五虎汤通过调控miRNA-146a改善幼龄哮喘大鼠气道重塑  

Improvement of Wuhu Decoction on airway remodeling in young asthmatic rats by regulating miRNA-146a

作　　者：邓羿駃 王孟清[1] 谢静[1] 胡燕[1] 曾洁 罗菁[2,3] 姚冰 DENG Yijue;WANG Mengqing;XIE Jing;HU Yan;ZENG Jie;LUO Jing;YAO Bing(The First Hospital of Hunan University of Chinese Medicine,Changsha,Hunan 410007,China;Hunan University of Chinese Medicine,Changsha,Hunan 410208,China;Shenzhen Nanshan District Shekou People's Hospital,Shenzhen,Guangdong 154100,China)

机构地区：[1]湖南中医药大学第一附属医院,湖南长沙410007 [2]湖南中医药大学,湖南长沙410208 [3]深圳市南山区蛇口人民医院,深圳154100

出　　处：《湖南中医药大学学报》2023年第5期799-806,共8页Journal of Hunan University of Chinese Medicine

基　　金：国家自然科学基金面上项目(82174437);湖南省自然科学基金面上项目(2022JJ30037);湖南中医药大学校级科研项目(2022XYLH014);湖南中医药大学研究生创新课题项目(2021CX28)。

摘　　要：目的探讨五虎汤通过调控miRNA-146a对幼龄哮喘大鼠气道重塑的影响。方法SPF级幼龄雌性SD大鼠80只,先随机将20只作为空白组,60只作为造模组。造模组采用卵清白蛋白联合氢氧化铝五点注射法致敏以及卵清白蛋白雾化激发哮喘。两组各随机抽取10只检测气道反应性,评判造模是否成功。造模成功后,剩下的10只空白组大鼠作为正常组,50只造模组大鼠随机分为5组(模型组、五虎汤低剂量组、五虎汤中剂量组、五虎汤高剂量组、地塞米松组),每组10只。五虎汤高、中、低剂量组分别灌胃五虎汤4.428、2.214、1.107 g/kg,地塞米松组灌胃地塞米松0.075 mg/kg,正常组及模型组灌胃等容积生理盐水,1次/d,连续2周后处理大鼠。气道反应性检测大鼠气道阻力;HE、PAS及Masson染色法分别观察大鼠肺组织气道炎症细胞浸润、气道黏液储备和气道胶原沉积情况;Western blot法检测基质金属蛋白酶-9(matrix metalloprotein-9,MMP-9)、金属蛋白酶组织抑制因子-1(tissue inhibitor of matrix metalloproteinases-1,TIMP-1)、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、转化生长因子-β1(transforming growth factor-β1,TGF-β1)蛋白表达水平;qPCR法检测miRNA-146a及MMP-9、TIMP-1 mRNA含量,并对miRNA-146a与MMP-9、TIMP-1 mRNA进行相关性分析。结果与空白组比较,造模组气道阻力显著升高(P<0.01),提示造模成功。与正常组比较,模型组大鼠肺组织周围炎性细胞浸润,上皮细胞化生,炎性黏液较多,气道壁增厚,气道胶原广泛沉积;肺组织α-SMA、TGF-β1、MMP-9、TIMP-1、miRNA-146a、MMP-9 mRNA、TIMP-1 mRNA均显著升高(P<0.01)。与模型组相比,五虎汤高、中、低剂量组及地塞米松组气道阻力明显降低(P<0.01),肺组织病理情况明显缓解,且五虎汤高剂量组及地塞米松组改善更加明显;α-SMA、TGF-β1、MMP-9、TIMP-1、miRNA-146a、MMP-9 mRNA、TIMP-1 mRNA均显著下降(P<0.01)。五虎汤中、低剂量�Objective To explore the effects of Wuhu Decoction(WHD)on airway remodeling in young asthmatic rats by regulating miRNA-146a.Methods A total of 80 young female SD rats of SPF grade were randomly divided into blank group(n=20)and modeling group(n=60).To stimulate asthma,the modeling group was sensitized by ovalbumin(OVA)combined with aluminum hydroxide five-point injection and OVA atomization.Ten rats were randomly selected from each group to detect airway responsiveness and evaluate the success of modeling.After successful modeling,the remaining 10 rats in the blank group were considered as the normal group,and 50 rats in the modeling group were randomly subdivided into 5 groups(model group,low-,medium-and high-dose WHD groups,and dexamethasone group),with 10 rats in each group.The high-,medium-and low-dose WHD groups were respectively given WHD 4.428,2.214,1.107 g/kg by gavage,the dexamethasone group was given dexamethasone 0.075 mg/kg by gavage,and the normal group and model group were given equal volume physiological saline by gavage,once a day,for 2 consecutive weeks.Then the rats were anaesthetized and tracheotomized.The airway resistance in rats was measured by airway responsiveness;the airway inflammatory cell infiltration,airway mucus reserve,and airway collagen deposition in lung tissue of rats were observed respectively by HE,PAS and Masson staining methods;the expression levels of the matrix metalloproteinase-9(MMP-9),tissue inhibitor of matrix metalloproteinases-1(TIMP-1),α-smooth muscle actin(α-SMA)and transforming growth factor-β1(TGF-β1)protein were determined by Western blot;the content of miRNA-146a,MMP-9,TIMP-1 mRNA was examined by qPCR and the correlation between miRNA-146a and MMP-9,TIMP-1 mRNA was analyzed.Results Compared with the blank group,the airway resistance in the modeling group was significantly higher(P<0.01),which indicated the success of modeling.Compared with the normal group,the rats in the model group demonstrated as follows:inflammatory cell infiltration around the lung ti

关 键 词：五虎汤 幼龄哮喘大鼠 miRNA-146a 气道重塑 基质金属蛋白酶-9 金属蛋白酶组织抑制因子-1 Α-平滑肌肌动蛋白 转化生长因子-β1 

分 类 号：R285.5[医药卫生—中药学]