摘要
目的探讨麦味养肺汤(麦冬、五味子、党参等)通过调控p53/p21信号通路抑制Ⅱ型肺泡上皮细胞(AECⅡ)衰老抗肺纤维化的作用机制。方法将C57BL/6小鼠随机分为空白组、模型组、麦味养肺汤组(60 g·kg^(-1)·d^(-1))和阳性对照组(吡非尼酮,0.3 g·kg^(-1)·d^(-1)),每组6只。采用气管滴注博来霉素(5 mg·kg-1)复制肺纤维化小鼠模型;造模后次日开始灌胃给药,每日1次,连续给药21 d。检测并记录小鼠肺顺应性与肺阻力的变化;活体micro-CT成像法检测小鼠肺组织纤维化程度;采用HE、Masson染色法观察、评估肺组织炎症和纤维化程度;免疫组织化学法测定肺组织中Ⅰ/Ⅲ型胶原的表达;β-半乳糖苷酶(SA-β-gal)染色法测定肺组织细胞衰老情况;ELISA法测定肺组织中羟脯氨酸(Hyp)的含量;Western Blot法测定肺组织中Fibronectin、Vimentin、α-SMA、p53、p21、p16、SP-C蛋白表达;免疫荧光法检测小鼠肺组织中p21、p16及SP-C的表达。结果与空白组比较,模型组小鼠体质量显著下降(P<0.01),肺组织中炎症及纤维化评分均显著升高(P<0.01);肺顺应性显著下降(P<0.01),肺阻力显著增加(P<0.01);肺组织中Ⅰ/Ⅲ型胶原沉积量及Hyp含量显著增加(P<0.01);肺组织中Fibronectin、Vimentin、α-SMA蛋白表达显著上调(P<0.01);肺组织呈大面积SA-β-gal蓝染,细胞衰老程度明显,肺组织中p53、p21、p16等衰老相关蛋白表达显著上调(P<0.01);肺组织中p21、p16的红色荧光表达明显增强,Merge图像显示p21、p16与SP-C共染的橙色荧光明显增多,SP-C蛋白表达显著下调(P<0.01)。与模型组比较,麦味养肺汤组小鼠的体质量明显升高(P<0.05),肺组织中炎症及纤维化评分均明显降低(P<0.05,P<0.01);肺顺应性明显上升(P<0.05),肺阻力显著下降(P<0.01);肺组织中Ⅰ/Ⅲ型胶原沉积量及Hyp含量显著减少(P<0.01);肺组织中Fibronectin、Vimentin、α-SMA蛋白表达明显下调(P<0.05);肺组织SA-β-gal蓝染明显减轻,�
Objective To observe the mechanism of Maiwei Yangfei Decoction in inhibiting anti-pulmonary fibrosis of alveolar epithelial cell(AEC)Ⅱsenescence by regulating p53/p21 signaling pathway.Methods C57BL/6 mice were randomly divided into blank group,model group,Maiwei Yangfei Decoction group(60 g·kg^(-1)·d^(-1))and positive control group(Pirfenidone,0.3 g·kg^(-1)·d^(-1)),with 6 mice in each group.The mouse model of pulmonary fibrosis was replicated by intratracheal instillation of Bleomycin(BLM)(5 mg·kg-1).The next day after the establishment of the model,the drug was given by intragastric administration once a day for 21 consecutive days.The changes in lung compliance and pulmonary resistance were measured and recorded;the extent of pulmonary fibrosis was measured by in vivo micro-CT imaging;the extent of inflammation and fibrosis was observed and assessed by HE and Masson staining;the expression of type I/III collagen in lung tissue was determined by immunohistochemistry;the senescence of lung cells was determined byβ-galactosidase(SA-β-gal)staining;the content of hydroxyproline(Hyp)in lung tissue was determined by ELISA;the protein expressions of Fibronectin,Vimentin,α-SMA,p53,p21,p16 in lung tissue were determined by Western Blot;protein expressions of p53,p21,p16 and SP-C in lung tissue were detected by Western Blot;expressions of p21,p16 and SP-C in mouse lung tissue were detected by immunofluorescence.Results Compared with the blank group,the body mass of the model group was significantly decreased(P<0.01),the degree of inflammation and collagen deposition in the lung tissue were significantly increased(P<0.01),the pulmonary compliance was significantly decreased(P<0.01),the lung resistance was significantly increased(P<0.01),the deposition of typeⅠ/Ⅲcollagen and the content of Hyp in the lung tissue were significantly increased(P<0.01),and the protein expressions of Fibronectin,Vimentin andα-SMA in the lung tissue were significantly up-regulated(P<0.01).The lung tissue showed large area SA-β-ga
作者
王单
徐泳
黄同杏
韩迪
王晶
魏云
周贤梅
WANG Dan;XU Yong;HUANG Tongxing;HAN Di;WANG Jing;WEI Yun;ZHOU Xianmei(Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029 Jiangsu,China;School of Traditional Chinese Medicine,School of Integrated Chinese and Western Medicine,Nanjing University of Chinese Medicine,Nanjing 210029 Jiangsu,China)
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2023年第6期754-763,共10页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
国家自然科学基金面上项目(82074358)
