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凉膈散对内毒素致急性肺损伤大鼠高迁移率族蛋白1影响

周勇1 赵玮2

310003 杭州市红十字会医院干部保健科1;310006 杭州,浙江中医药大学附属第一医院呼吸生理研究中心2

周勇,Email: 13216701315@163.com

浙江省中医药管理局课题(2011ZA083)

摘 要:目的 观察凉膈散对内毒素脂多糖(LPS)致急性肺损伤(ALI)大鼠高迁移率族蛋白1(HMGB1)表达的影响,探讨凉膈散治疗ALI的抗炎机制。方法 选取健康雄性SD大鼠72只,随机分为6组,每组12只,分别为正常对照组、ALI模型组、凉膈散大剂量组、凉膈散中剂量组、凉膈散小剂量组和泼尼松治疗组。给予正常对照组大鼠气管内滴注生理盐水,其余各组大鼠均气管内滴注LPS,每天一次,连续2 d,气道内滴注1 h后分别以生理盐水、凉膈散不同剂量、泼尼松灌胃,每天2次。分别于第二次气管滴注后12 h、24 h每组处死6只。取肺组织,采用荧光定量PCR和Western blot检测HMGB1基因和蛋白的表达,各组指标的均数比较均采用SPSS 19.0软件进行单因素方差分析,P<0.05为差异具有统计学意义。结果 12 h肺组织HMGB1 mRNA灰度值模型组(1.365 5±0.449 2)高于正常对照组(0.389 8±0.169 6)(P<0.01),泼尼松组(0.600 8±0.212 8)、凉膈散大剂量组(0.670 7±0.217 5)、中剂量组(0.731 5±0.200 1)、小剂量组(0.799 3±0.552 6)均低于模型组(1.365 5±0.449 2)(P<0.01),HMGB1相对光密度值模型组(30.03±0.15)高于正常对照组(12.20±0.10)(P<0.01),泼尼松组(13.47±0.25)、凉膈散大剂量组(15.70±0.20)、中剂量组(18.60±0.20)、小剂量组(22.47±0.25)均低于模型组(30.03±0.15)(P<0.01);24 h肺组织HMGB1 mRNA灰度值模型组(1.710 9±0.802 8)高于正常对照组(0.691 8±0.245 3)(P<0.01),泼尼松组(0.727 0±0.097 7)、凉膈散大剂量组(0.782 5±0.178 0)、中剂量组(0.943 3±0.113 6)、小剂量组(1.125 5±0.213 7)均低于模型组(1.710 9±0.802 8)(P<0.01或P<0.05),HMGB1相对光密度值模型组(30.63±0.25)高于正常对照组(10.27±0.31)(P<0.01),泼尼松组(13.63± 0.15)、凉膈散大剂量组(18.63±0.21)、中剂量组(22.57±0.21)、小剂量组(24.53±0.32)均低于模型组(30.03±0.15)(P<0.01或P<0.05)。结论 凉膈散可抑制肺组织HMGB1蛋白及mRNA表达,从而减轻ALI肺部炎症反应。

关键词:急性肺损伤; 凉膈散; 高迁移率族蛋白质1; 高迁移率族蛋白1 mRNA

Effect of Lianggesan on high mobility group protein B1 (HMGB1) in a rat model of lipopolysaccharide-induced acute lung injury

Zhou Yong1, Zhao Wei2.

Department of Leader Health, Hangzhou Red Cross Hospital, Hangzhou 310003, China; 2Research Center of Respiration Physiological Functions, the First Affiliated Hospital of Zhejiang University of Chine

Zhou Yong, Email: 13216701315@163.com

Abstract:Objective To investigate the effect of Lianggesan on the expression of HMGB1 and discuss the antiinflammatory mechanism of Lianggesan in ALI. Methods 72 healthy male SD rats were randomly divided into 6 groups (n=12): normal control group, model group, the prednisone group and the Lianggesan decoction high, middle and low dose groups. The LPS solution (2 mg/kg) was instilled into the trachea to prepare rat ALI model except normal control group, once a day for two days. Each treatment group completed the second infusion 1 hour after LPS solution. The drug was intragastric administration twice a day. After each group was instilled 12 h, 24 h, 6 rats were put to death to collect the required samples. We detected the expression of HMGB1 mRNA with fluorescent quantitation PCR (FQ-PCR), detected the expression of HMGB1 with Western blot. Mean comparison of each group's index all adopted SPSS 19.0 software for single factor variance analysis, P<0.05, the difference was statistically significant. Results At 12 h after instilled, the grayscale value of HMGB1 mRNA of model group (1.365 5±0.449 2) were higher than the control group (0.389 8±0.169 6) (P<0.01); the prednisone group (0.600 8±0.212 8), the Lianggesan decoction high group (0.670 7±0.217 5), middle group (0.731 5±0.200 1) and low dose group (0.799 3±0.552 6) were lower than the model group (P<0.01); the optical density value of HMGB1 of model group (30.03±0.15) were higher than the control group (12.20±0.10) (P<0.01); the prednisone group (13.47±0.25), the Lianggesan decoction high group (15.70±0.20), middle group (18.60±0.20) and low dose group (22.47±0.25) were lower than the model group (P<0.01); At 24 h after instilled, the relative grayscale value of HMGB1 mRNA of model group (1.710 9±0.802 8) were higher than the control group (0.691 8±0.245 3) (P<0.01); the prednisone group (0.727 0±0.097 7), the Lianggesan decoction high group (0.782 5±0.178 0), middle group (0.943 3±0.113 6) and low dose group (1.125 5±0.213 7) were lower than the model group (1.710 9±0.802 8) (P<0.01 or P<0.05); the relative optical density value of HMGB1 of model group (30.63±0.25) were higher than the control group (10.27±0.31) (P<0.01); the prednisone group (13.63±0.15), the Lianggesan decoction high group (18.63±0.21), middle group (22.57±0.21) and low dose group (24.53±0.32) were lower than the model group (30.03±0.15) (P<0.01 or P<0.05). Conclusion Lianggesan can repress the expression of HMGB1 and HMGB1 mRNA, lighten the inflammatory reaction of lung tissue in ALI.

Key words:Acute lung injury; Lianggesan; High mobility group proteins 1; High mobility group proteins 1 mRNA

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文献标引:周勇1 赵玮2.凉膈散对内毒素致急性肺损伤大鼠高迁移率族蛋白1影响[J/CD].中华临床医师杂志:电子版,2017,11(4):607.

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