文献类型：期刊文献

中文题名：基于RhoA/ROCK通路探讨宣肺化浊方对急性肺损伤小鼠肺内皮细胞微管解聚的影响及机制研究

英文题名：Study on the effects and mechanism of Xuanfei Huazhuo Formula on microtubule depolymerization of pulmonary endothelial cells in mice with acute lung injury based on RhoA/ROCK pathway

作者：朱鹏[1];雍文兴[2];宋忠阳[2];吴亚娜[1];张录梅[1];徐倩[1];李爱玉[1];郝国雄[1];刘瑜[1];张志明[3]

第一作者：朱鹏

机构：[1]甘肃中医药大学,兰州730000;[2]甘肃中医药大学附属医院,兰州730000;[3]甘肃省中医院,兰州730050

第一机构：甘肃中医药大学

年份：2023

卷号：38

期号：7

起止页码：3323

中文期刊名：中华中医药杂志

外文期刊名：China Journal of Traditional Chinese Medicine and Pharmacy

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD2023_2024】；

基金：国家中医药管理局科研项目(No.2021ZYLCYJ08);高端人才承担省级科技计划项目(“长江学者奖励计划”特聘教授)(No.甘科计[2022]20号-9);甘肃省科技计划重大专项(No.22ZD1FA001);兰州市人才创新创业项目(No.2021-RC-80);甘肃省中医药管理局科研项目(No.GZKP-2021-15)。

语种：中文

中文关键词：急性肺损伤;RhoA/ROCK通路;宣肺化浊方;内皮细胞;通透性

外文关键词：Acute lung injury(ALI);RhoA/ROCK pathway;Xuanfei Huazhuo Formula;Endothelial cells;Permeability

摘要：目的:基于RhoA/ROCK通路探讨宣肺化浊方对急性肺损伤(ALI)小鼠肺微血管内皮细胞骨架肌动蛋白微管的调控作用及机制研究。方法:72只雄性C57BL/6小鼠按体质量随机分为空白组,模型组,盐酸法舒地尔组(10 mg/kg)及宣肺化浊方低、中、高剂量组(0.06、0.11、0.22 mg/kg)。除空白组外,其余各组采用腹腔注射LPS建立小鼠ALI模型。计算BALF中细胞总数;HE染色观察肺组织病理学变化;透射电镜观察肺血管内皮细胞线粒体的超微结构;RT-PCR法检测肺组织中通路相关因子RhoA、ROCK mRNA表达量,Western Blot及免疫组化法检测肺组织中微管相关蛋白p-Tau、p-MAP-4、Ace-tubulin的表达量。结果:与模型组比较,各治疗组小鼠肺组织BALF中细胞总数均显著降低(P<0.01);宣肺化浊方中高剂量组、阳性对照组肺间质炎性细胞浸润,肺间质水肿,肺泡壁增厚,肺泡腔破坏程度均比模型组明显减轻;宣肺化浊方中、高剂量组及阳性对照组:线粒体形态结构较模型及低剂量组好,线粒体嵴均轻度模糊;与模型组比较,宣肺化浊方中、高剂量组及阳性对照组RhoA、ROCK mRNA水平显著降低(P<0.01);与模型组比较,各治疗组小鼠肺组织P-Tau、P-MAP-4蛋白表达水平显著降低(P<0.01),Ace-tubulin蛋白表达水平显著升高(P<0.05)。结论:宣肺化浊方可通过调控RhoA/ROCK通路来更好地抑制微管解聚从而降低LPS所致ALI小鼠肺微血管内皮细胞的通透性,对肺组织起到保护作用。
Objective:To investigate the regulatory effect and mechanism of Xuanfei Huazhuo Formula on actin microtubules in pulmonary microvascular endothelial cells of ALI mice based on RhoA/ROCK pathway.Methods:A total of 72 male C57BL/6 mice were randomly divided into blank group,model group,fasudil hydrochloride group(10 mg/kg),and low,medium,and high dose Xuanfei Huazhuo Formula groups(0.06,0.11,0.22 mg/kg)according to their body mass.Except for the blank group,other groups were injected intraperitoneally with LPS to establish mouse ALI models.Calculate the total number of cells in BALF;HE staining was used to observe the pathological changes of lung tissue;The ultrastructure of mitochondria in pulmonary vascular endothelial cells was observed by transmission electron microscopy;RT-PCR was used to detect the mRNA expression of pathway related factors RhoA and ROCK in lung tissue,and Western Blot and immunohistochemistry were used to detect the expression of microtubule related proteins p-Tau,p-MAP-4,and Ace-tubulin in lung tissue.Results:Compared with the model group,the total number of cells in BALF of mouse lung tissue in each treatment group significantly decreased(P<0.01).In the middle and high dose group of Xuanfei Huazhuo Formula and the positive control group,the infiltration of inflammatory cells in the pulmonary interstitium,pulmonary interstitial edema,thickening of the alveolar wall,and destruction of the alveolar cavity were significantly reduced compared to the model group;The middle and high dose groups of Xuanfei Huazhuo Formula and the positive control group showed that the mitochondrial morphology and structure were better than those of the model and low dose groups,and the mitochondrial cristae were slightly blurred;Compared with the model group,the levels of RhoA and ROCK mRNA in the middle and high dose groups of Xuanfei Huazhuo Formula and the positive control group were significantly lower(P<0.01).Compared with the model group,the expression levels of P-Tau and P-MAP-4 protiens in the mouse lung tissue in each medication groups were significantly reduced(P<0.01),while the expression levels of Ace-tubulin protein were significantly increased(P<0.05).Conclusion:Xuanfei Huazhuo Formula can better inhibit microtubule depolymerization by regulating RhoA/ROCK pathway,thereby reducing the permeability of pulmonary microvascular endothelial cells in ALI mice induced by LPS,and has a protective effect on lung tissue.