XIANG Shi-jian, SHENG Hua-fang, LYU Ya-mei, et al. Effect of Dongbing Xiazhi Fang on Airway Inflammation and Intestinal Flora in Guinea Pig Model of Asthma Through Acupoint Application[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(9): 95-102.
DOI:
XIANG Shi-jian, SHENG Hua-fang, LYU Ya-mei, et al. Effect of Dongbing Xiazhi Fang on Airway Inflammation and Intestinal Flora in Guinea Pig Model of Asthma Through Acupoint Application[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(9): 95-102. DOI: 10.13422/j.cnki.syfjx.20180833.
Effect of Dongbing Xiazhi Fang on Airway Inflammation and Intestinal Flora in Guinea Pig Model of Asthma Through Acupoint Application
目的:探究冬病夏治方穴位贴敷给药对哮喘豚鼠的治疗作用及其对肠道菌群的影响。方法:将75只豚鼠随机分成空白组、哮喘模型组、阳性(地塞米松磷酸钠)组、穴位给药组、非穴位给药组(n=15),采用卵蛋白注射及雾化吸入的方法建立豚鼠哮喘模型,通过肺组织病理苏木素-伊红(HE),马松(Masson)染色观察冬病夏治方对豚鼠哮喘的治疗效果。提取豚鼠粪便总基因组DNA,对16 S rRNA V4可变区基因进行PCR扩增及Illumina HiSeq测序,将基因序列相似度高于97%的序列进行聚类归并,并进行生物信息学分析。结果:与哮喘模型组比较,各给药组豚鼠肺组织炎症细胞减少,支气管收缩、气道上皮增生症状改善明显,恢复良好,胶原纤维沉积均有不同程度减少,且地塞米松磷酸钠组与穴位给药组的减少程度明显,穴位给药组的治疗效果优于非穴位给药组。通过操作分类单元(OTU)及其丰度分析发现,冬病夏治方穴位贴敷给药后豚鼠的肠道菌群多样性升高,普氏菌属显著增多,拟杆菌门减少,变形菌增多;哮喘模型组厚壁菌数目较空白组及各给药组减少。结论:冬病夏治方穴位贴敷能有效治疗哮喘,改善哮喘症状,调控哮喘豚鼠肠道菌群的构成。提示该方可能通过调控肠道菌群从而影响机体免疫,达到治疗哮喘的效果,为研究哮喘的治疗机制提供一定的理论基础。
Abstract
Objective: To investigate the efficacy of Dongbing Xiazhi Fang (DXF) on asthmatic guinea pigs through acupoint-application
and its effect on intestinal flora of asthmatic guinea pigs. Method: Seventy five guinea pigs were randomly divided into blank control group
asthmatic model group
positive control group
acupoint-application group and non-acupoint-application group (n=15). The asthmatic model of guinea pigs was established through ovalbumin injection and aerosol inhalation
except for the blank control group. The hematoxylin-eosin(HE) and Masson staining pathologic sections of lung tissue in asthma were observed. The total genomic DNA was extracted from guinea pig feces
and the 16 S rRNA V4 variable region gene was amplified by PCR and sequenced by Illumina HiSeq. The gene sequences with the similarity of more than 97% were clustered
and analyzed by bioinformatics. Result: Compared with the asthmatic model group
the other groups of guinea pigs showed reduced inflammatory cells
bronchoconstriction
and alleviated airway epithelial hyperplasia and good recovery and various degrees of reduced collagen fiber deposition. The positive group and the acupoint-application group showed a more obvious degree of reduction
and the therapeutic effect of the acupoint-application group was better than that of the non-acupoint-application group. The analysis of operational taxonomic unit (OTU) and its abundance indicated that the proportion of various bacteria changed after the onset of asthma. The diversity of intestinal flora of guinea pigs significantly increased after being treated with DXF and Prevotella. The proportion of Bacteroides in the positive control group and other administration groups were less than that in the asthmatic model group
while the proportion of Proteobacteria and Firmicutes were more than that in the asthmatic model group. Conclusion: DXF has an effect in treating asthma through transdermal acupoint-application
and can alleviate asthma symptoms
regulate the composition of intestinal flora in asthmatic guinea pigs. The results suggest that the prescription have the efficacy on asthma by affecting the body immunity and regulating the intestinal flora
and provide a theoretical basis for studying the treatment mechanism of asthma.
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