清开灵口服液对内毒素血症模型小鼠心肌炎症因子基因表达及自由基代谢的影响

目的 研究清开灵口服液对内毒素致小鼠心肌细胞受损的保护作用,并从炎症因子表达和自由基代谢方面探讨其作用机制.方法 将48只雄性ICR小鼠随机分为正常组、模型组、清开灵口服液小剂量组、清开灵口服液大剂量组,每组12只.清开灵口服液小、大剂量组分别灌胃清开灵口服液9、18 ml/kg,正常组和模型组灌胃等体积生理盐水,1次/d,共4 d.第5天,除正常组外,其余各组小鼠腹腔注射脂多糖0.2 ml(40 mg/kg)制备内毒素血症模型,正常组注射等体积生理盐水.分别于造模后0.5、8、20 h再次给药.末次给药后1 h,麻醉心脏取血.采用实时荧光定量PCR检测小鼠心肌组织IL-1β、IL-6、TNF-?基因表达,采用ELISA法测定血清IL-1β、IL-6、TNF-?水平及心肌组织MDA、SOD、GSH-Px水平.结果 与模型组比较,清开灵口服液小、大剂量组小鼠血清TNF-?(68.75±7.73)pg/ml、(62.03±16.09)pg/ml比(116.06±21.06)pg/ml]、IL-1β(110.84±40.61)pg/ml、(105.51±38.21)pg/ml比(167.53±54.82)pg/ml]及IL-6(68.78±20.57)pg/ml、(59.71±13.59)pg/ml比(108.80±28.21)pg/ml]水平降低(P<0.01).清开灵口服液小、大剂量组小鼠心肌组织TNF-?mRNA(1.42±0.15)、(1.30±0.46)比(3.00±0.82)],IL-1βmRNA(1.20±0.57)、(1.01±0.40)比(2.32±1.39)]及IL-6 mRNA(1.53±1.10)、(1.16±1.09)比(4.12±2.23)]表达降低(P<0.01或P<0.05).清开灵口服液小、大剂量组小鼠心肌组织MDA(10.64±2.91)nmol/mg、(11.36±3.02)nmol/mg比(15.21±2.31)nmol/mg]降低(P<0.01),SOD(282.32±35.90)U/mg、(325.07±34.76)U/mg比(249.01±45.22)U/mg]和GSH-Px(48.26±17.13)U/g、(49.66±22.11)U/g比(26.47±20.37)U/g]升高(P<0.05).结论 清开灵口服液对内毒素血症导致的心肌损伤具有保护作用,其作用机制可能与下调IL-1β、IL-6、TNF-?基因表达、降低自由基代谢和提高机体抗氧化能力有关.

Effects of Qingkailing on gene expression and free radical metabolism in mice with endotoxemia

Objective To investigate the protective effect of Qingkailing (QKL) in cardiac muscle's injury induced by endotoxin and discuss the mechanism from the inflammatory factors expression and free radical metabolism. Methods A total of 48 male ICR mice were divided into normal group, model group, low dose group and high dose group, 12 mice in each group. The QKL(9 ml/kg) was administered via gavage daily for 4 days in low dose group, the QKL (18 ml/kg) was administered via gavage daily for 4 days in high dose group, the equivalent volume of saline was administered via gavage daily for 4 days in normal group and model group. At the fifth day all groups except normal group, received intraperitoneal injection of LPS 0.2 ml (40 mg/kg), and the normal group received equivalent volume of saline. Intragastric administrated again 0.5h, 8h and 20 h after the model establishment. We took blood from hearts 1 hour after the last administration. The QR-PCR was used to detect the expression of inflammatory factors. The Elisa was used to detect IL-1β, IL-6, TNF-α, MDA, SOD and GSH-Px. Results Compared to the model group, the content of TNF-α (68.75 ± 7.73 pg/ml, 62.03 ± 16.09 pg/ml vs. 116.06 ± 21.06 pg/ml), IL-1β (110.84 ± 40.61 pg/ml, 105.51 ± 38.21 pg/ml vs. 167.53 ± 54.82 pg/ml) and IL-6 (68.78 ± 20.57 pg/ml, 59.71 ± 13.59 pg/ml vs. 108.80 ± 28.21 pg/ml) in low dose group and high dose group were significantly decreased (P<0.01). The expression of TNFα mRNA (1.42 ± 0.15, 1.30 ± 0.46 vs. 3.00 ± 0.82),IL-1β mRNA (1.20 ± 0.57, 1.01 ± 0.40 vs. 2.32 ± 1.39) and IL-6 mRNA (1.53 ± 1.10, 1.16 ± 1.09 vs. 4.12 ± 2.23) in low dose group and high dose group were significantly decreased (P<0.01 or P<0.05). The content of MDA (10.64 ± 2.91 nmol/mg, 11.36 ± 3.02 nmol/mg vs. 15.21 ± 2.31 nmol/mg) in low dose group and high dose group were significantly decreased (P<0.01). The content of SOD (282.32 ± 35.90 U/mg, 325.07 ± 34.76 U/mg vs. 249.01 ± 45.22 U/mg) and GSH-Px (48.26 ± 17.13 U/g, 49.66 ± 22.11 U/g vs. 26.47 ± 20.37 U/g) in low dose group and high dose group were significantly increased (P<0.01). Conclusions The QKL plays a protective role in myocardial injury induced by endotoxemia. Its mechanism may be associated with down-regulation of expression of inflammatory factors, reducing free radicals and improvement of antioxidation.