文献类型：期刊文献

中文题名：镰形棘豆总黄酮通过调控JAK1/STAT1/SOCS3信号通路减轻特发性肺纤维化

英文题名：Total flavonoids of Oxytropis falcata Bunge attenuate idiopathic pulmo?nary fibrosis by regulating JAK1/STAT1/SOCS3 signaling pathway

作者：李欣泽[1,2,3];王彦君[1,2,3];李杨[1];梁乾坤[1];陈彦文[1];杨玲玲[1];明海霞[1,2]

第一作者：李欣泽

机构：[1]甘肃中医药大学基础医学院,甘肃兰州730000;[2]甘肃省高校重大疾病分子医学与中医药防治研究重点实验室,甘肃兰州730000;[3]甘肃中医药大学中西医结合研究所,甘肃兰州730000

第一机构：甘肃中医药大学基础医学院（敦煌医学研究所）

年份：2022

卷号：38

期号：5

起止页码：905

中文期刊名：中国病理生理杂志

外文期刊名：Chinese Journal of Pathophysiology

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD2021_2022】；

基金：国家自然科学基金资助项目(No.81860830)。

语种：中文

中文关键词：镰形棘豆总黄酮;特发性肺纤维化;JAK1/STAT1/SOCS3信号通路

外文关键词：Total flavonoids of Oxytropis falcata Bunge;Idiopathic pulmonary fibrosis;JAK1/STAT1/SOCS3 signaling pathway

摘要：目的:探讨镰形棘豆总黄酮(FOFB)调控Janus激酶1(JAK1)/信号转导及转录激活蛋白1(STAT1)/细胞因子信号传送阻抑物3(SOCS3)信号通路干预特发性肺纤维化(IPF)体外模型的机制。方法:将50只SPF级大鼠随机分为空白血清组、含低浓度FOFB血清组、含中浓度FOFB血清组、含高浓度FOFB血清组和含吡非尼酮血清组,每组10只,并按照中药血清药理学方法提取含药血清。将人胚肺成纤维细胞系HFL-1分正常组、模型组、空白血清组、含低浓度FOFB血清组、含中浓度FOFB血清组、含高浓度FOFB血清组和含吡非尼酮血清组。采用纤维化诱导剂转化生长因子β1建立IPF体外模型,CCK-8法筛选含药血清最佳干预时间,进行最佳时间干预;RT-qPCR及Western blot检测Ⅰ型胶原(COL I)、Ⅲ型胶原(COL Ⅲ)和α-平滑肌肌动蛋白(α-SMA)的表达,以验证模型建立是否成功;透射电子显微镜观察FOFB对细胞内粗面内质网的影响;RT-qPCR检测SOCS3、肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、细胞间黏附分子1(ICAM1)和单核细胞趋化蛋白1(MCP1)的mRNA表达;Western blot检测p-JAK1、p-STAT1、SOCS3、TNF-α、IL-1β、ICAM1和MCP1的蛋白水平。结果:CCK-8法检测含药血清最佳干预时间为120 h。与正常组相比,模型组中COL I、COL Ⅲ和α-SMA的表达显著升高(P<0.01),提示模型建造成功;经过FOFB干预后,COL I、COL Ⅲ和α-SMA的表达显著下降(P<0.01)。透射电子显微镜结果显示,与正常组相比,模型组中粗面内质网数量增加;经过FOFB干预后,粗面内质网数量逐渐降低,且其结构逐渐呈囊泡状。RT-qPCR及Western blot结果显示,与正常组比较,p-JAK1、p-STAT1、TNF-α、IL-1β、ICAM1及MCP1的表达水平显著升高,SOCS3表达水平显著降低(P<0.01);经FOFB干预后,与模型组比较,JAK1/STAT1信号通路被显著抑制,其中p-JAK1、pSTAT1、TNF-α、IL-1β、ICAM1及MCP1的表达水平显著降低,SOCS3的表达水平显著升高(P<0.01)。结论:FOFB在IPF体外模型中可能通过升高SOCS3表达而抑制JAK1/STAT1信号通路,从而延缓IPF的进展。
AIM:To investigate the effect of total flavonoids of Oxytropis falcata Bunge(FOFB)on idiopathic pulmonary fibrosis(IPF)in vitro.METHODS:Fifty SPF rats were randomly divided into animal serum group,animal serum containing low-concentration FOFB group,animal serum containing medium-concentration FOFB group,animal serum containing high-concentration FOFB group and animal serum containing pirfenidone group,with 10 rats in each group. Drug-containing serum was extracted according to the method of serum pharmacology of traditional Chinese medicine. Human fetal lung fibroblasts(HFL-1)were divided into control group,model group,animal serum group,animal serum containing low-concentration FOFB group,animal serum containing medium-concentration FOFB group,animal serum containing high-concentration FOFB group and animal serum containing pirfenidone group. Transforming growth factor-β1 was used to establish IPF model in vitro. The optimal intervention time of drug-containing serum was selected by CCK-8 assay. The expression levels of collagen type Ⅰ(COL I),collagen type Ⅲ(COL Ⅲ)and α-smooth muscle actin(α-SMA)were detected by RT-qPCR and Western blot. The effect of FOFB on rough endoplasmic reticulum was observed by transmission electron microscopy. The mRNA expression of suppressor of cytokine signaling 3(SOCS3),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),intercellular adhesion molecule 1(ICAM1)and monocyte chemoattractant protein 1(MCP1)was detected by RT-qPCR. The protein levels of phosphorylated Janus kinase 1(p-JAK1),phosphorylated signal transducer and activator of transcription 1(p-STAT1),SOCS3,TNF-α,IL-1β,ICAM1 and MCP1 were detected by Western blot.RESULTS:The optimal intervention time was 120 h according to the results of CCK-8 assay.Compared with control group,the expression levels of COL I,COL Ⅲ and α-SMA in model group were significantly increased(P<0. 01),indicating that the IPF model was successfully constructed. After FOFB intervention,the expression levels of COL I,COL Ⅲ and α-SMA significantly decreased(P<0. 01). The results of transmission electron microscopy showed that the number of rough endoplasmic reticulum in model group was increased compared with conrtol group,while it was gradually decreased and the structure appeared as vesicles after FOFB intervention. Compared with control group,the expression levels of p-JAK1,p-STAT1,TNF-α,IL-1β,ICAM1 and MCP1 in model group were significantly increased,while the expression of SOCS3 was significantly decreased(P<0. 01). Compared with model group,the JAK1/STAT1 signaling pathway was significantly inhibited after FOFB intervention,the expression levels of p-STAT1,TNF-α,IL-1β,ICAM1 and MCP1 were significantly decreased,while the expression of SOCS3 was significantly increased(P<0. 01).CONCLUSION:The FOFB attenuate IPF by inhibiting JAK1/STAT1 signaling pathway and increasing SOCS3.