鱼腥草素钠通过PI3K/Akt/mTOR信号通路缓解百草枯诱导的肺纤维化  

Sodium houttuyfonate attenuates paraquat induced pulmonary fibrosis by regulating the PI3K/Akt/mTOR signaling pathway

作　　者：王龙海 何青云 董双虎 WANG Longhai;HE Qingyun;DONG Shuanghu(Department of Intensive Care Unit,Jiangxi Province Hospital of Integrated Traditional Chinese andWestern Medicine,Nanchang,Jiangxi,330003,China)

机构地区：[1]江西省中西医结合医院重症医学科,江西南昌330003

出　　处：《当代医学》2023年第22期7-13,共7页Contemporary Medicine

基　　金：江西省卫生健康委科技计划(202140056)。

摘　　要：目的探讨鱼腥草素钠(SH)对百草枯(PQ)导致的肺纤维化的缓解作用及其潜在作用机制。方法选取5周龄雄性SD大鼠45只,随机分为对照组、PQ组与SH组,每组15只。PQ组大鼠灌胃40 mg/kg的PQ溶液,SH组在灌胃40 mg/kg的PQ溶液1 h后灌胃100 mg/kg的SH,对照组灌胃等量0.9%氯化钠溶液;采用Szapiel法分析肺组织的肺炎和肺纤维化程度;酶联免疫吸附(ELISA)法检测肺组织中超氧歧化酶(SOD)、丙二醛(MDA)和羟脯氨酸(HYP)的含量;蛋白印迹法(Westernblot)测定肺组织中α-平滑肌肌动蛋白(α-SMA)、转化生长因子-β1(TGF-β1)和E-钙黏蛋白(E-cadherin)的含量;不同浓度(1、10、20、30、40μmol/L)SH联合300μmol/L的PQ作用体外人肺泡上皮细胞(HPAEpiC)后,采用细胞计数试剂-8(CCK-8)法评价HPAEpiC细胞的活力;ELISA法检测HPAEpiC细胞中白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)的含量;Westernblot测定HPAEpiC细胞中α-SMA、TGF-β1、E-cadherin、磷酸化丝苏氨酸蛋白激酶(p-Akt)、磷酸化雷帕霉素靶蛋白(p-mTOR)和磷酸化磷酸肌醇3激酶(p-PI3K)蛋白的含量。结果SH组肺炎和肺纤维化程度显著低于PQ组(P<0.05);PQ组SOD水平明显低于对照组,MDA和HYP水平明显高于对照组(P<0.05);SH组SOD水平明显高于PQ组,MDA和HYP水平明显低于PQ组,差异有统计学意义(P<0.05)。PQ组肺组织中α-SMA和TGF-β1蛋白的表达水平高于对照组(P<0.05),而E-cadherin的蛋白表达水平明显低于对照组(P<0.05);SH组α-SMA和TGF-β1蛋白水平明显低于PQ组,E-cadherin水平明显高于PQ组,差异有统计学意义(P<0.05)。SH可有效缓解PQ诱导的HPAEpiC细胞活力下降,SH最适浓度为10μmol/L。PQ组IL-1β、IL-6、TNF-α水平明显高于对照组(P<0.05);SH组IL-1β、IL-6和TNF-α水平明显低于PQ组(P<0.05)。PQ组HPAEpiC细胞中α-SMA和TGF-β1蛋白表达水平明显高于对照组,E-cadherin蛋白表达水平明显低于对照组(P<0.05);SH组HPAEpiC细胞中α-SMA和Objective The purpose of the present study was to investigate the protective effect and underlying mechanism of sodium houttuyfo-nate(SH)on paraquat(PQ)poisoning induced pulmonary fibrosis.Methods 45 cases 5-week-old male SD rats were selected,and they were ran-domly divided into the control group,PQ group and SH group,with 15 cases in each group.Rats in PQ group were given 40 mg/kg PQ by gavage,rats in SH group were given 100 mg/kg SH after the gavage of 40 mg/kg PQ for 1 h,rats in control group were given the same amount of 0.9%sodi-um chloride solution by intragastric administration.Szapiel's method was utilized to analyze the degree of alveolitis and pulmonary fibrosis in lung tissue;ELISA method was employed to analyze the levels of superoxide dismutase(SOD),malondialdehyde(MDA),and hydroxyproline(HYP)in lung tissue;Western blot was utilized to evaluate the expression ofα-smooth muscle actin(α-SMA),transforming growth factor-β(TGF-β1)and E-cad-herin in lung tissue;human alveolar epithelial cells(HPAEpiC)were incubated with SH(1,10,20,30,40μmol/L)combined with PQ(300μmol/L);cell counting kit-8(CCK-8)assay was employed to evaluate the viability of HPAEpiC cells.ELISA was utilized to detect the levels of interleukin-1β(IL-1β),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in HPAEpiC cells.Western blot was utilized to evaluate the expression ofα-SMA,TGF-β1,E-cadherin,p-seronine protein kinase(p-Akt),p-target protein of rapamycin(p-mTOR)and p-inositol 3 kinase(p-PI3K)in HPAEpiC cells.Results The degree of pneumonia and pulmonary fibrosis in the SH group was significantly lower than those in the PQ group(P<0.05).SOD lev-els in the PQ group was significantly lower than that in the control group,MDA and HYP levels were significantly higher than those in the control group(P<0.05);and the SOD level in the SH group was significantly higher than that in the PQ group,MDA and HYP levels were significantly low-er than the PQ group,and the differences were statistically significant(P<0.05).The expression l

关 键 词：鱼腥草素钠 肺纤维化 百草枯 磷酸肌醇3激酶 

分 类 号：R28[医药卫生—中药学]