文章摘要
余仙娟,沈亚青,徐俭朴,等.基于JAK-STAT信号通路研究保肺定喘汤干预COPD肺血管重构的分子机制[J].浙江中医药大学学报,2020,44(12):1210-1214, 1221.
基于JAK-STAT信号通路研究保肺定喘汤干预COPD肺血管重构的分子机制
Study on the Molecular Mechanism of Baofei Dingchuan Decoction in Intervention of COPD Pulmonary Vascular Remodeling Based on JAK-STAT Signaling Pathway
投稿时间:2020-05-13  
DOI:10.16466/j.issn1005-5509.2020.12.014
中文关键词: COPD  肺血管重构  JAK/STAT  保肺定喘汤  CSE  PASMC  PCNA
英文关键词: COPD  pulmonary vascular remodeling  JAK/STAT  Baofei Dingchuan Decoction  CSE  PASMC  PCNA
基金项目:国家自然科学基金项目(81403336)
作者单位E-mail
余仙娟 浙江省中西医结合医院 杭州 310003  
沈亚青 浙江省中西医结合医院 杭州 310003  
徐俭朴 浙江省中西医结合医院 杭州 310003  
何飞 浙江省中西医结合医院 杭州 310003 lung1024@163.com 
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中文摘要:
      [目的]研究保肺定喘汤对慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)肺血管重构的两面神激酶/信号转导与转录激活子(Janus kinase/signal transducer and activator of transcriptions,JAK/STAT)信号通路的影响及相关分子机制。[方法]选取清洁级SD大鼠8只,随机分成2组,分别给予12.4g/(kg·d)保肺定喘汤及等量0.9%氯化钠溶液灌胃,末次灌胃后大鼠心脏采血并获取空白血清、含药血清。取大鼠肺动脉平滑肌细胞(pulmonary arterial smooth muscle cell,PASMC),空白对照组、空白血清组及药物血清组分别采用含有0.9%氯化钠、空白血清、含药血清的培养基培养,以流式细胞仪检测细胞周期。采用香烟提取物(cigarette smoke extract,CSE)进行诱导后分为模型组(培养基中加入10%CSE+空白血清)、药物组(培养基中加入10%CSE+10%含药血清),另设有对照组(培养基中加入0.9%氯化钠溶液+空白血清),以流式细胞术检测以上各组细胞凋亡率。将细胞分为对照组(培养基中加入0.9%氯化钠溶液+空白血清)、AG490组(培养基中加入终浓度为30nmol·mL-1的AG490)、模型组(培养基中加入10%CSE+空白血清)、CSE+AG490组(培养基中加入10%CSE+终浓度为30nmol·mL-1的AG490)、药物组(培养基中加入10%CSE+10%含药血清),以Western blot检测以上各组细胞增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)的蛋白表达,以及对照组、模型组、药物组JAK、STAT蛋白及磷酸化JAK、STAT蛋白表达。[结果]空白对照组、空白血清组及药物血清组的PASMC细胞周期分布比较差异均无统计学意义(P>0.05)。模型组PASMC凋亡率显著高于对照组(P<0.05),药物组PASMC凋亡率显著低于模型组(P<0.05)。模型组PCNA相对表达量显著高于药物组、CSE+AG490组及对照组(P<0.05),药物组与CSE+AG490组PCNA相对表达量差异无统计学意义(P>0.05)。模型组p-JAK/JAK、p-STAT1/STAT1、p-STAT3/STAT3均显著高于对照组(P<0.05),药物组p-JAK/JAK、p-STAT1/STAT1、p-STAT3/STAT3水平显著低于模型组(P<0.05)。[结论]保肺定喘汤能够抑制COPD的肺血管重构,其作用机制可能与抑制JAK、STAT1、STAT3等磷酸化有关。
英文摘要:
      [Objective] To study the effect of Baofei Dingchuan decoction on the Janus kinase/signal transducer and activator of transcriptions(JAK/STAT) signaling pathway of pulmonary vascular remodeling in chronic obstructive pulmonary disease(COPD) and its related molecular mechanism. [Methods] Eight SD rats with clean grade were randomly divided into two groups and given 12.4g/(kg·d) Baofei Dingchuan decoction and 0.9% sodium chloride solution, respectively. After the last gavage, blood was collected from the heart, and then blank serum and drug-containing serum were obtained. Pulmonary arterial smooth muscle cell(PASMC) was extracted from rats, 0.9% sodium chloride, blank serum and drug-containing serum were used for culture, respectively, as blank control group, blank serum group and drug serum group. Cell cycle was measured by flow cytometry. After induced with cigarette smoke extract(CSE), PASMC was divided into model group(containing 10%CSE+blank serum), drug group(containing 10%CSE+10% drug containing serum), and control group(containing 0.9% sodium chloride+blank serum), and the apoptotic rate was detected by flow cytometry in above groups. PASMC was furtherly divided into control group(containing 0.9% sodium chloride+blank serum), AG490 group(containing 30nmol·mL-1 AG490), model group(containing 10%CSE+blank serum), CSE+AG490 group(containing 10%CSE+30nmol·mL-1 AG490), drug group(containing 10%CSE+10% drug containing serum), Western blot was used to detect proliferating cell nuclear antigen(PCNA) expression in the above groups, and Janus kinase(JAK), signal transducer and activator of transcriptions(STAT), phosphorylated JAK(p-JAK) and phosphorylated STAT(p-STAT) expression in control group, model group and drug group. [Results] There was no statistically significant difference in PASMC cell cycle distribution among blank control group, blank serum group and drug serum group(P>0.05). The apoptosis rate of PASMC in model group was significantly higher than that in control group(P<0.05), and that in drug group was significantly lower than that in model group(P<0.05). The relative expression of PCNA in model group was significantly higher than that in drug group, CSE+AG490 group and control group(P<0.05), and there was no significant difference between drug group and CSE+AG490 group(P>0.05). The p-JAK/JAK, p-STAT1/STAT1 and p-STAT3/STAT3 levels in model group were significantly higher than those in control group(P<0.05), and the p-JAK/JAK, p-STAT1/STAT1 and p-STAT3/STAT3 levels in drug group were significantly lower than those in model group(P<0.05). [Conclusion] Baofei Dingchuan decoction can inhibit pulmonary vascular remodeling of COPD, and its mechanism may be related to the inhibition of phosphorylation of JAK, STAT1 and STAT3.
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