调补肺肾方药对PM2.5暴露大鼠鼻腔黏液高分泌的保护作用
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摘要
目的:采用R值综合评价法多指标综合评价调补肺肾系列方药(补肺方、补肺健脾方、补肺益肾方、益气滋肾方)对PM2.5暴露大鼠鼻腔黏液高分泌的调节作用。方法:将60只健康雄性SD大鼠随机分为对照组、模型组、补肺方组、补肺健脾方组、补肺益肾方组及益气滋肾方组,每组10只。采用大气PM2.5实时浓缩暴露制备大鼠模型。并自暴露第1天起,各组大鼠分别给予0.9%氯化钠溶液、补肺方、补肺健脾方、补肺益肾方及益气滋肾方灌胃,持续14d,于第15天取得各组大鼠鼻腔组织,分别采用阿利辛蓝-过碘酸-雪夫(AB-PAS)法和免疫组化法测定各组大鼠鼻黏膜上皮杯状细胞数量及黏蛋白MUC5ac、MUC5b的表达情况。采用R值综合评价法,对调补肺肾系列方药调节PM2.5诱导的鼻腔黏液高分泌进行综合评价。结果:与对照组比较,模型组大鼠鼻黏膜上皮杯状细胞比例显著升高(P<0.01),且MUC5ac、MUC5b水平显著升高(P<0.01);与模型组比较,4个治疗组大鼠鼻黏膜上皮杯状细胞比例及MUC5ac、MUC5b水平显著降低(P<0.01),综合纠正强度排序为:补肺健脾方、补肺益肾方、补肺方、益气滋肾方。结论:PM2.5暴露可诱导鼻黏膜上皮黏液分泌增加,调补肺肾系列方药对其具有明显调节作用,其中以补肺健脾方、补肺益肾方效果更佳。
Objective: To evaluate the effects of series formulae for invigorating lung and kidney(Bufei Formula, Bufei Jianpi Formula, Bufei Yishen Formula and Yiqi Zishen Formula) on nasal mucus hypersecretion in rats exposed to PM2.5 by R-value comprehensive evaluation. Methods: Sixty male SD rats were randomly divided into six groups(10 rats in each group):control group, model group, Bufei Formula group, Bufei Jianpi Formula group, Bufei Yishen Formula group and Yiqi Zishen Formula group. Real-time atmospheric PM2.5 after concentrated were used to establish rats model. Since the first day exposed to PM2.5, rats were gavaged with saline in control groups, Bufei Formula, Bufei Jianpi Formula, Bufei Yishen Formula, and Yiqi Zishen Formula respectively in intervention groups for two weeks. The nasal mucosa tissues were collected at 15 th day.Alcian Blue/PAS(AB-PAS) staining method was used to detect the quantity of goblet cells in the nasal mucous tissues. The immunohistochemistry method was used to measure the expressions of MUC5 ac and MUC5 b. R-value comprehensive evaluation was adopted to evaluate the effect on nasal mucus hypersecretion. Results: Compared with the control group, the rate of goblet cells and the expressions of MUC5 ac, MUC56 in the model group were significantly increased, as well as secretion of mucus.The four treatment groups can significantly alleviate goblet cell rate and decrease the expressions of MUC5 ac and MUC5 b in the nasal mucous tissues compared with the model group, and the order of corrective effect intensity of which was Bufei Jianpi Formula, Bufei Yishen Formula, Bufei Formula and Yiqi Zishen Formula. Conclusion: PM2.5 exposure can lead to nasal mucus hypersecretion, while the series formulae for invigorating lung and kidney have significant regulating effect on it. Bufei Jianpi Formula and Bufei Yishen Formula are the most effective formulas.
Objective: To evaluate the effects of series formulae for invigorating lung and kidney(Bufei Formula, Bufei Jianpi Formula, Bufei Yishen Formula and Yiqi Zishen Formula) on nasal mucus hypersecretion in rats exposed to PM2.5 by R-value comprehensive evaluation. Methods: Sixty male SD rats were randomly divided into six groups(10 rats in each group):control group, model group, Bufei Formula group, Bufei Jianpi Formula group, Bufei Yishen Formula group and Yiqi Zishen Formula group. Real-time atmospheric PM2.5 after concentrated were used to establish rats model. Since the first day exposed to PM2.5, rats were gavaged with saline in control groups, Bufei Formula, Bufei Jianpi Formula, Bufei Yishen Formula, and Yiqi Zishen Formula respectively in intervention groups for two weeks. The nasal mucosa tissues were collected at 15 th day.Alcian Blue/PAS(AB-PAS) staining method was used to detect the quantity of goblet cells in the nasal mucous tissues. The immunohistochemistry method was used to measure the expressions of MUC5 ac and MUC5 b. R-value comprehensive evaluation was adopted to evaluate the effect on nasal mucus hypersecretion. Results: Compared with the control group, the rate of goblet cells and the expressions of MUC5 ac, MUC56 in the model group were significantly increased, as well as secretion of mucus.The four treatment groups can significantly alleviate goblet cell rate and decrease the expressions of MUC5 ac and MUC5 b in the nasal mucous tissues compared with the model group, and the order of corrective effect intensity of which was Bufei Jianpi Formula, Bufei Yishen Formula, Bufei Formula and Yiqi Zishen Formula. Conclusion: PM2.5 exposure can lead to nasal mucus hypersecretion, while the series formulae for invigorating lung and kidney have significant regulating effect on it. Bufei Jianpi Formula and Bufei Yishen Formula are the most effective formulas.
引文
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[10]Li J S,Li Y,Li SY,et al.Long-term effects of Tiaobu Feishen therapies on systemic and local inflammation responses in rats with stable chronic obstructive pulmonary disease.Journal of Integrative Medicine,2012,10(9):1039-1048
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[12]田燕歌,崔红新,李建生,等.调补肺肾三法调控慢性阻塞性肺疾病稳定期大鼠炎症反应的R值综合评价.中医杂志,2015,56(15):1330-1334
[13]黄继汉,黄晓辉,陈志扬,等.药理试验中动物间和动物与人体间的等效剂量换算.中国临床药理学与治疗学,2004,9(9):1069-1072
[14]Jack R Harkema,Stephan A Carey,James G Wagner.The nose revisited:A brief review of the comparative structure,function,and toxicologic pathology of the nasal epithelium.toxicologic pathology.Toxicol Pathol,2006,34(3):252-269
[15]Mery S,Gross E A,Joyner D R,et al.Nasal diagrams:A tool for recording the distribution of nasal lesions in rats and mice.Toxicol Pathol,1994,22(4):353-372
[16]Livraghi-Butrico A,Grubb B R,Wilkinson K J,et al.Contribution of mucus concentration and secreted mucins Muc5ac and Muc5b to the pathogenesis of muco-obstructive lung disease.Mucosal Immunol,2017,10(3):829
[17]Bonser L R,Zlock L,Finkbeiner W,et al.Epithelial tethering of MUC5AC-rich mucus impairs mucociliary transport in asthma.J Clin Invest,2016,126(6):2367-2371
[18]Torricelli A A,Matsuda M,Novaes P,et al.Effects of ambient levels of traffic-derived air pollution on the ocular surface:Analysis of symptoms,conjunctival goblet cell count and mucin 5AC gene expression.Environ Res,2014,131:59-63
[19]Kim S S,Kim C H,Kim J W,et al.Airborne particulate matter increases MUC5AC expression by downregulating Claudin-1expression in human airway cells.BMB Rep,2017,50(10):516-521
[20]Okuda K,Chen G,Subramani D B,et al.Localization of secretory mucins MUC5AC and MUC5B in normal/healthy human airways.Am J Respir Crit Care Med,2018:1-92
[21]Liu Z,Geng W,Jiang C,et al.Hydrogen-rich saline inhibits tobacco smoke-induced chronic obstructive pulmonary disease by alleviating airway inflammation and mucus hypersecretion in rats.Exp Biol Med(Maywood),2017,242(15):1534-1541
[22]Mogensen C,Tos M.Density of goblet cells in the normal adult human nasal septum.Anat Anz,1977,141(3):237-247
[23]中华中医药学会内科分会肺系病专业委员会.慢性阻塞性肺疾病中医证候诊断标准(2011版).中医杂志,2012,53(2):177-178
[24]中华中医药学会内科分会肺系病专业委员会.慢性阻塞性肺疾病中医诊疗指南(2011版).中医杂志,2012,53(1):80-84
[25]李亚,王元元,李建生,等.调补肺肾三法对COPD大鼠肺组织基质金属蛋白酶的影响及远后效应.北京中医药大学学报,2012,35(9):615-619
[2]Maji K J,Dikshit A K,Arora M,et al.Estimating premature mortality attributable to PM2.5 exposure and benefit of air pollution control policies in China for 2020.Sci Total Environ,2018,612:683-693
[3]Fan J,Li S,Fan C,et al.The impact of PM2.5 on asthma emergency department visits:A systematic review and meta-analysis.Environ Sci Pollut Res Int,2016,23(1):843-850
[4]Vawda S,Mansour R,Takeda A,et al.Associations between inflammatory and immune response genes and adverse respiratory outcomes following exposure to outdoor air pollution:A HuGE systematic review.Am J Epidemiol,2014,179(4):432-442
[5]李建生.正虚积损为慢性阻塞性肺疾病的主要病机.中华中医药杂志,2011,26(8):710-713
[6]张海龙.辨证治疗慢性阻塞性肺疾病稳定期的疗效及对全身炎症反应的影响.郑州:河南中医学院,2010
[7]李建生,余学庆,王明航,等.中医治疗慢性阻塞性肺疾病研究的策略与实践.中华中医药杂志,2012,27(6):1607-1614
[8]Li S Y,Li J S,Wang M H,et al.Effects of comprehensive therapy based on traditional Chinese medicine patterns in stable chronic obstructive pulmonary disease:A four-center,open-label,randomized,controlled study.BMC Complement Altern Med,2012,12:197
[9]Li J S,Li S Y,Xie Y,et al.The effective evaluation on symptoms and quality of life of chronic obstructive pulmonary disease patients treated by comprehensive therapy based on traditional Chinese medicine patterns.Complement Ther Med,2013,21(6):595-602
[10]Li J S,Li Y,Li SY,et al.Long-term effects of Tiaobu Feishen therapies on systemic and local inflammation responses in rats with stable chronic obstructive pulmonary disease.Journal of Integrative Medicine,2012,10(9):1039-1048
[11]田燕歌,李亚,李建生,等.调补肺肾三法对慢性阻塞性肺疾病大鼠肺组织氧化应激的影响和远后效应.中华中医药杂志,2014,29(2):621-624
[12]田燕歌,崔红新,李建生,等.调补肺肾三法调控慢性阻塞性肺疾病稳定期大鼠炎症反应的R值综合评价.中医杂志,2015,56(15):1330-1334
[13]黄继汉,黄晓辉,陈志扬,等.药理试验中动物间和动物与人体间的等效剂量换算.中国临床药理学与治疗学,2004,9(9):1069-1072
[14]Jack R Harkema,Stephan A Carey,James G Wagner.The nose revisited:A brief review of the comparative structure,function,and toxicologic pathology of the nasal epithelium.toxicologic pathology.Toxicol Pathol,2006,34(3):252-269
[15]Mery S,Gross E A,Joyner D R,et al.Nasal diagrams:A tool for recording the distribution of nasal lesions in rats and mice.Toxicol Pathol,1994,22(4):353-372
[16]Livraghi-Butrico A,Grubb B R,Wilkinson K J,et al.Contribution of mucus concentration and secreted mucins Muc5ac and Muc5b to the pathogenesis of muco-obstructive lung disease.Mucosal Immunol,2017,10(3):829
[17]Bonser L R,Zlock L,Finkbeiner W,et al.Epithelial tethering of MUC5AC-rich mucus impairs mucociliary transport in asthma.J Clin Invest,2016,126(6):2367-2371
[18]Torricelli A A,Matsuda M,Novaes P,et al.Effects of ambient levels of traffic-derived air pollution on the ocular surface:Analysis of symptoms,conjunctival goblet cell count and mucin 5AC gene expression.Environ Res,2014,131:59-63
[19]Kim S S,Kim C H,Kim J W,et al.Airborne particulate matter increases MUC5AC expression by downregulating Claudin-1expression in human airway cells.BMB Rep,2017,50(10):516-521
[20]Okuda K,Chen G,Subramani D B,et al.Localization of secretory mucins MUC5AC and MUC5B in normal/healthy human airways.Am J Respir Crit Care Med,2018:1-92
[21]Liu Z,Geng W,Jiang C,et al.Hydrogen-rich saline inhibits tobacco smoke-induced chronic obstructive pulmonary disease by alleviating airway inflammation and mucus hypersecretion in rats.Exp Biol Med(Maywood),2017,242(15):1534-1541
[22]Mogensen C,Tos M.Density of goblet cells in the normal adult human nasal septum.Anat Anz,1977,141(3):237-247
[23]中华中医药学会内科分会肺系病专业委员会.慢性阻塞性肺疾病中医证候诊断标准(2011版).中医杂志,2012,53(2):177-178
[24]中华中医药学会内科分会肺系病专业委员会.慢性阻塞性肺疾病中医诊疗指南(2011版).中医杂志,2012,53(1):80-84
[25]李亚,王元元,李建生,等.调补肺肾三法对COPD大鼠肺组织基质金属蛋白酶的影响及远后效应.北京中医药大学学报,2012,35(9):615-619