Intratracheal administration of umbilical cord-derived mesenchymal stem cells attenuates hyperoxia-induced multi-organ injury via heme oxygenase-1 and JAK/STAT pathways

BACKGROUND Bronchopulmonary dysplasia(BPD)is not merely a chronic lung disease,but a systemic condition with multiple organs implications predominantly associated with hyperoxia exposure.Despite advances in current management strategies,limited progress has been made in reducing the BPD-related systemic damage.Meanwhile,although the protective effects of human umbilical cord-derived mesenchymal stem cells(hUC-MSCs)or their exosomes on hyperoxia-induced lung injury have been explored by many researchers,the underlying mechanism has not been addressed in detail,and few studies have focused on the therapeutic effect on systemic multiple organ injury.AIM To investigate whether hUC-MSC intratracheal administration could attenuate hyperoxia-induced lung,heart,and kidney injuries and the underlying regulatory mechanisms.METHODS Neonatal rats were exposed to hyperoxia(80%O_(2)),treated with hUC-MSCs intratracheal(iT)or intraperitoneal(iP)on postnatal day 7,and harvested on postnatal day 21.The tissue sections of the lung,heart,and kidney were analyzed morphometrically.Protein contents of the bronchoalveolar lavage fluid(BALF),myeloper oxidase(MPO)expression,and malondialdehyde(MDA)levels were examined.Pulmonary inflammatory cytokines were measured via enzyme-linked immunosorbent assay.A comparative transcriptomic analysis of differentially expressed genes(DEGs)in lung tissue was conducted via RNA-sequencing.Subsequently,we performed reverse transcription-quantitative polymerase chain reaction and western blot analysis to explore the expression of target mRNA and proteins related to inflammatory and oxidative responses.RESULTS iT hUC-MSCs administration improved pulmonary alveolarization and angiogenesis(P<0.01,P<0.01,P<0.001,and P<0.05 for mean linear intercept,septal counts,vascular medial thickness index,and microvessel density respectively).Meanwhile,treatment with hUC-MSCs iT ameliorated right ventricular hypertrophy(for Fulton’s index,P<0.01),and relieved reduced nephrogenic zone width(P<0.01)and glomerular diameter(P<0.001)in kidneys.Among the beneficial effects,a reduction of BALF protein,MPO,and MDA was observed in hUC-MSCs groups(P<0.01,P<0.001,and P<0.05 respectively).Increased pro-inflammatory cytokines tumor necrosis factor-alpha,interleukin(IL)-1β,and IL-6 expression observed in the hyperoxia group were significantly attenuated by hUC-MSCs administration(P<0.01,P<0.001,and P<0.05 respectively).In addition,we observed an increase in anti-inflammatory cytokine IL-10 expression in rats that received hUC-MSCs iT compared with rats reared in hyperoxia(P<0.05).Transcriptomic analysis showed that the DEGs in lung tissues induced by hyperoxia were enriched in pathways related to inflammatory responses,epithelial cell proliferation,and vasculature development.hUC-MSCs administration blunted these hyperoxia-induced dysregulated genes and resulted in a shift in the gene expression pattern toward the normoxia group.hUC-MSCs increased heme oxygenase-1(HO-1),JAK2,and STAT3 expression,and their phosphorylation in the lung,heart,and kidney(P<0.05).Remarkably,no significant difference was observed between the iT and iP administration.CONCLUSION iT hUC-MSCs administration ameliorates hyperoxia-induced lung,heart,and kidney injuries by activating HO-1 expression and JAK/STAT signaling.The therapeutic benefits of local iT and iP administration are equivalent.

Leptin activates the JAK/STAT pathway to promote angiogenesis in RF/6A cells in vitro

AIM: To investigate the effect of leptin on the angiogenesis of RF/6 A cells(monkey retinal choroidal endothelial cells) in vitro and test the cellular signaling in the mechanism.METHODS: RF/6 A cells were cultured in vitro and randomly divided into four groups: normal control, with leptin at 50, 100, 200 ng/m L for cell counting kit-8(CCK8). RF/6 A cell proliferation and migration were examined by Transwell assays, while RF/6 A cell tube formation by Matrigel assay. JAK2, p-JAK2, STAT3, and p-STAT3 protein expression was measured by Western blotting. Cells were then divided into the following treatment groups: control, 100 ng/m L leptin and AG-490(100 ng/m L leptin+10 μmol/L AG-490) for examinations of RF/6 A cellular behaviour again. Analysis of differences was carried out using one-way ANOVA and least significant difference(LSD).RESULTS: RF/6 A cell proliferation, migration and cell tube formation were promoted significantly by leptin in a dose-dependent manner(P<0.05). Western blotting showed that leptin up-regulated p-JAK2 and p-STAT3 expression levels. Treatment with the JAK/STAT pathway inhibitor, AG-490, decreased leptin-induced p-JAK2 and p-STAT3 expression, and inhibited cell proliferation, migration and cell tube formation induced by leptin(P<0.05). CONCLUSION: Leptin can promote RF/6 A cell angiogenesis in vitro via activation of the JAK2/STAT3 signaling pathway.

Argatroban promotes recovery of spinal cord injury by inhibiting the PAR1/JAK2/STAT3 signaling pathway

Argatroban is a synthetic thrombin inhibitor approved by U.S.Food and Drug Administration for the treatment of thrombosis.However,whether it plays a role in the repair of spinal cord injury is unknown.In this study,we established a rat model of T10 moderate spinal cord injury using an NYU Impactor ModerⅢand performed intraperitoneal injection of argatroban for 3 consecutive days.Our results showed that argatroban effectively promoted neurological function recovery after spinal cord injury and decreased thrombin expression and activity in the local injured spinal cord.RNA sequencing transcriptomic analysis revealed that the differentially expressed genes in the argatroban-treated group were enriched in the JAK2/STAT3 pathway,which is involved in astrogliosis and glial scar formation.Western blotting and immunofluorescence results showed that argatroban downregulated the expression of the thrombin receptor PAR1 in the injured spinal cord and the JAK2/STAT3 signal pathway.Argatroban also inhibited the activation and proliferation of astrocytes and reduced glial scar formation in the spinal cord.Taken together,these findings suggest that argatroban may inhibit astrogliosis by inhibiting the thrombin-mediated PAR1/JAK2/STAT3 signal pathway,thereby promoting the recovery of neurological function after spinal cord injury.

RNA结合蛋白KHSRP激活JAK1/STAT3通路促进胃贲门腺癌的生长与转移

目的探讨KHSRP靶向调控JAK1/STAT3信号轴对胃贲门腺癌细胞迁移和侵袭等恶性生物学行为的影响。方法比较胃贲门腺癌组织和癌旁组织中KHSRP的表达水平。qRT-PCR法检测胃贲门腺癌细胞系(OE-19、TE-7、BIC-1、FLO-1、SK-GT-4、BE-3)与正常胃黏膜上皮细胞系(GES-1)中KHSRP的表达差异。通过包装慢病毒载体对KHSRP进行敲降和过表达处理,分为sh-NC组、sh-KHSRP组和Vector组、KHSRP组。采用细胞计数试剂盒-8(CCK-8)、Transwell实验测定KHSRP对胃贲门腺癌细胞增殖、迁移和侵袭的影响。异种移植肿瘤模型检测敲降和过表达KHSRP在活体动物体内的作用。Western blot实验验证KHSRP靶向JAK/STAT信号通路。结果与邻近正常组织相比,KHSRP在GCA组织中的表达显著增高(P<0.05)。细胞功能实验分析显示,KHSRP过表达在体外显著促进GCA细胞增殖、迁移和侵袭(P<0.05)。在敲降KHSRP后,JAK/STAT信号通路中JAK1、STAT3磷酸化水平明显降低,过表达KHSRP后情况则反之(P<0.05)。结论KHSRP通过激活JAK1/STAT3信号轴调控胃贲门腺癌转移的恶性进程。

Metochalcone induces senescence-associated secretory phenotype via JAK2/STAT3 pathway in breast cancer

Breast and lung cancers are the leading causes of mortality and most frequently diagnosed cancers in women and men,respectively,worldwide.Although the antitumor activity of chalcones has been extensively studied,the molecular mechanisms of isoliquiritigenin analog 2',4',4-trihydroxychalcone(metochalcone;TEC)against carcinomas remain less well understood.In this study,we found that TEC inhibited cell proliferation of breast cancer BT549 cells and lung cancer A549 cells in a concentration-dependent manner.TEC induced cell cycle arrest in the S-phase,cell migration inhibition in vitro,and reduced tumor growth in vivo.Moreover,transcriptomic analysis revealed that TEC modulated the activity of the JAK2/STAT3 and P53 pathways.TEC triggered the senescence-associated secretory phenotype(SASP)by repressing the JAK2/STAT3 axis.The mechanism of metochalcone against breast cancer depended on the induction of SASP via deactivation of the JAK2/STAT3 pathway,highlighting the potential of chalcone in senescence-inducing therapy against carcinomas.

JAK/STAT信号通路与肌肉骨骼退行性疾病的研究进展

JAK抑制剂具有强大的抗炎作用,目前有多项研究发现了JAK抑制剂对MSDDs的实验模型具有治疗作用。本文将从JAK/STAT信号通路、JAK抑制剂与MSDDs之间的研究进展进行综述,旨在为此类疾病提供新的治疗思路。

Yangyin Huowei mixture alleviates chronic atrophic gastritis by inhibiting the IL-10/JAK1/STAT3 pathway

BACKGROUND The Chinese medicine Yangyin Huowei mixture(YYHWM)exhibits good clinical efficacy in the treatment of chronic atrophic gastritis(CAG),but the mechanisms underlying its activity remain unclear.AIM To investigate the therapeutic effects of YYHWM and its underlying mechanisms in a CAG rat model.METHODS Sprague-Dawley rats were allocated into control,model,vitacoenzyme,and low,medium,and high-dose YYHWM groups.CAG was induced in rats using Nmethyl-N′-nitro-N-nitrosoguanidine,ranitidine hydrochloride,hunger and satiety perturbation,and ethanol gavage.Following an 8-wk intervention period,stomach samples were taken,stained,and examined for histopathological changes.ELISA was utilized to quantify serum levels of PG-I,PG-II,G-17,IL-1β,IL-6,and TNF-α.Western blot analysis was performed to evaluate protein expression of IL-10,JAK1,and STAT3.RESULTS The model group showed gastric mucosal layer disruption and inflammatory cell infiltration.Compared with the blank control group,serum levels of PGI,PGII,and G-17 in the model group were significantly reduced(82.41±3.53 vs 38.52±1.71,23.06±0.96 vs 11.06±0.70,and 493.09±12.17 vs 225.52±17.44,P<0.01 for all),whereas those of IL-1β,IL-6,and TNF-αwere significantly increased(30.15±3.07 vs 80.98±4.47,69.05±12.72 vs 110.85±6.68,and 209.24±11.62 vs 313.37±36.77,P<0.01 for all),and the protein levels of IL-10,JAK1,and STAT3 were higher in gastric mucosal tissues(0.47±0.10 vs 1.11±0.09,0.49±0.05 vs 0.99±0.07,and 0.24±0.05 vs 1.04±0.14,P<0.01 for all).Compared with the model group,high-dose YYHWM treatment significantly improved the gastric mucosal tissue damage,increased the levels of PGI,PGII,and G-17(38.52±1.71 vs 50.41±3.53,11.06±0.70 vs 15.33±1.24,and 225.52±17.44 vs 329.22±29.11,P<0.01 for all),decreased the levels of IL-1β,IL-6,and TNF-α(80.98±4.47 vs 61.56±4.02,110.85±6.68 vs 89.20±8.48,and 313.37±36.77 vs 267.30±9.31,P<0.01 for all),and evidently decreased the protein levels of IL-10 and STAT3 in gastric mucosal tissues(1.11±0.09 vs 0.19±0.07 and 1.04±0.14 vs 0.55±0.09,P<0.01 for both).CONCLUSION YYHWM reduces the release of inflammatory factors by inhibiting the IL-10/JAK1/STAT3 pathway,alleviating gastric mucosal damage,and enhancing gastric secretory function,thereby ameliorating CAG development and cancer transformation.

Oleanolic acid inhibits colon cancer cell stemness and reverses chemoresistance by suppressing JAK2/STAT3 signaling pathway

Background:Oleanolic acid(OA),a pentacyclic triterpenoid exhibiting specific anti-cancer properties and highly effective antioxidant activity,was isolated from traditional Chinese medicinal herbs.Conversely,the OA that impacts colon cancer(CC)cells and its underlying mechanisms remain poorly understood.Methods:The cytotoxic effect of OA alone or OA-5-Fluorouracil(5-FU)combination on normal and CC cells was analyzed by methyl thiazolyl diphenyl-tetrazolium bromide(MTT).Then,the impact of OA on CC cell lines(LoVo and HT-29)proliferation and stemness were measured using colon formation and tumorsphere formation assays.Octamer-binding transcription factor 4(Oct4),Prominin-1(CD133),Nanog,and transcription factor SOX-2(SOX2)are cell stemness-related indicators whose expression was assessed usingfluorescence qPCR assay,Western blotting,and immunohistochemistry.The effect of OA on the proliferative potency of CC cells was evaluated using an in vivo model.Results:The stem-like characteristics and clone production of colon cancer cells were markedly reduced by OA alone or in combination with OA-5-FU.Moreover,OA increases the susceptibility of CC cells to 5-FU by blocking the cell stemness-related markers(CD133,Nanog,SOX2,and Oct4)expression levels both in vitro and in vivo,as well as by inactivating the activator of transcription 3(STAT3 signaling)and Janus kinase 2/signal transducer(JAK2).Conclusion:Thesefindings imply that oleanolic acid,both in vitro and in vivo,suppresses the JAK2/STAT3 pathway,which in turn reverses chemoresistance and decreases colon cancer cell stemness.Therefore,by reducing the recommended amount of 5-FU,this strategy may improve chemotherapeutic effectiveness and minimize undesired side effects.

CDK8 Promotes Cell Proliferation, Migration and Invasion in Esophageal Squamous Cell Carcinoma Through JAK/ STAT3/EMT Pathway

Objective To investigate the expression of cyclin-dependent kinase 8(CDK8)in esophageal squamous cell carcinoma(ESCC)and its effect on ESCC cells,and to explore its potential molecular mechanism.Methods The expression level of CDK8 mRNA was analyzed using UALCAN database,and then the expression level of CDK8 protein in tumor tissues of ESCC patients was detected by immunohistochemistry(IHC).Esophageal cancer cell lines Kyse-30 and Kyse-150 were stably transfected with lentivirus to achieve knockdown and overexpression of CDK8.EdU proliferation assay,cell colony formation assay,cell cycle assay,cell scratch assay and invasion assay were used to explore the effect of CDK8 protein expression level on the phenotype of ESCC cells.Subsequently,the effect of CDK8 on the growth of esophageal cancer xenografts in vitro was observed by subcutaneous tumor formation assay in mice.Finally,the expression of proliferation and metastasis related proteins was detected by Western blot.Results CDK8 showed high transcription and protein expression levels in ESCC tissues compared with normal esophageal tissues.Knockdown of CDK8 expression significantly inhibited the proliferation,migration and invasion of ESCC cells.In addition,inhibition of CDK8 expression significantly affected the JAK2/STAT3 pathway and the expression of E-cadherin/N-cadherin,while overexpression of CDK8 reversed these effects.Inhibition of STAT3 pathway reversed the promoting effect of CDK8 overexpression on ESCC cell phenotype.Conclusion CDK8 is a cancer-promoting factor of ESCC,which mediates the phosphorylation of JAK2/STAT3 and epithelial-mesenchymal transition(EMT).

Exploring the mechanism of electroacupuncture at different acupoints on acute colitis rats based on JAK2/STAT3/SOCS1 signaling pathway

Objective:To investigate the mechanism of JAK2/STAT3/SOCS1 signaling pathway in electroacupuncture of different acupoints on acute colitis rats.Methods:36 SPF SD rats were randomly divided into 6 groups,with 6 rats in each group.The rat model of acute colitis was prepared by enema with glacial acetic acid solution.After the model was established,electroacupuncture was given to each acupoint group,with density wave,frequency 2Hz-50 Hz,intensity 2 mA,muscle tremor as the degree 20 min/time,1 time/day,for 3 consecutive days.Observe the general condition of rats;the pathological changes of colonic mucosa in rats were observed by HE method.The contents of serum interleukin-4(IL-4)and interleukin-8(IL-8)were detected by ELISA.Western blot and RT-PCR were used to detect the expression of JAK2,STAT3,SOCS1 protein and mRNA in rat colon tissue.Results:In contrast to the normal group,the overall condition of the model group was worse,the colonic mucosa was severely damaged,even necrotic,and the ulcer surface was obvious.The content of IL-4 in serum was obviously reduced,and the content of IL-8 was obviously go up(P<0.01).The protein content of JAK2,STAT3 and the expression of JAK2,STAT3 mRNA in colon tissue of rats were obviously go up,while the protein content of SOCS1 and the expression of SOCS1 mRNA were obviously reduced(P<0.01).In contrast to the model group,the general condition of rats in each acupoint group was significantly improved,the damage and necrosis of colonic mucosa and ulcer surface were obviously alleviated,the content of IL-4 in serum was obviously go up,and the content of IL-8 was significantly decreased(P<0.01).The protein content of JAK2,STAT3 and the expression of JAK2,STAT3 mRNA in colon tissue of rats were obviously reduced,while the protein content of SOCS1 and the expression of SOCS1 mRNA were obviously go up(P<0.05,P<0.01).Comparison of different acupoint groups,the colonic mucosal injury in the Zusanli group was significantly reduced,the content of serum IL-4 was significantly increased,and the content of IL-8 was significantly decreased(P<0.05,P<0.01).The protein content and mRNA expression of JAK2 and STAT3 in colon tissue were significantly down-regulated,while the protein content and mRNA expression of SOCS1 were significantly go up(P<0.05,P<0.01).Conclusion:Electroacupuncture at each acupoint can improve the damage of colonic mucosa and reduce the inflammatory response.The therapeutic effect of Zusanli(ST36)is better than that of Tianshu(ST25),Dachangshu(BL25)and Shangjuxu(ST37).The mechanism may be related to the regulation of JAK2/STAT3/SOCS1 signaling pathway related proteins and inflammatory cytokines IL-4 and IL-8.

基于JAK/STAT信号通路探讨针刺治疗脑出血机制研究进展

脑出血是一种急性脑血管病,发病率、病死率较高,发病机制十分复杂。Janus酪氨酸蛋白激酶(JAK)/信号转导及转录激活因子(STAT)信号通路在脑出血发展过程中起关键作用。针刺治疗脑出血疗效肯定,本文以JAK/STAT信号通路作为切入点,梳理近年来针刺治疗脑出血机制研究,归纳其在抑制炎性反应,减轻脑水肿,抑制细胞凋亡,促进神经、血管再生、促进神经功能重塑等方面作用,为相关研究提供参考。

真武汤加减治疗中重度变应性鼻炎的疗效及对JAK2/STAT信号通路的影响

目的:观察真武汤加减治疗中重度变应性鼻炎肾阳虚证的疗效及对JAK2/STAT信号通路的影响。方法:120例变应性鼻炎患者随机分为2组,每组60例。对照组给予糠酸莫米松联合盐酸左西替利嗪,观察组口服真武汤加减,治疗4周。比较两组患者鼻症状积分(TNSS)、圣乔治呼吸问卷(SGRQ)、中医症状、鼻腔最小横截面积(NMCA)、0~7 cm阶段鼻腔容积(NCV_(0-7))、吸气过程中鼻通气阻力(iNAR)和呼气过程中鼻通气阻力(eNAR)、血清免疫功能指标(IgE、IgG、IgA、IgM)、血清和鼻分泌液中炎性因子(EOT、CCR3、EOS、IL-17)及鼻分泌液中JAK2、STAT3、STAT4、STAT5水平。比较两组临床疗效及不良反应。结果:观察组总有效率96.3%,显著高于对照组的80.4%(P<0.05)。治疗后,与对照组比较,观察组TNSS、SGRQ、中医症状评分及iNAR、eNAR水平显著降低(P<0.05),NMCA、NCV_(0-7)、IgG、IgA、IgM水平显著升高,IgE、EOT、CCR3、EOS、IL-17、JAK2、STAT3、STAT4、STAT5水平显著降低(P<0.05)。观察组不良反应发生率(1.7%)显著低于对照组(23.2%)(P<0.05)。结论:真武汤加减可明显缓解中重度变应性鼻炎肾阳虚证患者的鼻部症状,其作用机制可能与调节JAK2/STAT信号通路有关。

香青兰总黄酮调控TMAO介导的JAK/STAT轴抗大鼠动脉粥样硬化及RAW264.7细胞炎症的研究

目的探讨香青兰总黄酮(total flavonoids of Dracocephalum Moldavica L.,TFDM)抗大鼠动脉粥样硬化(atherosclerosis,AS)及三甲胺氮氧化物(trimethylamine N-oxide,TMAO)加剧的小鼠巨噬细胞RAW264.7炎症的保护作用及其可能的作用机制。方法采用高脂饲料喂养联合腹腔注射维生素D3的方法建立SD大鼠AS模型,分为对照组、模型组、辛伐他汀组(15 mg·kg^(-1))、TFDM组(60、30、15 mg·kg^(-1)),建模同时进行给药处理,持续8周。生化检测方法检测血清总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)水平。HE染色检测主动脉组织病理变化,ELISA试剂盒检测血清中TMAO、IL-1β、IL-6及肝脏组织中TNF-α的表达,Western blot法检测主动脉中JAK、STAT、TNF-α蛋白的表达。此外,体外培养RAW264.7巨噬细胞,采用LPS+TMAO建立巨噬细胞炎症模型,TFDM(100、50、25 mg·L^(-1))进行干预。CCK-8测定细胞活力及增殖,RT-qPCR法检测细胞中TNF-α、IL-6、JAK、STAT mRNA的表达。结果TFDM可明显下调大鼠血清TC、TG、LDL-C水平及血清TMAO、IL-1β、IL-6和肝脏TNF-α的水平,减少主动脉的斑块沉积,下调主动脉中TNF-α、JAK、STAT的蛋白表达。此外,TFDM干预可明显下调TNF-α、IL-6、JAK、STAT mRNA的表达及JAK、STAT蛋白的表达。结论TFDM可降低血清中TMAO的含量,从而抑制JAK/STAT炎症信号通路,减缓炎症的发生,发挥抗AS作用。

类风湿关节炎的JAK/STAT信号通路文献计量分析研究

目的通过对近十年Janus激酶(Janus kinase,JAK)/信号转导和转录激活因子(signal transducer and activator of transcription,STAT)信号通路对类风湿关节炎作用的文献进行多软件可视化分析,总结该领域发展趋势和研究热点,为研究者提供新的方向和思路,促进该领域创新性发展。方法收集Web of Science Core Collection数据库2013至2023年JAK/STAT信号通路在类风湿关节炎中的相关文献。利用CiteSpace和VOSviewer软件对检索到的354篇文章的发文量、国家、作者、关键词进行分析。结果该领域发文量持续增加,根据作者研究方向、高频词的呈现及对前言和热点的关注,提示该领域聚焦于基因表达、免疫机制、炎症机制、途径抑制剂、药物治疗等。未来研究将围绕通路抑制剂、抗风湿药物的安全性、机制、对照试验等展开。结论JAK/STAT信号通路对类风湿关节炎影响的研究在过去、现在乃至未来关注度高,研究价值大。各团队对该领域的研究存在差异,区域发展不平衡,提示应加强合作与交流、聚焦国际前沿、开展更多高质量研究,以推动该领域的发展和进步,为临床提供依据。

WNT5A drives interleukin-6-dependent epithelial-mesenchymal transition via the JAK/STAT pathway in keloid pathogenesis

Background:Keloid scarring is a fibroproliferative disease caused by aberrant genetic activation with an unclear underlying mechanism.Genetic predisposition,aberrant cellular responses to environmental factors,increased inflammatory cytokines and epithelial-mesenchymal transition(EMT)phenomena are known as major contributors.In this study,we aimed to identify the molecular drivers that initiate keloid pathogenesis.Methods:Bulk tissue RNA sequencing analyses of keloid and normal tissues along with ex vivo and in vitro tests were performed to identify the contributing genes to keloid pathogenesis.An animal model of inflammatory keloid scarring was reproduced by replication of a skin fibrosis model with intradermal bleomycin injection in C57BL/6 mice.Results:Gene set enrichment analysis revealed upregulation of Wnt family member 5A(WNT5A)expression and genes associated with EMT in keloid tissues.Consistently,human keloid tissues and the bleomycin-induced skin fibrosis animal model showed significantly increased expression ofWNT5A and EMT markers.Increased activation of the interleukin(IL)-6/Janus kinase(JAK)/signal transducer and activator of transcription(STAT)pathway and subsequent elevation of EMT markerswas also observed in keratinocytes co-cultured withWNT5A-activated fibroblasts or keloid fibroblasts.Furthermore,WNT5A silencing and the blockage of IL-6 secretion via neutralizing IL-6 antibody reversed hyperactivation of the STAT pathway and EMT markers in keratinocytes.Lastly,STAT3 silencing significantly reduced the EMT-like phenotypes in both keratinocytes and IL-6-stimulated keratinocytes.Conclusions:Intercellular communication via the WNT5A and STAT pathways possibly underlies a partial mechanism of EMT-like phenomena in keloid pathogenesis.IL-6 secreted from WNT5A-activated fibroblasts or keloid fibroblasts activates the JAK/STAT signaling pathway in adjacent keratinocytes which in turn express EMT markers.A better understanding of keloid development and the role of WNT5A in EMT will promote the development of next-generation targeted treatments for keloid scars.

静态磁场通过激活FGFR1/JAK2/STAT3信号通路促进皮肤成纤维细胞的增殖活性和迁移表型

目的:通过体外实验探讨静态磁场(SMF)对皮肤成纤维细胞增殖活性和迁移表型的调控作用与潜在机制。方法:将人皮肤成纤维细胞(HSFs)分为5组,包括对照组、SMF组、SMF+阿魏酸组、SMF+芦可替尼组、SMF+Stattic组。对照组为正常培养的HSFs;SMF组的HSFs细胞暴露于1mT的SMF中。其余三组分别将FGFR1、JAK2、STAT3的抑制剂(3μmoL/L阿魏酸、3nmoL/L芦可替尼、20μmoL/L的Stattic)与HSFs一起预培养,并暴露于1mT的SMF中,所有组均处理24h。用细胞计数试剂盒-8(CCK-8)分析细胞的增殖活性。用ELISA试剂盒法检测人类B细胞淋巴瘤2相关X蛋白(BAX)的水平。用Western blot检测凋亡标志蛋白cleaved-caspase3及FGFR1、JAK2、STAT3、磷酸化的(p)-FGFR1、p-JAK2、p-STAT3的表达以及细胞迁移相关表型高迁移率组蛋白1(HMGB1)、波形蛋白(vimentin)、血管内皮生长因子A(VEGFA)、基质金属蛋白酶-9(MMP-9)、MMP-2的表达。结果:与对照组比,SMF组的细胞增殖活性增加,BAX的水平减少,cleaved-caspase3的蛋白表达水平下调,p-FGFR1、p-JAK2、p-STAT3、HMGB1、vimentin、VEGFA、MMP-9、MMP-2的表达水平均显著上调(均P<0.05)。与SMF组比,SMF+阿魏酸组的细胞增殖活性降低,BAX的水平增加,cleaved-caspase3的蛋白表达水平上调,而p-FGFR1、p-JAK2、p-STAT3、HMGB1、vimentin、VEGFA、MMP-9、MMP-2的表达均显著下调(均P<0.05)。与SMF组比,SMF+芦可替尼组的细胞增殖活性降低,BAX的水平增加,cleaved-caspase3的蛋白表达水平上调,JAK2、p-JAK2、p-STAT3、HMGB1、vimentin、VEGFA、MMP-9、MMP-2的表达水平均显著下调(均P<0.05)。与SMF组比,SMF+Stattic组的细胞增殖活性降低,BAX的水平增加,cleaved-caspase3的蛋白表达水平上调,STAT3、p-STAT3、HMGB1、vimentin、VEGFA、MMP-9、MMP-2的表达水平均显著下调(均P<0.05)。结论:SMF通过激活FGFR1/JAK2/STAT3信号通路促进皮肤成纤维细胞的增殖活性和迁移表型。

基于Hepcidin和JAK2/STAT3信号通路探讨通痹颗粒 对胶原诱导性关节炎大鼠的影响

目的研究通痹颗粒对胶原诱导性关节炎(collagen-induced arthritis,CIA)大鼠铁调素(hepcidin,Hepc)、Janus激酶(janus kinase,JAK)2/信号转导子和转录激活子(signal transduction and activator of transcription,STAT)3信号通路的影响。方法选取36只雌性SD大鼠随机分成空白组、模型组、阳性对照组和通痹颗粒低、中、高剂量组,每组6只。空白组不予处理,其余组用牛Ⅱ型胶原建立CIA模型。造模完成后,空白组、模型组予生理盐水灌胃,其余各组分别以巴瑞替尼片和低、中、高剂量通痹颗粒灌胃。每天1次,连续4周。HE染色行滑膜组织病理学观察;酶联免疫吸附法测定血清Hepc、白细胞介素6(interleukin 6,IL-6)水平;逆转录-聚合酶链反应法测定滑膜中JAK2、STAT3、细胞信号因子传导抑制体(suppressor of cytokine signaling,SOCS)1、SOCS3的mRNA相对表达量;Western blot法检测滑膜中JAK2、p-JAK2、STAT3、p-STAT3、SOCS1、SOCS3的蛋白表达量。结果模型组见滑膜上皮结构缺损,滑膜重度增生,排列紊乱,并有大量炎症细胞浸润和多个血管翳形成;各给药组滑膜炎症均有所减轻,阳性对照组优于通痹颗粒高剂量组,通痹颗粒中、高剂量组优于低剂量组。与模型组相比,各给药组关节炎指数评分、血清Hepc和IL-6水平均显著降低(P<0.01);与阳性对照组相比,通痹颗粒中、低剂量组关节炎指数评分、血清Hepc和IL-6水平均升高(P<0.05)。与模型组比较,阳性对照组和通痹颗粒低、中、高剂量组JAK2、STAT3 mRNA和蛋白以及p-JAK2、p-STAT3的蛋白表达量均降低(P<0.05),而通路抑制因子SOCS1、SOCS3 mRNA和蛋白的表达均升高(P<0.05);与阳性对照组比较,通痹颗粒各剂量组JAK2、STAT3 mRNA和蛋白以及p-JAK2、p-STAT3的蛋白表达量均升高(P<0.05),而SOCS1、SOCS3 mRNA和蛋白的表达均降低(P<0.05)。结论通痹颗粒能够改善CIA大鼠滑膜炎症,其机制可能与抑制JAK2/STAT3信号通路而减少Hepc的表达有关。

纤维粘连蛋白靶向JAK/STAT3信号通路调控粘连性肠梗阻的机制

目的:探究纤维粘连蛋白(FN)靶向JAK/STAT3信号通路调控粘连性肠梗阻的作用机制。方法:选取2019年1月—2020年12月就诊行肠切除手术的粘连性肠梗阻患者65例(观察组),同期行腹股沟疝手术患者65例(对照组),取患者肠组织及术前血液样本,采用qRT-PCR、Western blot及免疫组织化学实验检测肠组织样本中FN、STAT3、p-STAT3基因及蛋白表达,采用酶联免疫吸附实验(ELISA)检测血清样本中FN表达。体外培养人肠上皮细胞系HIEC-6,采用慢病毒转染抑制细胞FN表达,采用CCK8、Transwell迁移实验观察FN对细胞增殖及分化的影响,同时采用qRT-PCR、Western Blot实验检测FN对细胞STAT3、p-STAT3表达的影响。结果:与对照组比较,观察组肠组织中FN mRNA及蛋白表达明显较高(P<0.05),STAT3、p-STAT3 mRNA及蛋白表达则明显较低(P<0.05),且观察组血清中分泌型FN含量也明显较高(P<0.05)。体外实验显示,与NC siRNA组HIEC-6细胞比较,LV-FN siRNA组HIEC-6细胞在48、72、96、120 h的增殖率均明显较低(P<0.05),细胞迁移能力也明显较低(P<0.05),其FN mRNA及蛋白表达明显较低(P<0.05),STAT3、p-STAT3 mRNA及蛋白则明显较高(P<0.05)。结论:纤维粘连蛋白在粘连性肠梗阻患者肠组织及血清中呈明显高表达,可通过JAK/STAT3信号通路的异常激活影响细胞的增殖和迁移,引起病变肠段发病。

藏红花素通过抑制JAK2/STAT3信号通路减轻脑缺血再灌注大鼠海马神经元损伤

目的研究藏红花素(CRO)对脑缺血再灌注(CI/R)大鼠海马神经元损伤的影响并探索其潜在机制。方法将144只雄性SD大鼠按照随机数字表法分为假手术(Sham)组,模型(CI/R)组,CRO低、中、高剂量(CRO-L、CRO-M、CRO-H)组和尼莫地平(NMP)组6组,每组24只。采用线栓法制备CI/R大鼠模型,各组分别于造模前7 d开始1次/d腹腔注射(ip)给药(CRO-L、CRO-M、CRO-H组分别ip给药10、20、40 mg/kg,NMP组ip给药1 mg/kg,Sham组和CI/R组ip给予生理盐水5 mL/kg)。再灌注24 h后,通过Morris水迷宫实验检测大鼠学习记忆能力,TTC染色检测脑梗死率,HE染色法行海马CA1区和CA3区神经元病理学检查,TUNEL染色法行海马CA1区和CA3区神经元凋亡检查,ELISA法检测海马组织白细胞介素-1β(IL-1β)、IL-8、肿瘤坏死因子-α(TNF-α)含量,Western blot法检测海马组织Janus激酶2/信号转导与转录激活子3(JAK2/STAT3)信号通路相关蛋白相对表达量。结果与Sham组比较,CI/R组学习记忆能力明显降低,脑梗死率明显升高(P<0.05);海马CA1区和CA3区神经元呈现数量减少、间隙增大、空泡样变、核膜核仁边界模糊、炎性细胞浸润等病理改变,凋亡率明显升高(P<0.05);海马组织IL-1β、IL-8、TNF-α含量明显升高(P<0.05);p-JAK2、p-STAT3、高迁移率族蛋白B1(HMGB1)、Bcl-2相关X蛋白(Bax)、激活型半胱氨酸蛋白酶-3(cleaved Caspase-3)相对表达量和p-JAK2/JAK2、p-STAT3/STAT3、Bax/Bcl-2表达比值均明显升高,Bcl-2相对表达量明显降低(P<0.05)。与CI/R组比较,CRO-M组、CRO-H组和NMP组大鼠学习记忆能力显著改善、脑梗死率明显降低(P<0.05);海马CA1区和CA3区神经元病理学改变明显改善、凋亡率明显降低(P<0.05);海马组织IL-1β、IL-8、TNF-α含量明显降低(P<0.05);p-JAK2、p-STAT3、HMGB1、Bax、Cleaved Caspase-3相对表达量和p-JAK2/JAK2、p-STAT3/STAT3、Bax/Bcl-2表达比值均明显降低(P<0.05)。CRO上述作用呈现一定的剂量依赖性,且CRO-H组对CI/R大鼠学习记忆能力、海马CA1区和CA3区神经元病理学改变和凋亡率、炎症因子含量、JAK2/STAT3信号通路相关蛋白表达的影响显著优于NMP组(P<0.05)。结论CRO可能通过抑制JAK2/STAT3信号通路活化,减轻炎症和神经元凋亡,从而对CI/R大鼠海马神经元损伤起到保护作用。

基于JAK/STAT3通路探讨补肾活血方改善单侧输尿管梗阻大鼠炎症和肾纤维化

目的:观察补肾活血方(BSHXF)对单侧输尿管梗阻大鼠(unilateral ureteral obstruction,UUO)肾纤维化和炎症的治疗效果,揭示其抗炎及延缓肾纤维化的机制。方法:32只8周龄雌性SD大鼠根据随机数字表分为假手术组(Sham组)、单侧输尿管梗阻组(UUO组)、补肾活血方低剂量组(BSHXF-L组)和补肾活血方高剂量组(BSHXF-H组)。造模后第2天开始,连续灌胃干预21 d,BSHXF-L组和BSHXF-H组给予相应剂量补肾活血方汤剂,Sham组和UUO组给予与BSHXF组等体积的生理盐水。干预结束后,记录24 h尿量,全自动生化分析仪检测血肌酐、尿素氮、尿蛋白、尿肌酐和肾功能指数;采用HE和Masson染色检测肾脏病理损伤和纤维化程度;通过免疫组化染色分析各组TGF-β和TNF-α;Western blotting检测各组TGF-β_(1)、α-SMA、JAK、STAT3、p-STAT3、TNF-α、Caspase-3和Bax的蛋白表达情况。结果:与UUO模型组相比,BSHXF-L组和BSHXF-H组能明显明显改善UUO大鼠的肾功能,降低大鼠肾组织中TGF-β_(1)和α-SMA蛋白表达水平,抑制JAK、STAT3和p-STAT3表达,减少炎症细胞浸润和纤维组织以及胶原的增生,显著降低大鼠的肾脏指数和TNF-α等炎症指标。结论:补肾活血方可通过抑制JAK/STAT3通路激活,改善UUO大鼠肾功能,减轻其肾脏炎症水平、肾脏指数,延缓肾纤维化进程。