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芪蛭皱肺颗粒对慢性阻塞性肺疾病模型大鼠TGF-β1、PDGF表达的调控作用
作者:张毅 李娟 张弢 苏韫 张晶 孙少伯 颜春鲁 赖乾 李金田 
单位:730000 兰州 甘肃中医药大学中医临床学院中医临床基础教研室(张毅、李娟、张弢、苏韫、张晶、孙少伯、颜春鲁、赖乾、李金田) 730000 兰州 敦煌医学与转化教育部重点实验室(李金田) 
关键词:慢性阻塞性肺疾病 转化生长因子β1 血小板衍生生长因子 芪蛭皱肺颗粒 
分类号:R256.14
出版年,卷(期):页码:2019,44(1):7-12
摘要:

 [摘要]  目的  观察芪蛭皱肺颗粒对慢性阻塞性肺疾病(COPD)模型大鼠肺组织转化生长因子β1(TGF-β1)及血小板衍生生长因子(PDGF)表达的影响,探讨芪蛭皱肺颗粒干预气道炎性反应及气道重塑的作用机制。方法  SPF Wistar大鼠60只,随机分为空白组(N)、模型组(M)、阳性对照组(D)和芪蛭皱肺颗粒高(HT)、中(MT )、低(LT)剂量组,每组10只。除空白组外,其余各组采用气管内滴注脂多糖(LPS)联合烟熏法复制大鼠COPD 模型。第29天开始模型组、空白组给予生理盐水1ml/100g,阳性对照组给予醋酸地塞米松溶液2.14μg/200g,芪蛭皱肺颗粒高、中、低剂量组分别给予芪蛭皱肺混悬液0.4320.2160.108g/200g,第58天时采用HE染色观察肺组织形态学的变化,反转录定量PCR(RT-qPCR)检测大鼠肺组织TGF-β1PDGF的转录水平,ELISAWestern blotting 法检测大鼠肺组织TGF-β1PDGF蛋白的表达水平。结果  各芪蛭皱肺颗粒干预组均可改善COPD大鼠的肺组织病理损害。RT-qPCR显示,与空白组比较,模型组大鼠肺组织TGF-β1PDGF mRNA表达量(分别为2.54±0.482.07±0.39)均明显升高(P0.05);与模型组比较,芪蛭皱肺颗粒高、中、低剂量组TGF-β1 mRNA表达量(分别为1.09±0.221.13±0.191.31±0.29)PDGF mRNA表达量(分别为1.22±0.161.38±0.251.44±0.18)均降低 (P0.05)ELISA结果显示,与空白组比较,模型组大鼠肺组织TGF-β1PDGF含量[分别为(315.80±19.20)pg/ml(105.37±10.84)pg/ml]升高(P0.05);与模型组比较,芪蛭皱肺颗粒高、低剂量组TGF-β1含量[分别为(228.60±11.20) pg/ml(253.01±18.02)pg/ml]降低(P0.05),芪蛭皱肺颗粒高、中剂量组PDGF含量[分别为(77.25±8.90)pg/ml(82.36±9.54)pg/ml]降低(P0.05)Western blotting结果显示,与空白组比较,模型组TGF-β1PDGF蛋白表达水平[分别为(1.55±0.15)pg/ml(1.48±0.16)pg/ml]升高(P0.05);与模型组比较,芪蛭皱肺颗粒高、中剂量组TGF-β1 PDGF蛋白表达水平[分别为(1.21±0.08)pg/ml(1.35±0.15)pg/ml(1.15±0.11)pg/ml(1.18±0.13)pg/ml]降低 (P0.05)结论  芪蛭皱肺颗粒可能通过降低COPD大鼠肺组织TGF-β1PDGF的表达,抑制肺组织炎性反应,从而干预气道重塑的发生,这可能是其防治COPD的机制之一。

[Abstract]  Objective  To observe the effect of Qizhi Zhoufei granules on the expression of transforming growth factor-β1 (TGF-β1) and platelet-derived growth factor (PDGF) in lung tissue of rats with chronic obstructive pulmonary disease (COPD), and to explore the mechanism of the effect of Qizhi Zhoufei granules on airway inflammation and airway remodeling. Methods  The COPD model was established in 60 SPF Wistar rats by intratracheal instillation of lipopolysaccharide (LPS) and smoking. Drug intervention began on the 29th day. The rats were randomly divided into blank group (N), model group (M), positive control group (D) and Qizhi Zhoufei granules high (HT), medium (MT) and low (LT) dose groups (10 rats in each group). On the 58th day, the changes of lung morphology were observed by hematoxylin eosin (HE) staining. The transcription level of TGF-β1, PDGF in rat lung tissue was detected by Q-PCR and the protein expression of TGF-β1, PDGF in rat lung tissue was detected by ELISA and Western blotting. Results  Qizhi Zhoufei granules intervention group can improve the lung tissue pathological damage of COPD rats. Q-PCR showed that compared with control group, the expression of TGF-β1 mRNA (2.54±0.48), PDGF mRNA (2.07±0.39) in model group significantly higher than that in control group (P<0.05); compared with model group, the mRNA levels of TGF-β1 (1.09±0.22, 1.13±0.19, 1.31±0.29) and PDGF (1.22±0.16, 1.38±0.25, 1.44±0.18) in the HT, MT and LT all decreased (P<0.05). ELISA showed that compared with control group, the levels of TGF-β1 [(315.80±19.20)pg/ml] and PDGF [(105.37±10.84)pg/ml] in the lungs of model group increased (P<0.05); compared with model group, the levels of TGF-β1 [(228.60±11.20)pg/ml], [(253.01±18.02)pg/ml] in the HT and LT all decreased (P<0.05), and the PDGF [(77.25±8.90) pg/ml, (82.36±9.54)pg/ml] levels in the HT and MT decreased (P<0.05). Western blotting showed that compared with control group, the protein expression of TGF-β1 [(1.55±0.15)pg/ml] and PDGF [(1.48±0.16)pg/ml] in model group significantly higher than that in control group (P<0.05); compared with model group, the protein expression levels of TGF-β1 [(1.21±0.08) pg/ml, (1.35±0.15)pg/ml] and PDGF [(1.15±0.11)pg/ml, (1.18±0.13)pg/ml] in the HT and MT decreased (P<0.05). Conclusion  The Qizhi Zhoufei granules may reduce the expression of TGF-β1 and PDGF in lung tissue of COPD rats, inhibit the inflammatory response of lung tissue and thus interfere with the occurrence of airway remodeling. This may be one of its mechanisms for preventing and treating COPD.

基金项目:
国家自然科学基金(81460709);甘肃省科技支甘计划(1011JKCA175)
作者简介:
730000 兰州 甘肃中医药大学中医临床学院中医临床基础教研室(张毅、李娟、张弢、苏韫、张晶、孙少伯、颜春鲁、赖乾、李金田);730000 兰州 敦煌医学与转化教育部重点实验室(李金田)
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