雷公藤甲素调控NF-κB/Twist 1信号通路抑制肺泡上皮细胞间质转分化的机制  

Analysis on the inhibitory effect of triptolide on NF-κB/Twist 1 signaling pathway on interstitial differentiation of alveolar epithelial cells

作　　者：陈宏[1] 陈群[2] 李全[1] CHEN Hong;CHEN Qun;LI Quan(First Affiliated Hospital,Heilongjiang University of Chinese Medicine,Harbin 150040,China;Acheng People's Hospital,Harbin 150300,China)

机构地区：[1]黑龙江中医药大学附属第一医院,哈尔滨150040 [2]哈尔滨市阿城区人民医院,哈尔滨150300

出　　处：《中华中医药杂志》2022年第3期1384-1388,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金面上项目(No.81573863,No.81774197);黑龙江省博士后资助项目(No.LBH-Z15213);中国博士后科学基金第13批特别资助(No.2020T130178);黑龙江省中医药科研项目(No.ZHY19-004)。

摘　　要：目的:探讨雷公藤甲素(TPL)基于核转录因子κB(NF-κB)/Twist1信号通路对抑制Ⅱ型肺泡上皮细胞间质转分化(EMT)而产生抗肺纤维化的作用机制。方法:40只SPF级昆明小鼠随机分为对照组,模型组,TPL低、高剂量组,每组10只。气管注射博来霉素诱导肺纤维化大鼠模型,造模后连续灌胃给药28 d。分别采用qPCR和(或)Western blot法检测肺组织和细胞内E钙黏蛋白(E-cadherin)、紧密连接蛋白-1(ZO-1)、波形蛋白(vimentin)、α-平滑肌肌动蛋白(α-SMA)、核转录因子κB抑制因子α(IκBα)、NF-κB p65、磷酸化NF-κB p65(p-p65)和Twist 1 mRNA及蛋白表达。结果:HE染色示TPL低、高剂量组成纤维细胞减少,增殖减弱,肺泡炎症细胞浸润程度以及肺纤维化病变程度均显著降低。与对照组比较,模型组肺组织ZO-1和E-cadherin的表达显著降低,α-SMA和vimentin的表达显著升高(P<0.05);与模型组比较,TPL高、低剂量组肺组织ZO-1和E-cadherin的表达显著升高,α-SMA和vimentin的表达显著降低(P<0.05);与TPL低剂量组比较,TPL高剂量组肺组织ZO-1和E-cadherin的表达显著升高,α-SMA和vimentin的表达显著降低(P<0.05)。与对照组比较,模型组肺组织p-IκBα、p-p65、NF-κB p65和Twist 1的表达显著升高(P<0.05);与模型组比较,TPL高、低剂量组肺组织p-IκBα、p-p65、NF-κB p65和Twist 1的表达显著降低(P<0.05),与TPL低剂量组比较,TPL高剂量组肺组织p-IκBα、p-p65、NF-κB p65和Twist 1的表达显著降低(P<0.05)。结论:TPL能够逆转肺泡Ⅱ型上皮细胞EMT、缓解肺纤维化,其机制可能与其降低细胞内IκBα磷酸化水平、抑制NF-κB p65的磷酸化和其核转移,进而下调Twist 1的表达有关。Objective: To discuss triptolide(TPL) inhibits epithelial mesenchymal transformation(EMT) of type II pulmonary epithelial cells based on nuclear transcription factor κB(NF-κB)/Twist 1 signaling pathway. Methods: A total of 40 SPF KM mice were divided into control group, model group, TPL low-dose group and TPL high-dose group, with 10 mice in each group.The rat model of pulmonary fibrosis was induced by intratracheal injection of BLM, after modeling, the drug was administered intragastric continuously for 28 days. Lung tissue and intracellular E-cadherin, tight junction protein-1(ZO-1), vimentin, α-smooth muscle actin(α-SMA), nuclear transcription factor κB inhibitory factor α(IκBα), NF-κB p65, phosphorylated NF-κB p65(pp65) and Twist 1 mRNA and protein expression were detected by qPCR and/or Western blot, respectively. Results: In TPL lowdose group and TPL high-dose group, ibroblasts were reduced and proliferation decreased through cell morphology observation.The degree of alveolar inflammatory cell infiltration and the degree of pulmonary fibrosis were significantly reduced. Compared with the control group, the expressions of ZO-1 and E-cadherin in lung tissues of model group were significantly decreased, and the expressions of α-SMA and vimentin were significantly increased(P<0.05). Compared with the model group, the expressions of ZO-1 and E-cadherin in lung tissues of TPL high-dose and low-dose groups were significantly increased, the expression of α-SMA and vimentin was significantly decreased(P<0.05), and the expression of ZO-1 and E-cadherin was significantly increased and the expression of α-SMA and vimentin was significantly decreased in TPL high-dose group compared with TPL low-dose group(P<0.05). Compared with the control group, the expression of p-I κBα, p-p65, NF-κB p65 and Twist 1 in lung tissues of model group were significantly increased(P<0.05). The expression of p-I κBα, p-p65, NF-κB p65 and Twist 1 in lung tissues of TPL high-dose and low-dose groups were significantly decrea

关 键 词：雷公藤甲素 核转录因子ΚB 肺纤维化 上皮细胞-间充质转化 Α-平滑肌肌动蛋白 

分 类 号：R285[医药卫生—中药学]