Idiopathic pulmonary fibrosis(IPF)is a chronic interstitial lung disease marked by the significant in-volvement of fibroblasts,macrophages and lymphocytes.The CD27/CD70 axis is pivotal in shaping the immune mi-croenvironment present in the fibrotic aeras of lungs.This mentioned interaction involves molecular pathways that work in tandem with lung immune cells,particularly exerting a suppressive influence in the early phases of pulmo-nary fibrosis.Consequently,the CD27/CD70 axis presents a promising new target for immunotherapy in IPF.

Secretory immunoglobulin A (SIgA) is one of the most abundant immunoglobulin subtypes among mucosa, which plays an indispensable role in the first-line protection against invading pathogens and antigens. Therefore, the role of respiratory SIgA in respiratory mucosal immune diseases has attracted more and more attention. Although the role of SIgA in intestinal mucosal immunity has been widely studied, the cell types responsible for SIgA and the interactions between cells are still unclear. Here, we conducted a wide search of relevant studies and sorted out the relationship between SIgA and some pulmonary diseases (COPD, asthma, tuberculosis, idiopathic pulmonary fibrosis, COVID-19, lung cancer), which found SIgA is involved in the pathogenesis and progression of various lung diseases, intending to provide new ideas for the prevention, diagnosis, and treatment of related lung diseases.

OBJECTIVE:Idiopathic pulmonary fibrosis (IPF) is considered as a chronic interstitial lung disease with underlying mechanism of IPF remaining unclear, while there are no definitive treatment options. In recent years, scientists have gradually paid attention to the influence of angiogenesis on IPF. Because IPF is a progressive with microvascular remodeling disorder, scientists have postulated that angiogenesis may also be one of the initiating and contributing factors of the disease. Bupleurum is a common natural Chinese herbal medicine with antibacterial, anti-inflammatory, anti-tumor and other pharmacological effects. As the most important active monomer of Bupleurum, Saikosaponin-d (SSd) is a new discovery with anti-pulmonary fibrosis (PF) activity. This study attempts to investigate the role of SSd in the interference of PF through regulation of angiogenesis in IPF through Angiopoietin (Angpt) /Tie receptor 2 (Tie2) pathway. METHODS:Randomly, we allocated C57BL/6 mice into four groups (n = 20 in each group). Afterwards, establishment of IPF model was accomplished via intratracheal administration of bleomycin (BLM, 5 mg/kg), while corresponding drug intervention was given accordingly. On 3rd, 7th, 14th and 28th days after modeling, we performed histopathological examination through staining. Meanwhile, immunohistochemistry (IHC) of PF and the expression of related factors were observed, while Ang/Tie2 pathway was assessed by ELISA with the effect of SSd on angiogenesis related proteins in IPF being explored with IHC and Western Blot technique. RESULTS:Our results showed that SSd could reduce inflammation and PF levels in lung tissue of experimental mice, while levels of angiogenesis-related factors, namely Tie-2, Ang-1 and ANGPT2 (Ang-2), fibrosis- associated factors like Alpha-smooth muscle actin (α-SMA), collagen-I and hydroxyproline in SSd and dexamethasone (DXM) mice were significantly reduced at each time point compared to BLM (p < 0.01). Additionally, we discovered substantial decreased expressions of Ang-1, Ang-2, Tie-2, α-SMA and collagen-I at protein level in SSd and DXM mice at each time point compared to BLM (p < 0.05). Besides, insignificant differences were observed between SSd and DXM groups (p > 0.05). CONCLUSION:This study has demonstrated that SSd could down-regulate the expression of ANG-1, Ang-2 and Tie2 in the Ang/Tie2 pathway, and may reduce lung inflammation and PF in BLM-induced mice via inhibition of angiogenesis.

背景:非编码RNA随着高通量技术的发展而逐渐引发关注,其广泛参与呼吸系统各疾病病程.特发性肺纤维化进程伴随着干细胞功能异常,近来研究表明非编码RNA与干细胞功能异常存在联系,进而影响肺纤维化进展.以非编码RNA为靶点的基因疗法暗示了阻断纤维化过程的可能性.目的:总结非编码RNA在特发性肺纤维化干细胞功能异常和治疗中的研究进展.方法:以"IPF,Stem cells,miRNA,lncRNA,circRNA,treatment"为英文检索词,以"特发性肺纤维化、干细胞、miRNA、lncRNA、circRNA、治疗"为中文检索词,检索PubMed、Wiley InterScience、中国知网和万方数据库,范围囊括近10年的文献,然后依据纳入和排除标准进行筛选,最终纳入82篇文献进行分析.结果 与结论:①文章首次总结了近年来非编码RNA在特发性肺纤维化的干细胞异常机制中的相关性研究,揭示了miRNA,lncRNA及circRNA共同调控了干细胞的异常激活、分化等活动,并且miRNA常作为后两者作用的中心环节.②现有的临床试验一定程度上肯定了干细胞移植疗法治疗特发性肺纤维化的安全性,但在疗效方面,仅稍改善了患者弥散功能却未能明确缓解纤维化病变的程度,其结果还需要更多设立安慰剂对照的随机试验来验证.③靶向非编码RNA的基因治疗优势在于具有多基因调控特性,可以从上游干预干细胞异常,目前其在体内外实验已取得突破性进展,并被证实与部分药物的作用靶点相关.④然而,特发性肺纤维化基因疗法的缺点是只能产生短期效应,同时其面临着有效剂量、体内稳定性及输送等问题,这些问题还需未来研究进一步解决.⑤总之,虽然非编码RNA疗法自身存在缺陷,但也不失为一种改善肺纤维化干细胞功能异常的新途径,将其与干细胞移植结合,取长补短,可能发挥更好的疗效.

目的 研究柴胡皂甙d(SSd)对特发性肺纤维化(IPF)中Ⅱ型肺泡上皮细胞(AT-Ⅱ)自噬的调控作用及其对纤维化程度的干预作用.方法 转化生长因子β1(TGF-β1,5 ng/mL)诱导A549细胞48 h建立肺纤维化细胞模型,分别联合SSd低、中、高浓度(2.5、5、10 μg/mL)、雷帕霉素(100 ng/mL)处理细胞.CCK8法检测各组细胞存活率,Western Blot法检测E-钙黏蛋白(E-cadherin)、Ⅰ型胶原蛋白(Col-Ⅰ)、α-平滑肌肌动蛋白(α-SMA)、Beclin1、微管相关蛋白1轻链3B-Ⅱ(LC3B-Ⅱ)/LC3B-Ⅰ、p62、哺乳动物雷帕霉素靶蛋白(mTOR)通路活化蛋白表达水平,单丹磺酰尸胺(MDC)染色法检测细胞内自噬泡形成.结果 与正常组比较,模型组细胞存活率下降(P＜0.05),E-cadherin、Beclin1、LC3B-Ⅱ/LC3B-Ⅰ蛋白表达水平亦降低(P＜0.05,P＜0.01),Col-Ⅰ、α-SMA、p62、mTOR通路活性蛋白表达水平升高(P＜0.05,P＜0.01),胞质内荧光标记的自噬泡较少.与模型组比较,SSd高浓度组细胞存活率升高(P＜0.05),SSd 组及雷帕霉素组 E-cadherin、Beclin1、LC3B-Ⅱ/LC3B-Ⅰ蛋白表达水平升高(P＜0.05,P＜0.01),Col-Ⅰ、α-SMA、p62蛋白表达水平降低(P＜0.05,P＜0.01),SSd高浓度组及雷帕霉素组p-mTOR/mTOR、p-AKT1/AKT1、p-S6K1/S6K1表达水平降低(P＜0.05,P＜0.01),SSd高浓度组及雷帕霉素组细胞质内荧光标记的自噬泡增多.与SSd低、中低浓度组比较,SSd高浓度组及雷帕霉素组Ecadherin、LC3B-Ⅱ/LC3B-Ⅰ表达水平升高(P＜0.05,P＜0.01),Col-Ⅰ、α-SMA、p62表达水平降低(P＜0.05,P＜0.01).结论 SSd可改善TGF-β1诱导的AT Ⅱ纤维化改变,可能通过抑制mTOR通路激活诱导细胞自噬发挥作用.

目的 探索circ_0007762和miR-18a-5p的相互作用及在肺成纤维细胞中的作用.方法 利用生物信息学分析circ_0007762在特发性肺纤维化(IPF)患者的表达并筛选其miRNA靶标,在转化生长因子(TGF)-β1干预的肺成纤维细胞HFL1中验证其表达.双荧光素酶报告基因实验验证miR-18a-5p mimics与报告基因载体共转染后的荧光素酶活性.CCK-8法检测circ_0007762、miR-18a-5p敲低及TGF-β1、自噬抑制剂3-甲基腺嘌呤(3-MA)干预后的细胞活力.Western blot检测circ_0007762、miR-18a-5p敲低及TGF-β1、3-MA干预后的α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原(collagenⅠ)、P62及微管相关蛋白1轻链3β(LC3B)水平.结果 HFL1细胞中,TGF-β1组较Control组circ_0007762表达水平上调,miR-18a-5p表达下调(P<0.05).在circ_0007762野生型载体中,过表达miR-18a-5p抑制了荧光素酶活性(P<0.05).circ_0007762敲低后细胞增殖活力下降,并由miR-18a-5p的抑制而恢复,同时3-MA可逆转TGF-β1诱导的成纤维细胞增殖(P<0.05).TGF-β1促进α-SMA、collagenⅠ及LC3BⅡ/Ⅰ表达,而抑制P62水平(P<0.05).相较于TGF-β1+si-NC组,TGF-β1+si-circ_0007762组P62表达上调,其余蛋白下调,并能由miR-18a-5p的抑制而逆转(P<0.05).此外,3-MA增强了P62的表达而降低了α-SMA、collagenⅠ的表达及LC3BⅡ/Ⅰ水平(P<0.05).结论 circ_0007762可与miR-18a-5p相互作用,通过激活自噬促进肺成纤维细胞增殖,诱导纤维化相关表型.

目的:探讨N-乙酰半胱氨酸(N-acetylcysteine,NAC)对哮喘模型小鼠气道炎症和氧化应激反应的影响及可能的机制.方法:选择18只SPF级BALB/c雌性小鼠,随机均分为对照组、哮喘组、N-乙酰半胱氨酸组,每组6只.哮喘组、N-乙酰半胱氨酸组雾化激发构建小鼠哮喘模型,N-乙酰半胱氨酸组每次雾化开始前30 min灌胃给药N-乙酰半胱氨酸(300 mg/kg),对照组予等量PBS处理,末次激发后24 h内取检测标本.收集支气管肺泡灌洗液行炎症细胞计数;ELISA法检测血清中细胞因子IL-5、IL-13和IFN-γ含量;取小鼠肺组织行HE染色观察病理变化并行炎症评分;比色法测定肺组织中丙二醛和谷胱甘肽含量;实时荧光定量PCR测肺组织核因子E2相关因子2(nu-clear factor erythroid 2 related factor 2,Nrf2)、血红素加氧酶-1(heme oxygenase-1,HO-1)mRNA表达水平.结果:与对照组相比,哮喘组肺组织大量炎性细胞浸润、肺泡结构破坏明显,炎症细胞数、IL-5、IL-13含量、炎症评分、丙二醛含量均明显升高(P均<0.05),IFN-γ和谷胱甘肽含量均明显降低(P均<0.05),Nrf2和HO-1 mRNA表达差异无统计学意义(P>0.05);与哮喘组相比,N-乙酰半胱氨酸组肺组织炎症细胞浸润减少、肺泡结构破坏减轻,炎症细胞数、IL-5和IL-13含量、炎症评分、丙二醛含量均明显降低(P均<0.05),IFN-γ、谷胱甘肽含量和Nrf2、HO-1 mRNA表达均明显升高(P均<0.05).结论:NAC可减轻哮喘小鼠气道炎症及氧化应激反应,可能通过激活Nrf2/HO-1信号通路发挥保护作用.

细胞焦亡又称细胞炎症性坏死，表现为细胞不断胀大直至细胞膜破裂释放细胞内容物及促炎因子如IL-1β、IL-18，从而引起激烈的炎症反应，是一种程序性细胞死亡方式。细胞焦亡主要由半胱天冬氨酸酶（caspase）介导，下游多种细胞焦亡蛋白（gasdermin）家族成员发生剪切和多聚化，导致细胞穿孔进而引起细胞死亡。越来越多的研究表明，细胞焦亡参与了间质性肺疾病的发生、发展过程。本文主要就细胞焦亡的发展历程、信号通路、与其他程序性细胞死亡方式的对比及其与间质性肺疾病之间关系的相关研究进行综述，为研究间质性肺疾病的发病机制、诊断及治疗提供参考。

Idiopathic pulmonary fibrosis (IPF) is an age-related chronic progressive fibrotic interstitial lung disease of unknown reasons and irreversibility.Mitophagy is a process, in which cells selectively remove abnormal mitochondria through autophagy mechanism.Abnormal mitophagy was found in IPF alveolar epithelial cells, fibroblasts and macrophages, which can promote low elasticity in lung tissue and increase sensitivity to fibrosis response activation.This article reviews the mechanism of mitochondrial autophagy in IPF lung cells.

Eosinophil extracellular trap (EET) is composed of a reticular complex of DNA fibers and granular proteins.Recent researches have suggested that EET can not only capture and kill all kinds of microorganisms, playing the role of host defense, but also participate in the occurrence and development of diseases.Moreover, EET is also found in bronchial asthma.This paper reviews the structure and formation mechanism of EET and the relationship with asthma.

目的:探讨小鼠间充质干细胞来源的外泌体(exosome,Exo)对D-氨基半乳糖(D-galactosamine hydrochlo-ride,D-GalN)/脂多糖(lipopolysaccharides,LPS)诱导的急性肝损伤治疗作用及可能机制.方法:提取小鼠间充质干细胞,通过流式细胞术和细胞分化实验进行验证.超速离心法提取间充质干细胞外泌体,透射电镜和蛋白质免疫印迹对其鉴定,纳米颗粒追踪分析测定外泌体粒径电位.通过活性氧荧光探针考察外泌体减轻活性氧损伤的功能.CCK-8检测外泌体对肝细胞的保护作用.腹腔注射D-GalN/LPS制备急性肝损伤小鼠模型,考察外泌体对D-GalN/LPS诱导的急性肝损伤小鼠的治疗作用.结果:原代间充质干细胞表达CD29和CD44两个标志蛋白,而不表达CD31和CD117,具有成骨分化能力.间充质干细胞外泌体为具有典型杯状结构的纳米囊泡,表达CD9和CD63两个标志性蛋白.与PBS组相比,干细胞外泌体能够减轻L02细胞内的活性氧水平,提高L02细胞活性,且外泌体本身对L02细胞活性没有影响.与PBS组相比,Exo治疗组能够改善肝脏组织损伤,降低转氨酶水平,调节丙二醛、过氧化氢酶和超氧化物歧化酶的水平.结论:在细胞实验中,间充质干细胞外泌体能够减少活性氧,保护肝细胞的活性;在动物实验中,间充质干细胞外泌体能够下调氧化应激相关蛋白的表达,促进肝脏组织修复.

Objective To investigate the effect of saikosaponin D (SSD) on the proliferation and transformation of human embryonic lung fibroblasts (HELFs) induced by transforming growth factor-beta 1 (TGF-β1) and the regulation of signal pathway of TGF-β1/Smads family. Methods HELFs were cultured in vitro and divided into 5 groups: a control group, 1 ng/mL TGF-β1-induced group, 1 ng/mL TGF-β1 combined with 0.5 μmol/L SSD treatment group, 1 ng/mL TGF-β1 combined with 1 μmol/L SSD treatment group, and 1 ng/mL TGF-β1 combined with 2 μmol/L SSD treatment group. Cell viability of HELFs was detected by CCK-8 assay. The expression of Smad2, Smad3 and Smad7 mRNA were detected by real-time fluorescence quantitative PCR. The protein levels of α-smooth muscle actin (α-SMA), type 1 collagen (Col1), Smad2, Smad3, phosphorylated Smad2 (p-smad2), p-smad3 and Smad7 were assessed by Western blot analysis. Results Compared with the control group, TGF-β1-induced group showed the apparently increased proliferation ability, the increased protein levels of Col1 and α-SMA, the significantly increased mRNA and protein phosphorylation levels of Smad2 and Smad3, and the significantly decreased mRNA and protein expression of Smad7. Compared with the TGF-β1-induced group, the cell proliferation of HELFs in different concentrations of SSD treatment groups was reduced, which could reverse the changes of the above indicators in a dose-dependent manner. Conclusion SSD plays an important role in anti-pulmonary fibrosis by regulating TGF-β1/Smads signaling pathway.

目的:探讨蛋白激酶CK2(casein kinase 2,曾称酪蛋白激酶II)抑制剂CX4945对顺铂(cisplatin,DDP)耐药性肺癌A549/DDP细胞生长活性的影响及其可能的作用机制.方法:CCK-8法检测肺癌A549细胞及其耐药性A549/DDP细胞的DDP半数抑制浓度(half maximal inhibitory concentration,IC50),以及抑制剂CX4945对A549/DDP细胞耐药性的影响.蛋白质印迹法检测A549及A549/DDP细胞在DDP干预前后Wnt信号通路相关蛋白(CK2α、β-catenin和cyclin D1)、耐药相关蛋白[多药耐药相关蛋白1(multidrug resistance-associated protein 1,MRP1)和肺耐药相关蛋白(lung resistance-related protein,LRP)]和凋亡相关蛋白[剪切型半胱天冬酶-3(cleaved caspase-3,c-caspase-3)]的表达变化.将A549/DDP细胞分为未处理对照组、CX4945组、DDP组及CX4945+DDP组,各组药物处理后,采用蛋白质印迹法和F℃M法分别检测Wnt信号通路、耐药及凋亡相关蛋白的表达以及细胞凋亡情况.结果:DDP对A549/DDP细胞的IC50值是A549细胞的4.59倍,并在CX4945预处理后A549/DDP细胞耐药性明显降低(P＜0.001).与A549细胞相比,A549/DDP细胞中CK2α、β-catenin、cyclin D1、MRP1和LRP蛋白的表达水平明显较高,并在DDP处理后进一步升高(P值均＜0.001);然而A549细胞中,DDP处理后CK2α、β-catenin和cyclin D1蛋白表达水平明显降低(P值均＜0.01),而MRP1和LRP蛋白表达则无明显变化(P值均＞0.05).与未处理对照组和DDP组相比,CX4945组和CX4945+DDP组的A549/DDP细胞中β-catenin、cyclin D1、MRP1和LRP蛋白表达水平均明显降低(P值均＜0.01).此外,CX4945+DDP组的A549/DDP细胞凋亡率和c-caspase-3表达水平均较未处理对照组和DDP组明显升高(P值均＜0.001).结论:CK2抑制剂CX4945可通过抑制Wnt通路,降低耐药相关蛋白的表达,从而逆转肺癌A549/DDP细胞的顺铂耐药性.

目的:探讨柴胡皂甙D(SSd)对人肺腺癌A549细胞上皮间质转化(epithelial mesenchymal transition,EMT)的影响及可能机制.方法:体外培养A549细胞,经TGF-β1(10 ng/mL)刺激诱导EMT,观察不同浓度SSd(0.5,1.0,2.0μmol/L)的抑制作用.采用MTT法检测细胞增殖活性,蛋白质印迹检测A549细胞mTOR、p70S6、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、N-钙黏蛋白、Ⅰ型胶原蛋白、E-钙黏蛋白的表达水平.结果:MTT检测结果显示,TGF-β1刺激后A549细胞增殖能力增强,SSd可明显抑制TGF-β1刺激的细胞增殖能力,且呈剂量依赖关系.经SSd作用后,TGF-β1刺激的A549细胞mTOR及下游的p70S6蛋白表达明显下调,α-SMA、Ⅰ型胶原蛋白、N-钙黏蛋白的表达水平明显降低,而E-钙黏蛋白则明显升高,且呈明显的剂量—时间依赖关系.结论:SSd可通过mTOR信号通路抑制A549细胞EMT过程.

Objective:To investigate the effect of sodium tanshinoneⅡA sulfonate(STS) on the pro-liferation of rat pulmonary arterial smooth muscle cells(PASMC) induced by hypoxia and its potential mechanism. Methods:Rat PASMC were selected for the following experiments. First,PASMC were di-vided into normoxia control group,hypoxia group(3% O2),normoxia control +STS low-, medium- and high-dose groups(5,10,20 ng/mL). After cultured for 60 h, the proliferation of PASMC was deter-mined by CCK-8. Then,PASMC were divided into normoxia control group,hypoxia group(3% O2),hy-poxia +STS group(10 ng/mL),qRT-PCR was used to detect the mRNA expressions of mTOR,eIF2α, c-myc. In mechanism study test,PASMC were divided into normoxia control group, hypoxia group(3% O2),hypoxia +STS group(10 ng/mL), hypoxia +rapamycin group(20 nmol/L); Western blotting were used to investigate the expressions of mTOR, p-mTOR, eIF2α, p-eIF2α, c-myc in PASMC. Re-sults:Compared with normoxia control group,cell viability in hypoxia group increased significantly(P<0.05); compared with hypoxia group, cell viability in STS group decreased significantly(P <0.05). qRT-PCR showed the mRNA expression of mTOR,eIF2α and c-myc in hypoxia group increased signifi-cantly in contrast to normoxia control group(P<0.05);compared with hypoxia group,the three mRNAs expression in STS group decreased significantly(P<0.05). In Western blotting,compared with normox-ia control group,the expression of mTOR, p-mTOR, eIF2α, p-eIF2α, c-myc increased significantly in hypoxia group(P<0.05); while in contrast to hypoxia group, the expression of mTOR, p-mTOR, eIF2α, p-eIF2α,c-myc decreased significantly(P<0.05);there was no statistical significance between hypoxia+STS group and hypoxia+rapamycin group(P>0.05). Conclusion:STS could inhibit hypoxi-a-induced proliferation of PASMC by suppressing mTOR/eIF2α signaling pathway.

IL-33 played an important role in inflammatory diseases as evidenced by their high levels of expression in diseased tissues. Previous studies showed that IL-33/ST2L signal transduction pathway participated in epithelial-mesenchymal transition (EMT) of A549 cells. Cytokine IL-1β can increase the expression of MMPs by activating NF-kB. The excessive or inappropriate expression of MMP-9 may randomly and non-selectively destroy the extracellular matrix. TIMP-1 (tissue inhibitor of MMP-9) effects on ebb and flow of ECM by inhibiting activation of MMP-9. Therefore, IL-33 may take part in the process of pulmonary fibrosis by regulating expressions of MMP-9 and TIMP-1. To explore the acting mechanism of IL-33 in pulmonary fibrosis, proliferation of the human embryonic lung fibroblasts and expressions of related signal molecules was analyzed in vitro. We cultured HELF cells and stimulated HELF with rhIL-33 at different time points (24, 48, 72 h) and different concentrations respectively. The expression of the receptor ST2L was analyzed by RT-PCR and the proliferative rate of HELF was tested by MTT. The expressions of collagen IV, MMP-9, TIMP-1, and critical signal transducer TRAF-6 and NF-kappaB were tested by Western blotting. The rhIL-33 can promote proliferation of HELF and the concentration of 10 ng/ml was most significant at 72 h (P < 0.05). Hence, this experiment chose 10 ng/ml as stimulated concentration at following experiments. The expressions of collagen IV, MMP-9, TIMP-1, TRAF-6, and NF-kappaB increased and then reduced in protein levels at different time points (0, 6, 12, 24, 48, 72 h) (P < 0.05). IL-33 participates in the production of profibrotic cytokines and formation of mesenchymal substances in early inflammatory responses of pulmonary fibrosis. IL-33 can regulate deposition of ECM and promote the process of pulmonary fibrosis by inducing the imbalance between MMP-9 and TIMP-1.

Schisandrin B (Sch B), a dibenzocyclooctadiene lignan isolated from Schisandra chinensis (Turcz.) Baill , has been shown to have anti-inflammatory effect. The purpose of this study was to evaluate the effect of Sch B on LPS-induced inflammation in microglia and to investigate the molecular targets of Sch B. BV2 cells were stimulated by LPS in the presence or absence of Sch B. The results showed that the levels of TNF-α, IL-6, IL-1β, and PGE 2 upregulated by LPS were significantly suppressed by Sch B. LPS-induced NF-κB activation was also inhibited by Sch B. Furthermore, Sch B was found to upregulate the expression of PPAR-γ in a concentration-dependent manner. In addition, the inhibition of Sch B on TNF-α, IL-6, IL-1β, and PGE 2 production were reversed by PPAR-γ antagonist GW9662. In conclusion, these results suggested that Sch B inhibited LPS-induced inflammatory response by activating PPAR-γ.

Objective:By constructing β-catenin siRNA lentiviral vectors, the correlation of Wnt pathway with cisplatin resistance of non-small cell lung cancer was explored.Methods:According to the β-catenin gene mRNA sequence, we designed and synthesized three pairs for β-catenin gene shRNA1, shRNA2, shRNA3.Forming double stranded DNA and pLentiLox3.7 lentiviral vector were connected and transformed.The positive clones were selected and the shRNA constructor was identified by DNA sequencing.The recombinant lentivirus was obtained by packaging cells 293T.The titer of virus was tested according to the expression level of GFP.A549 cells were infected to screen out the effective inhibition of the target.After cisplatin-resistant cell line A549/DDP were infected by lentivirus in 24 h, different concentrations of cisplatin were used to interfere with the cells 48 h, MTT assay was used to detect the cells proliferation.According to the cells were infected by lentivirus or not, divided into A549 group, A549/DDP group, A549/DDP+ siRNA group, A549/DDP+siNC group.After 8μg/mL cisplatin was used to interfere with the cells in each group, the expression of LC3 Ⅱ and β-catenin was tested by real-time PCR and western blotting.Results:DNA sequencing showed that the β-catenin shRNA lentiviral vector was constructed successfully and screen out the effective inhibition of the target.At the same concentration, the inhibitory effect of cisplatin on A549 cells was significantly higher than that of A549/DDP cells(P<0.05).After siRNA lentivirus infecting A549/DDP cells, the inhibitory effect of cisplatin on A549/DDP cells was increased.Compared with A549 group, treated with 8 μg/mL cisplatin for 48 h, the expression levels of β-catenin mRNA and protein in A549/DDP group were increased, meanwhile the expression of LC3Ⅱ mRNA and protein were also increased(P<0.05).Compared with A549/DDP group, after treating with cisplatin, the expression levels of β-catenin mRNA and protein in A549/DDP and siRNA cells group were decreased, and the expression of LC3 Ⅱ mRNA and protein were also decreased.Conclusion:The Wnt pathway played an important role in the formation of the cisplatin resistence in non-small cell lung cancer, which was associated with autophagy.

Purpose: To investigate the therapeutic effects of saikosapoin D (SSD) on bleomycin (BLM)-induced pulmonary fibrosis (PF) in mice and its probable mechanisms.Methods: PF mice were prepared by intraperitoneal (i.p.) injection of BLM (5 mg/kg). Twenty-four hours later, 72 mice in SSD group were administered SSD (1.8 mg/kg, ip). After 3, 7, 14 and 28 days of injection, the mice were sacrificed. Blood samples and lung tissues were collected from 6 mice in each group. The lung tissues were subjected to histological examination. In addition, expressions of MyD88, TRAF6, IL-33 and ST2 in lung tissue were determined by western blotting assay. Serum levels of hydroxyproline (HYP), interleukin (IL)-4, IL-13 and interferon (IFN)-y were measured by enzyme-linked immunosorbent assay (ELISA).Results: Pathological results showed that SSD treatment alleviated alveolitis and lung fibrosis (p < 0.05) in lung tissues of PF mice at 14 and 28 days post-BLM injection. HYP and IL-13 levels of mice in SSD group were significantly lower than that in BLM group at days 14 and 28 post-BLM injection (p < 0.05). Levels of IL-4 and IFN-y were significantly lower when compared with values in BLM group on day 28 (p < 0.05). Western blotting results revealed that expressions of MyD88, TRAF6, IL-33 and ST2 proteins were significantly decreased by SSD treatment (p < 0.05).Conclusion: SSD exerts therapeutic effects on BLM-induced experimental PF in mice via regulation of IL-33/ST2 pathway.