〈0.05);与LPS组比较,APN组血清TNF-α[(132.5±10.2)vs(321.3±11.2)]ng/L、IL-6[(1134.14±107.13)vs(2229.26±210.25)]ng/L、NO[(79±8)vs(103±10)]μmol/L和肺组织匀浆MDA[(5.943±0.310)vs(16.209±0.151)]μmol/g、MPO[(6.77±0,14)vs(12.42±0.46)]U/g水平显著降低(P〈0.05),肺组织HO-1 mRNA[(8.73±0.24)vs(1.54±0.03)]显著升高(P〈0.05),iNOSmRNA[(1.42±0.15)vs(2.01±0.10)]显著下降(P〈0.05),生存率显著增高(70%vs20%,P〈0.05);与APN组比较,Zn组血清TNF-α[(311.7±12.4)vs(132.5±10.2)]ng/L、IL-6[(2028.27±167.04)vs(1134.14±107.13)]ng/L、NO[(97±9)vs(79±8)]μmol/L和肺组织匀浆MDA[(13.238±0.326)vs(5.943±0.310)]μmol/g、MPO[(10.45±0.21)vs(6.77±0.14)]U/g水平显著升高(P〈0.05),肺组织HO-1mRNA[(2.66±0.14)vs(8.73±0.24)]显著下降(P〈0.05),iNOSmRNA[(2.06±0.23)VS(1.42±0.15)]显著升高(P〈0.05),生存率显著下降(30%vs70%,P〈0.05)。结论HO-1可能介导了APN在脓毒症性肺损伤中的保护作用。
Objective To investigate the protective effect of adiponectin (APN) on lung injury in LPS-induced septic rats and whether the protective effect is mediated by heme oxygenase-1 (HO-1). Methods 80 Wistar male rats were randomly divided into 4 groups: control group(C group), LPS group, adiponectin treatment group(APN group) and zinc protoporphyrin IX (ZnPP IX) treatment group (Zn group). At 6h after intraperitoneal injection of lipopolysaccharide (LPS), the serum levels of tumor necrosis factor (TNF)-α and interlukin (IL-6) were detected by ELISA. The serum level of nitric oxide (NO), malonodialdehyde (MDA) content and myeloperoxidase (MPO) activity in lung tissue were detected by spectrophptometry. Lung HO-1,inducible nitric oxide synthase (iNOS) mRNA were detected with real time pohymerase chain reaction (RT-PCR). The mortality rate was calculated 7 d after operation and the survival curves were drawn. Results The serum levels of TNF-α [(321.3±11.2), (132.5±10.2), (311.7± 12.4)vs(52.1±1.4) ] ng/L,IL-6[ (2 229.26±210.25), (1 134.14±11.24), (2 028.27±167.04)vs(55.38±0.23) ] ng/L and NO[ (103± 10), (79±8), (97±9) vs (48±3)] μmol/L in LPS, APN, Zn groups were higher than those in C group (P〈0.05). The MDAconcentration [ ( 16.209±0.151 ), (5.943±0.310), (13.238±0.326) vs (3.081±0.017) ] μmol/g and MPO activity [ (12.42±0.46), (6.77±0. 14), (10.45±0.21 )vs (2.74±0.06)] U/g in lung homogenate in LPS group were higher than those in C group (P〈0.05). The survival rate of rats in LPS,APN,Zn groups were lower than that of C group (20% vs 100% ,70% vs 100%,30% vs 100%, P〈0.05). The serum levels of TNF-α[ (311.7±12.4)vs(132.5±10.2) ] ng/L, IL-6 [ ( 1 134.14±107.13)vs(2 229.26±210.25) ] ng/L and NO[ (79±8) vs(103±10) ] μmol/L in APN group were decreased as eompared with LPS group (P〈0.05). The MDA eaneentration [ (5.943±0.3 10)vs (16.209±0.151) ] μmol/g and MPO aetivity[ (6.77±0.14)vs(12.42±0.46) ] U/g in lung homogenate and the lung iNOS mRNA [ (1.42±0. 15)vs(2.01±0.10)] in APN group were deereased as compared with LPS group (P〈0.05). The lung HO-1 mRNA in APN group was higher than that in LPS group [ (8.73±0.24)vs (1.54±0.03)(P〈0.05 )]. The survival rate of rats in APN group was lower than that of LPS group (70% vs 20%, P〈0.05). The serum levels of TNF-α[ (311.7±12.4)vs(132.5±10.2) ] ng/L, IL-6[ (2 028.27±167.04) vs ( 1 134.14±107.13 )] ng/L and NO [ (97±9)vs(79±8)] μmol/L in Zn group were increased as compared with APN group (P〈0.05). The MDA concentration [(13.238±0.326)vs (5.943±0.310) ]μmol/g and MPO aetivity [ (10.45±0.21)vs (6.77±0.14) ] U/g in lung homogenate and the lung iNOS mRNA[ (2.06±0.23)vs(1.42±0.15) ] in Zn group were raised as eompared with APN group (P〈0.05). The lung HO-1 mRNA [ (2.66±0.14)vs(8.73±0.24)] in group Zn was decreased as compared with APN group(P〈0.05 ). The survival rate of rats in Zn group was lower than that of APN group (30% vs 70%, P〈0.05). Conclusions HO-1 may mediate the proteetive effeet of adiponeetin on lung injury in LPS-indueed septic rats.
出处
《国际麻醉学与复苏杂志》
CAS
2013年第5期390-394,共5页
International Journal of Anesthesiology and Resuscitation