红芪总多糖对急性肺损伤小鼠细胞因子及抗氧化功能的影响

Influence of total Hedysaripolysaccharide on the level of cytokine and anti-oxidative function in acute lung injury of mice

目的研究红芪总多糖对内毒素(LPS)诱导的急性肺损伤(ALI)的保护作用。方法按照体重将72只小鼠随机分为6组(每组12只):正常组、模型组、对照组及3个剂量实验组。正常组注射等体积量0.9%Na Cl,其余各组腹腔注射10 mg·kg^(-1)LPS,建立小鼠肺损伤模型。在造模后4 h,3个剂量(50,100,200 mg·kg^(-1))实验组每日分别灌服红芪总多糖,对照组每日灌服3 mg·kg^(-1)地塞米松,正常组与模型组每日每只给予0.9%Na Cl,连续3 d。以酶联免疫吸附实验测定肿瘤坏死因子-α(TNF-α)、白细胞介素^(-1)0(IL^(-1)0)表达,用分光光度法检测血清总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)及丙二醛(MDA)含量,以单细胞凝胶电泳检测小鼠淋巴细胞DNA损伤。结果给药后,与模型组小鼠血清TNF-α、IL^(-1)0含量分别为(156.23±7.39),(44.24±2.89)ng·L^(-1)相比,中、大2个剂量实验组的TNF-α、IL^(-1)0含量分别为(139.45±6.57),(35.30±1.88);(120.13±6.81),(33.11±2.71)ng·L^(-1),差异均有统计学意义(均P<0.01)。与模型组小鼠血清T-AOC、SOD水平分别为(5.56±0.13),(67.68±4.86)U·mg^(-1)比较,中、大2个剂量实验组的T-AOC、SOD水平分别为(5.91±0.10),(70.00±3.04);(6.10±0.09),(76.21±4.03)U·mg^(-1),差异均有统计学意义(均P<0.01)。与模型组小鼠血清MDA含量(5.55±0.11)nmol·mg^(-1)相比,中、大2个剂量实验组的MDA含量分别为(5.01±0.11),(4.17±0.12)nmol·mg^(-1),差异均有统计学意义(均P<0.01)。与模型组的小鼠外周血淋巴细胞DNA尾长、尾矩及Olive尾矩分别为(6.45±0.08),(3.22±0.14),(3.35±0.12)μm相比,中、大2个剂量实验组的DNA尾长、尾矩及Olive尾矩分别为(5.81±0.10),(5.24±0.10);(2.86±0.12),(2.71±0.16);(2.67±0.10),(2.17±0.18)μm,差异均有统计学意义(均P<0.01)。结论红芪总多糖对LPS引起的急性肺损伤具有保护作用。其作用机制可能与调节促炎因子TNF-α和抗炎因子IL^(-1)0比例失衡,调节氧化和抗氧化的平衡有关。

Objective To study the protection of total Hedysaripolysaccharide in lipopolysaccharide（ LPS） induced acute lung injury（ ALI） of mice. Methods The mice were randomly divided into 6 groups： normal group,model group（ 8 mg · kg^-1lipopolysaccharide）,control group（ dexamethasone 3 mg·kg^-1）,and experimental groups in low,middle and high dose（ 50,100,200 mg · kg^-1total Hedysaripolysaccharide）.Tumor necrosis factor-α（ TNF-α） and interleukin^-10（ IL^-10）levels in serum were measured with enzyme-linked immunosorbent assay. Total antioxidant capacity（ T-AOC）,the activities of super oxide dismutase（ SOD） and malondialdehyde（ MDA） contents in serum were detected by spectrophotometer. And DNA damage of peripheral blood lymphocytes was observed by single cell gel electrophoresis.Results The serum TNF-α and IL^-10 levels in model group were（ 156. 23 ± 7. 39）,（ 44. 24 ± 2. 89） ng·L^-1,which in middle and high dose of experimental groups were（ 139. 45 ± 6. 57）,（ 35. 30 ± 1. 88）;（ 120. 13 ± 6. 81）,（ 33. 11 ± 2. 71） ng·L^-1,the difference was significant（ all P〈0. 01）. T-AOC,SOD activities in middle and high dose of experimental groups were（ 5. 91 ± 0. 10）,（ 70. 00 ± 3. 04）;（ 6. 10 ± 0. 09）,（ 76. 21 ± 4. 03） U·mg^-1,which were obviously higher than those of model group which were（ 5. 56 ± 0. 13）,（ 67. 68 ± 4. 86） U · mg^-1（ all P〈0. 01）. MDA contents of middle and high dose experimental groups were（ 5. 01 ± 0. 11）,（ 4. 17 ± 0. 12）nmol·mg^-1,which were obviously lower than that of model group with（ 5. 55 ± 0. 11） nmol·mg^-1（ all P〈0. 01）.DNA tail length（ TL） were（ 5. 81 ± 0. 10）,（ 5. 24 ± 0. 10） μm,tail moment（ TM） were（ 2. 86 ± 0. 12）,（ 2. 71 ± 0. 16） μm,and Olive tail moment（ OTM） were（ 2. 67 ± 0. 10）,（ 2. 17 ± 0. 18） μm in middle and high dose of experimental groups,which were significantly lower than those of model group： TL was（ 6. 45 ± 0. 08） μm,TM was（ 3. 22 ± 0. 14） μm,and OTM was（ 3. 35 ± 0. 12） μm,the differences were all significant（ all P〈0. 01）.Conclusion Total Hedysaripolysaccharide had a protective effect against the lipopolysaccharide-induced acute lung injury. The possible mechanisms may be due to the regulation the proportional imbalance of TNF-α and IL^-10,as well as the balance oxidative and anti-oxidative.