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邓羿駃,王孟清,谢静,胡燕,曾洁,罗菁,姚冰.五虎汤通过调控miRNA-146a改善幼龄哮喘大鼠气道重塑[J].湖南中医药大学学报,2023,43(5):799-806[点击复制] |
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| 五虎汤通过调控miRNA-146a改善幼龄哮喘大鼠气道重塑 |
| 邓羿駃,王孟清,谢静,胡燕,曾洁,罗菁,姚冰 |
| (湖南中医药大学第一附属医院, 湖南 长沙 410007;湖南中医药大学, 湖南 长沙 410208;湖南中医药大学, 湖南 长沙 410208;深圳市南山区蛇口人民医院, 深圳 154100) |
| 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、低剂量组α-SMA、TGF-β1、miRNA-146a、MMP-9 mRNA、TIMP-1 mRNA及五虎汤低剂量组TIMP-1显著高于地塞米松组(P<0.05、P<0.01);五虎汤高、中剂量组α-SMA、TIMP-1、miRNA-146a、MMP-9 mRNA、TIMP-1 mRNA及五虎汤高剂量组TGF-β1低于五虎汤低剂量组(P<0.05、P<0.01);五虎汤高剂量组α-SMA、TGF-β1、miRNA-146a、TIMP-1 mRNA低于五虎汤中剂量组(P<0.05、P<0.01)。Pearson相关性分析显示,大鼠肺组织miRNA-146a与MMP-9 mRNA、TIMP-1 mRNA均有极强正相关性(r分别为0.956、0.973,P<0.01)。结论 五虎汤能够有效改善哮喘大鼠气道重塑,这可能与五虎汤抑制miRNA-146a表达,降低MMP-9、TIMP-1及其mRNA,以及减少α-SMA、TGF-β1相关。 |
| 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 |
| DOI：10.3969/j.issn.1674-070X.2023.05.007 |
| 投稿时间：2023-02-20 |
| 基金项目: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）。 |
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| Improvement of Wuhu Decoction on airway remodeling in young asthmatic rats by regulating miRNA-146a |
| 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;Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Shenzhen Nanshan District Shekou People's Hospital, Shenzhen, Guangdong 154100, China) |
| Abstract: |
| 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 tissue, epithelial cell metaplasia, more inflammatory mucus, airway wall thickening and extensive deposition of airway collagen; the α-SMA, TGF-β1, MMP-9, TIMP-1, miRNA-146a, MMP-9 mRNA and TIMP-1 mRNA expressions in lung tissue of rats in the model group were significantly higher (P<0.01). Compared with the model group, the airway resistance in high-, medium- and low-dose WHD groups and the dexamethasone group was significantly lower (P<0.01), and the pathological condition of lung tissue was significantly alleviated, moreover, the improvement in the high-dose WHD group and the dexamethasone group was more significant; the α-SMA, TGF-β1, MMP-9, TIMP-1, miRNA-146a, MMP-9mRNA, TIMP-1 mRNA expressions in high-, medium- and low-dose WHD groups and the dexamethasone group were significantly lower (P<0.01). The α-SMA, TGF-β1, miRNA-146a, MMP-9 mRNA, and TIMP-1 mRNA expressions in medium- and low-dose WHD groups and the TIMP-1 of the rats in low-dose WHD group were significantly higher than those in the dexamethasone group (P<0.05, P<0.01); the α-SMA, TIMP-1, miRNA-146a, MMP-9 mRNA, and TIMP-1 mRNA expressions in high- and medium-dose WHD groups and the TGF-β1 expression in high-dose WHD group were lower than those in low-dose WHD group (P<0.05, P<0.01). The α-SMA, TIMP-1, miRNA-146a, and TIMP-1 mRNA expressions in the high-dose WHD group were lower than those in medium-dose WHD group (P<0.05, P<0.01). Pearson correlation analysis showed that there was a strong positive correlation between miRNA-146a and MMP-9 mRNA, TIMP-1 mRNA in the lung tissue of rats (r=0.956, 0.973, P<0.01). Conclusion WHD can effectively improve airway remodeling in asthmatic rats, which may be related to its inhibition of miRNA-146a expression and reduction of MMP-9, TIMP-1, mRNA, α-SMA and TGF-β1 expressions. |
| Key words: Wuhu Decoction young asthmatic rats miRNA-146a airway remodeling matrix metalloproteinase-9 tissue inhibitor of matrix metalloproteinases-1 α-smooth muscle actin transforming growth factor-β1 |
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