余甘子醇提物对硅肺小鼠的保护作用及与Nrf2/ARE信号通路的相关性  

Protective Effect of Alcohol Extract of Phyllanthi Fructus on Silicosis Mice and Its Correlation with Nrf2/ARE Signaling Pathway

作　　者：张玉蝶 李欣[1] 何晓艳 吴丽娟[1] 余蓉[1] 杨培富 陈大义[1] ZHANG Yudie;LI Xin;HE Xiaoyan;WU Lijuan;YU Rong;YANG Peifu;CHEN Dayi(Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;Liangshan Zhoushengsen Biological Technology Co.Ltd.,Xichang 615000,China)

机构地区：[1]成都中医药大学,成都611137 [2]凉山州晟森生物科技有限公司,四川西昌615000

出　　处：《中国实验方剂学杂志》2023年第9期129-136,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(82004251);凉山州农业科技创新项目(19NYCX0022)。

摘　　要：目的:探讨余甘子醇提物对二氧化硅(SiO2)诱导的硅肺小鼠的影响及可能的作用机制。方法:将SPF级雄性昆明小鼠36只随机分为空白组、模型组、余甘子高剂量组(800 mg·kg^(-1))、余甘子中剂量组(400 mg·kg^(-1))、余甘子低剂量组(200 mg·kg^(-1))、汉防己甲素组(0.039 mg·kg^(-1)),每组6只。除空白组以外,其余组均采用静式染毒法复制硅肺模型。灌胃28 d后取出肺组织计算脏器系数;采用苏木素-伊红(HE)染色和Masson染色检测小鼠肺组织的组织形态;采用酶联免疫吸附测定法(ELISA)检测血清中的羟脯氨酸(HYP)、超氧化物歧化酶(SOD)、丙二醛(MDA)、过氧化氢酶(CAT)的含量;应用蛋白免疫印迹法(Western blot)和实时荧光定量聚合酶链式反应(Real-time PCR)技术检测核因子E2相关因子2(Nrf2)、血红素氧合酶-1(HO-1)、依赖型辅酶还原酶/醌氧化还原酶1(NQO1)、Kelch样环氧氯丙烷相关蛋白1(Keap1)蛋白和mRNA的表达。结果:与空白组比较,模型组小鼠肺组织形态结构严重损伤,炎性细胞浸润,纤维组织增生;血清中SOD和CAT的含量均显著减少(P<0.01),HYP和MDA的含量均显著增加(P<0.01);肺组织中Nrf2、HO-1、NQO1蛋白与mRNA的表达均显著减少(P<0.01),Keap1蛋白与mRNA的表达均明显增加(P<0.05,P<0.01)。与模型组比较,余甘子高、中剂量组小鼠的肺组织形态结构得到明显恢复,胶原沉积减少;余甘子高、中剂量组血清的SOD和CAT含量均明显增加(P<0.05,P<0.01),HYP和MDA含量均显著减少(P<0.01);余甘子高、中剂量组肺组织的Nrf2、HO-1、NQO1蛋白与mRNA的表达均明显增加(P<0.05,P<0.01),Keap1蛋白与mRNA的表达均明显减少(P<0.05,P<0.01)。结论:余甘子醇提物可以抑制硅肺小鼠肺纤维化,其机制可能与Nrf2/抗氧化反应元件(ARE)信号通路的调控有关。Objective:To explore the effect and underlying mechanism of alcohol extract of Phyllanthi Fructus on silicosis mice induced by silicon dioxide(SiO2).Method:Thirty-six male Kunming mice of SPF grade were randomly divided into a blank group,a model group,high-,medium,and low-dose Phyllanthi Fructus groups(800,400,200 mg·kg^(-1)),and a tetrandrine group(0.039 mg·kg^(-1)),with six mice in each group.The silicosis model was induced by static SiO2 exposure in mice except for those in the blank group.After 28 days of administration by gavage,the lung tissues were collected and the organ coefficient was calculated.Hematoxylin-eosin(HE)staining and Masson staining were used to detect the morphology of lung tissues.The content of hydroxyproline(HYP),superoxide dismutase(SOD),malondialdehyde(MDA),and catalase(CAT)in serum was detected by enzyme-linked immunosorbent assay(ELISA).Western blot and Real-time polymerase chain reaction(Real-time PCR)were used to detect the protein and mRNA expression of nuclear factor E2-related factor 2(Nrf2),heme oxygenase-1(HO-1),NAD(P)H:quinone oxidoreductase 1(NQO1),and Kelch-like ECH-associated protein 1(Keap1),respectively.Result:Compared with the blank group,the model group showed seriously damaged morphological structure of lung tissues with inflammatory cell infiltration and fibrous tissue proliferation,reduced serum content of SOD and CAT(P<0.01),increased content of HYP and MDA(P<0.01),down-regulated protein and mRNA expression of Nrf2,HO-1,and NQO1(P<0.01),and up-regulated protein and mRNA expression of Keap1(P<0.05,P<0.01).Compared with the model group,the high-and medium-dose Phyllanthi Fructus groups showed significantly restored morphological structure of lung tissues with reduced collagen deposition,increased serum content of SOD and CAT(P<0.05,P<0.01),decreased content of HYP and MDA(P<0.01),up-regulated protein and mRNA expression of Nrf2,HO-1,and NQO1(P<0.05,P<0.01),and down-regulated protein and mRNA expression of Keap1(P<0.05,P<0.01).Conclusion:The alcohol extract of Phyl

关 键 词：余甘子 硅肺 抗纤维化 抗氧化 核因子E2相关因子2(Nrf2)/抗氧化反应元件(ARE)信号通路 

分 类 号：R242[医药卫生—中医临床基础] R2-0[医药卫生—中医学] R2-031R285R825.6R56