脂联素对急性呼吸窘迫综合征大鼠炎性因子及细胞凋亡因子的影响

Effect of adiponectin on inflammatory factors and apoptosis factors in rats with ARDS

目的研究脂联素(Adiponectin,APN)对内毒素(Lipopolysaccharide,LPS)诱导的急性呼吸窘迫综合征(ARDS)大鼠炎性因子及细胞凋亡因子的影响。方法采用随机数字表法,将45只健康雄性Wistar大鼠分为三组(每组15只):对照组、LPS组和APN组。对照组和LPS组分别经腹腔注射0.9%氯化钠溶液和LPS(7.5 mg/kg),APN组在造模前12 h经腹腔注射鼠重组APN 6 mg/kg。造模12 h后留取血标本和肺组织标本。采用酶联免疫吸附试验(ELISA)检测各组大鼠血浆及肺组织肿瘤坏死因子α(Tumor necrosis factorα,TNF⁃α)和白细胞介素⁃10(Interleukin 10,IL⁃10)的含量,采用蛋白质印迹法(Western Blot)测定各组大鼠肺组织中的B细胞淋巴瘤/白血病⁃2(Bcl⁃2)、Bcl⁃2相关X蛋白(Bax)、半胱氨酸天冬氨酸蛋白酶3(Caspase⁃3)蛋白相对含量,并对比组间各指标的差异。结果APN组与LPS组的血浆和肺组织中TNF⁃α和IL⁃10水平均高于对照组,且APN组指标低于LPS组[血浆TNF⁃α:9.27(8.55,16.53)ng/L比25.36(22.41,36.34)ng/L,IL⁃10:95.85(77.35,116.01)ng/L比158.51(119.60,346.16)ng/L;肺组织TNF⁃α:9.27(8.55,16.39)ng/L比24.56(18.00,31.11)ng/L;IL⁃10:135.63(127.31,153.21)ng/L比184.94(161.66,203.51)ng/L,均P<0.0167];LPS组和APN组肺组织中Bax、Bcl⁃2、Caspase⁃3蛋白含量均高于对照组,其中APN组的Bcl⁃2高于LPS组[0.41(0.33,0.43)比0.32(0.29,0.36),P<0.0167],而Caspase⁃3低于LPS组[0.23(0.18,0.29)比0.34(0.27,0.45),P<0.0167]。结论APN不仅通过抑制炎症反应降低ARDS大鼠的肺部损伤程度,还可能通过影响组织细胞凋亡来达到肺保护作用。

Objective To investigate the effect of adiponectin(APN)on inflammatory factors and apoptosis factors in rats with Li⁃popolysaccharide(LPS)⁃induced acute respiratory distress syndrome(ARDS).Methods Forty⁃five healthy male Wistar rats were randomly assigned into three groups according to random number table,15 rats in each group,including control group,LPS group and APN group.The control group and LPS group were injected with 0.9%sodium chloride solution and LPS(7.5 mg/kg),respec⁃tively.The APN intervention group was injected with recombinant APN 6 mg/kg through abdominal cavity 12 hours before modeling.Blood samples and lung tissues were collected 12 hours after model establishment.The levels of tumor necrosis factor a(TNF⁃α)and interleukin⁃10(IL⁃10)in plasma and lung tissue in each group were detected by enzyme⁃linked immunosorbent assay(ELI⁃SA).The concentrations of B⁃cell lymphoma/leukemia⁃2(Bcl⁃2),Bcl⁃2 related X protein(Bax),and caspase 3(Caspase⁃3)in lung tissue of each group were determined by Western Blot.The differences of each index were compared in the three groups.Results In APN group and LPS group,the levels of TNF⁃αand IL⁃10 in plasma and lung tissue were higher than those in control group,and the indexes in APN group were lower than those in LPS group[TNF⁃αin plasma:9.27(8.55,16.53)ng/L vs.25.36(22.41,36.34)ng/L;IL⁃10 in plasma:95.85(77.35,116.01)ng/L vs.158.51(119.60,346.16)ng/L;TNF⁃αin lung tissue:9.27(8.55,16.39)ng/L vs.24.56(18.00,31.11)ng/L;IL⁃10 in lung tissue:135.63(127.31,153.21)ng/L vs.184.94(161.66,203.51)ng/L,all P<0.0167].The protein contents of Bax,Bcl⁃2 and Caspase⁃3 in lung tissue of LPS group and APN group were higher than those in control group,and the level of Bcl⁃2 in APN group was higher than that in LPS group[0.41(0.33,0.43)vs.0.32(0.29,0.36),P<0.0167],while Caspase⁃3 was lower than that in LPS group[0.23(0.18,0.29)vs.0.34(0.27,0.45),P<0.0167].Conclusion APN not only reduces the degree of lung injury by inhibiting inflammation,but it may also protect lung tissue by regulating apoptosis.