清金化痰汤通过中性粒细胞弹性蛋白酶干预AECOPD气道粘液高分泌的临床及实验研究
发布时间：2019-06-21 02:30
【摘要】：实验研究目的探索清金化痰汤在慢阻肺模型大鼠中性粒细胞弹性蛋白酶(Neutrophil elastase,NE)-表皮生长因子受体(epidermal growth factor receptor, EGFR)-有丝分裂原激活蛋白激酶(Mitogen-activated protein kinase, MAPK)-粘蛋白MUC5AC分泌”信号通路中的作用靶点。方法气道滴注脂多糖联合烟熏的方法建立慢性阻塞性肺疾病模型。50只清洁级Wistar大鼠随机分为空白对照组、模型组、清金化痰汤组、克拉霉素组与解毒清肺组。模型组、清金化痰汤组、克拉霉素组、解毒清肺组分别给予0.9%氯化钠溶液、清金化痰汤、克拉霉素片、解毒清肺合剂灌胃,空白对照组正常喂养,连续30天。实验第31日取材,每组随机选取6只,HE染色观察肺组织病理形态及黏液腺体增生,阿尔新蓝-过碘酸雪夫染色观察杯状细胞增生,实时荧光定量PCR检测各组大鼠肺组织NE.EGFR. MUC5AC基因表达,免疫组化检测肺组织及气道上皮NE、P-EGFR、磷酸化细胞外信号调节激酶(phosphorylated extracellular signal-regulated kinase, P-ERK)、磷酸化c-Jun氨基末端激酶(phosphorylated c-Jun N-terminal kinase, P-JNK).磷酸化p38丝裂原活化蛋白激酶(P-p38MAPK)、MUC5AC蛋白表达。结果与空白对照组比较,模型组粘液腺体增生、杯状细胞数目、NEmRNA、 MUC5ACmRNA表达、气道上皮NE、P-EGFR、P-ERK、P-JNK、P-p38、MUC5AC蛋白表达均显著升高。与模型组比较,清金化痰汤组粘液腺体增生、杯状细胞数目、NEmRNA、EGFRmRNA、MUC5ACmRNA、气道上皮NE、P-ERK、P-p38、MUC5AC蛋白表达均显著降低,且抑制NEmRNA、EGFRmRNA、MUC5ACmRNA、NE蛋白表达作用优于克拉霉素组。与模型组比较,清金化痰汤组大鼠气道上皮P-JNK表达显著升高,而克拉霉素组P-JNK表达显著降低,差异具有统计学意义。与模型组比较,清金化痰汤及克拉霉素对慢阻肺模型大鼠气道上皮P-EGFR蛋白表达均无影响。结论 清金化痰汤可能通过抑制NE及EGFR下游信号通路中ERK、p38信号通路,干预慢阻肺气道粘液高分泌。临床研究目的探索清金化痰汤通过NE调节慢阻肺急性加重期(acute exacerbation of chronic obstructive pulmonary disease, AECOPD,简称慢阻肺急性加重)(痰热壅肺证)气道粘液高分泌的作用。方法将符合纳入标准的慢阻肺急性加重期(痰热壅肺证)患者应用随机数字表法随机分为治疗组和对照组,共纳入病例156例,脱落21例,资料完整、具有统计学意义病例135例,治疗组71例,对照组64例。对照组在慢阻肺急性加重期常规西医治疗方案基础上,予以痰热清注射液30ml静脉滴注,每日1次,治疗组在常规西医治疗方案基础上,予以清金化痰汤颗粒剂口服,每日2次,每次1袋,疗程7天。观察指标：(1)治疗前检测患者动脉血气PaO2、PaCO2、SpO2、PH值；(2)分别在治疗第1、7天观察咳嗽频次、痰粘稠度、痰色、痰量、喘息、胸闷、呼吸困难症状积分及血常规白细胞总数、中性粒细胞百分比、C反应蛋白(C-reactionprotein, CRP)等实验室指标；(3)Elisa法检测患者治疗第1、7天血清NE、MUC5AC含量。结果(1)135例慢阻肺急性加重发病人群中,男性67人(49.63%),女性68人(50.37%)；(2)135例慢阻肺急性加重发病人群中,最小年龄46岁,最大年龄97岁,平均年龄(75.81±9.88)岁,其中在81-90岁分布最多(43.7%),其余依次是71-80岁(28.15%)、61-70岁(17.78%)、51-60岁(7.41%)、90岁(2.22%)、40-50岁(0.74%)；(3)135例慢阻肺急性加重发病人群中,有吸烟史77人(57.04%),无吸烟史58人(42.97%)；(4)135例慢阻肺急性加重发病人群既往病史中,从高到低依次是高血压病(52.59%)、冠状动脉粥样硬化性心脏病(45.93%)、糖尿病(22.96%)、高脂血症(20.74%)、反流性食管炎(17.78%)、脂肪肝(16.30%)、心律失常(12.59%)、脑梗死(11.11%)、肾功能不全(8.15%)、胃炎(7.41%)、甲状腺疾病(4.44%)；(5)135例慢阻肺急性加重住院人群中,肺功能Ⅲ级人群最多(45.19%),其余依次是肺功能Ⅱ级人群(34.07%)、肺功能Ⅰ级人群(20.74%)；(6)与治疗前相比,两组治疗后咳嗽频次、痰色、痰质、痰量、喘息、胸闷、mMRC评分、口干、小便黄、大便干单项症状积分均显著降低(P0.01),且治疗组在改善慢阻肺患者咳嗽频次、痰色、痰质、痰量方面显著优于对照组(P0.01)；(7)与治疗前相比,两组治疗后血清NE、MUC5AC含量、N%、CRP均显著降低(P0.01),且治疗组在减少黏蛋白MUC5AC分泌方面,作用显著优于对照组(P0.01)。与治疗前相比,治疗组WBC治疗后显著降低(P0.01),而对照组治疗后WBC与治疗前相比无差异(P0.05)；(8)与治疗前相比,两组治疗后症状总积分均显著降低(P0.01),且治疗组作用优于对照组(P0.01)。结论清热化痰类中药(清金化痰汤、痰热清)联合西药可以显著改善慢阻肺急性加重患者临床症状及实验室指标。清金化痰汤联合西药可以改善慢阻肺急性加重患者临床症状,尤其在咳嗽频次、痰粘稠度、痰色、痰量方面疗效显著,优于痰热清联合西药治疗组。清金化痰汤联合西药可以显著降低慢阻肺急性加重患者血清NE、MUC5AC. WBC、N%、CRP水平,且降低MUC5AC作用显著优于痰热清联合西药治疗组。
[Abstract]:Objective To explore the role of Qingjin Huatan Decoction in the rat neutrophil elastase (NE)-epidermal growth factor receptor (EGFR)-mitogen-activated protein kinase (EGFR)-mitogen-activated protein kinase (EGFR)-mitogen-activated protein kinase (EGFR)-mitogen-activated protein kinase (EGFR) in slow-resistance lung model rats. Methods A model of chronic obstructive pulmonary disease was established by the method of combined smoking with lipopolysaccharides in the airway. Fifty-one clean-grade Wistar rats were randomly divided into the blank control group, the model group, the Qingjin Huatan Decoction group, The Clarithromycin group and the Jiedu Qingfei Group. The model group, the Qingjin Huatan Decoction group, the Clarithromycin group, the Jiedu Qingfei group were given 0.9% sodium chloride solution, the Qingjin Huatan Decoction, the Clarithromycin Tablets, the Jiedu Qingfei Mixture, and the blank control group was normally fed for 30 days. The experiment was made on the 31st of the experiment. The expression of NE, EGFR and MUC5AC in the lung tissues of each group was detected by real-time fluorescence quantitative PCR. The expression of NE, EGFR and MUC5AC in the lung of each group was detected by real-time fluorescence quantitative PCR. P-EGFR, phosphorylated extracellular signal-regulated kinase (P-ERK), phosphorylated c-Jun amino terminal kinase (P-JNK), phosphorylated p38 mitogen-activated protein kinase (P-p38 MAPK), MUC5AC protein expression. The number of goblet cells, the expression of NEmRNA, MUC5ACmRNA, airway epithelium NE, P-EGFR, P-ERK, P-JNK, P-p38 and MUC5AC were all significantly increased. Compared with the model group, the expression of mucus glands, goblet cells, NEmRNA, EGFRmRNA, MUC5ACmRNA, airway epithelium NE, P-ERK, P-p38, The expression of MUC5AC was significantly lower, and the expression of NEmRNA, EGFRmRNA, MUC5ACmRNA and NE protein was better than that of clarithromycin. Conclusion Qingjin Huatan Decoction can inhibit the signal pathway of ERK and p38 in the downstream signal pathway of NE and EGFR, compared with the model group. The purpose of the clinical study was to explore the acute exacerbation of chronic obstructive pulmonary disease (AECOPD) through NE. The method is to randomly divide into treatment group and control group according to the application of random digital table method in the patients with acute exacerbation of chronic obstructive pulmonary disease (phlegm-heat and lung syndrome), which are included in the standard. The data was complete,135 cases with statistical significance,71 in the treatment group and 64 in the control group. The control group, on the basis of the conventional western medicine treatment plan of the acute exacerbation of the chronic obstructive pulmonary disease, was given a 30-ml intravenous drip of the phlegm-heat-clearing injection, once a day, and the treatment group was on the basis of the conventional western medicine treatment plan. The preparation method comprises the following steps: (1) detecting the PaO2, PaCO2, SpO2 and PH value of the arterial blood gas of the patient before treatment; (2) observing the cough frequency, the phlegm viscosity, the phlegm color, the sputum volume and the wheezing after the treatment on the first and the seventh day respectively; Results (1) The serum NE and MUC5AC content in the first and 7th day of the treatment were detected by the Elisa method. Results (1)135 patients with acute exacerbation of chronic obstructive pulmonary disease, 67 (49.63%) males and 68 (50.37%) females; (2)135 patients with acute exacerbation of chronic obstructive pulmonary disease, with a minimum age of 46, a maximum age of 97, an average age (75.81 to 9.88), with a maximum distribution of (43.7%) at the age of 81 to 90; The rest were 71-80 years (28.15%),61-70 years (17.78%),51-60 years (7.41%),90-year-old (2.22%),40-50 years (0.74%), (3)135 patients with chronic obstructive pulmonary disease with a history of 77 (57.04%), no smoking history of 58 (42.97%), (4)135 patients with chronic obstructive pulmonary disease with prior medical history, From high to low, hypertension (52.59%), coronary atherosclerotic heart disease (45.93%), diabetes (22.96%), hyperlipidemia (20.74%), reflux esophagitis (17.78%), fatty liver (16.30%), arrhythmia (12.59%), and cerebral infarction (11.11%). Renal insufficiency (8.15%), gastritis (7.41%), thyroid disease (4.44%), (5)135 patients with chronic obstructive pulmonary acute exacerbation, the lung function class 鈪，
本文编号：2503715
[Abstract]:Objective To explore the role of Qingjin Huatan Decoction in the rat neutrophil elastase (NE)-epidermal growth factor receptor (EGFR)-mitogen-activated protein kinase (EGFR)-mitogen-activated protein kinase (EGFR)-mitogen-activated protein kinase (EGFR)-mitogen-activated protein kinase (EGFR) in slow-resistance lung model rats. Methods A model of chronic obstructive pulmonary disease was established by the method of combined smoking with lipopolysaccharides in the airway. Fifty-one clean-grade Wistar rats were randomly divided into the blank control group, the model group, the Qingjin Huatan Decoction group, The Clarithromycin group and the Jiedu Qingfei Group. The model group, the Qingjin Huatan Decoction group, the Clarithromycin group, the Jiedu Qingfei group were given 0.9% sodium chloride solution, the Qingjin Huatan Decoction, the Clarithromycin Tablets, the Jiedu Qingfei Mixture, and the blank control group was normally fed for 30 days. The experiment was made on the 31st of the experiment. The expression of NE, EGFR and MUC5AC in the lung tissues of each group was detected by real-time fluorescence quantitative PCR. The expression of NE, EGFR and MUC5AC in the lung of each group was detected by real-time fluorescence quantitative PCR. P-EGFR, phosphorylated extracellular signal-regulated kinase (P-ERK), phosphorylated c-Jun amino terminal kinase (P-JNK), phosphorylated p38 mitogen-activated protein kinase (P-p38 MAPK), MUC5AC protein expression. The number of goblet cells, the expression of NEmRNA, MUC5ACmRNA, airway epithelium NE, P-EGFR, P-ERK, P-JNK, P-p38 and MUC5AC were all significantly increased. Compared with the model group, the expression of mucus glands, goblet cells, NEmRNA, EGFRmRNA, MUC5ACmRNA, airway epithelium NE, P-ERK, P-p38, The expression of MUC5AC was significantly lower, and the expression of NEmRNA, EGFRmRNA, MUC5ACmRNA and NE protein was better than that of clarithromycin. Conclusion Qingjin Huatan Decoction can inhibit the signal pathway of ERK and p38 in the downstream signal pathway of NE and EGFR, compared with the model group. The purpose of the clinical study was to explore the acute exacerbation of chronic obstructive pulmonary disease (AECOPD) through NE. The method is to randomly divide into treatment group and control group according to the application of random digital table method in the patients with acute exacerbation of chronic obstructive pulmonary disease (phlegm-heat and lung syndrome), which are included in the standard. The data was complete,135 cases with statistical significance,71 in the treatment group and 64 in the control group. The control group, on the basis of the conventional western medicine treatment plan of the acute exacerbation of the chronic obstructive pulmonary disease, was given a 30-ml intravenous drip of the phlegm-heat-clearing injection, once a day, and the treatment group was on the basis of the conventional western medicine treatment plan. The preparation method comprises the following steps: (1) detecting the PaO2, PaCO2, SpO2 and PH value of the arterial blood gas of the patient before treatment; (2) observing the cough frequency, the phlegm viscosity, the phlegm color, the sputum volume and the wheezing after the treatment on the first and the seventh day respectively; Results (1) The serum NE and MUC5AC content in the first and 7th day of the treatment were detected by the Elisa method. Results (1)135 patients with acute exacerbation of chronic obstructive pulmonary disease, 67 (49.63%) males and 68 (50.37%) females; (2)135 patients with acute exacerbation of chronic obstructive pulmonary disease, with a minimum age of 46, a maximum age of 97, an average age (75.81 to 9.88), with a maximum distribution of (43.7%) at the age of 81 to 90; The rest were 71-80 years (28.15%),61-70 years (17.78%),51-60 years (7.41%),90-year-old (2.22%),40-50 years (0.74%), (3)135 patients with chronic obstructive pulmonary disease with a history of 77 (57.04%), no smoking history of 58 (42.97%), (4)135 patients with chronic obstructive pulmonary disease with prior medical history, From high to low, hypertension (52.59%), coronary atherosclerotic heart disease (45.93%), diabetes (22.96%), hyperlipidemia (20.74%), reflux esophagitis (17.78%), fatty liver (16.30%), arrhythmia (12.59%), and cerebral infarction (11.11%). Renal insufficiency (8.15%), gastritis (7.41%), thyroid disease (4.44%), (5)135 patients with chronic obstructive pulmonary acute exacerbation, the lung function class 鈪，
本文编号：2503715
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