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.

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.

Mechanism of Yanghe Pingchaun granules on airway remodeling in asthmatic rats based on IL-6/JAK2/STAT3 signaling axis

Objective: To investigate the effects of Yanghe Pingchuan Granules on airway remodeling in asthmatic rats, and to explore the mechanism of Interleukin-6/Janus kinase 2/ Signal transducing activator of transcription 3(IL-6/JAK2/STAT3) signal axis. Methods: We separated 42 healthy male SD rats into two groups, a control group (7) and a model group (35).The model group was sensitized with a combination of ovalbumin (OVA) and aluminum hydroxide for 2 weeks, while the control group was given an equal amount of physiological saline.After 2 weeks, the modeling group was randomly divided into Model group, Yanghe Pingchuan Granules high, medium and low dose groups and Dexamethasone group, each group consisted of 7 animals. After 4 weeks, OVA atomization and gavage were used for stimulation and treatment. Yanghe Pingchuan Granules high, middle and low groups were given 15.48, 7.74, 3.87 g∙kg-1 Yanghe Pingchuan Granules daily, dexamethasone group was given 0.0625 mg∙kg-1 dexamethasone daily, and the other groups were given the same amount of normal saline. HE, PAS and Masson staining were used to observe the lung histopathological changes in rats. The levels of interleukin-6, IL-23 and IL-17A were detected by ELISA. The expression levels of JAK-2, P-JAK2, STAT3 and P-STAT3 in lung tissues were detected by Western blot. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the mRNA expression levels of IL-6, JAK2 and STAT3 in rat lung tissue. Results: The lung tissue structure of the model group was severely damaged compared to the control group, accompanied by a great many of inflammatory cell infiltration, goblet cell hyperplasia, subepithelial collagen fiber deposition and airway epithelial thickening were more obvious. The expressions of IL-6, IL- 23 and IL-17A in serum were significantly increased (P<0.01), the protein expression levels of JAK-2, P-JAK2, STAT3 and P-STAT3 and the mRNA expression levels of IL-6, JAK2 and STAT3 in lung tissue were significantly increased (P<0.01);Compared with the model group, inflammatory cell infiltration, goblet cell proliferation, subepithelial collagen fiber deposition and airway epithelial thickening were significantly reduced in each administration group, and the expressions of IL-6, IL-23 and IL-17A in serum were significantly decreased (P< 0.01). The protein expression levels of JAK-2, P-JAK2, STAT3 and P-STAT3 and mRNA expression levels of IL-6, JAK2 and STAT3 in lung tissue were significantly decreased (P<0.01). Conclusion: Yanghe Pingchuan Granules can significantly alleviate airway remodeling in asthmatic rats, and its mechanism may be through inhibiting the IL-6/JAK2/STAT3 signal axis.

Effects of plumbagin on migration and invasion of human hepatoma cell line via JAK2/STAT3 signaling pathway

Objective:To study the effect of plumbagin(PL)on the migration and invasion of human hepatocellular carcinoma(HCC)cells and its possible mechanism.Methods:The cell counting kit(CCK-8)was used to detect the effects of different concentrations of plumbagin on the proliferation of human hepatocellular carcinoma Huh-7 and LM3 cells.The effect of plumbagin on the migration ability of Huh-7 and LM3 cells was detected by scratch test and Transwell migration test,and the effect of on the invasion ability of Huh-7 and LM3 cells was detected by Transwell invasion test.Western Blot was used to detect the expression of E-cadherin,N-cadherin,matrix metalloproteinase-2 and related proteins in JAK2/STAT3 signaling pathway in Huh-7 and LM3 cells.Results:Plumbagin could inhibit the proliferation of Huh-7 and LM3 cells in a time-and concentration-dependent manner.Plumbagin inhibited the migration and invasion of Huh-7 and LM3 cells in a concentration dependent manner,and it can down-regulate the expression of N-cadherin and MMP-2 protein,up-regulate the expression of E-cadherin protein,and inhibit the activation of JAK2/STAT3 signaling pathway.Conclusion:Plumbagin can inhibit the migration and invasion of human hepatocellular carcinoma Huh-7 and LM3 cells,and the molecular mechanism of this process may be related to the inhibition of JAK2/STAT3 signaling pathway activation.

To explore the mechanism of Dahuang Lingxian Formula in relieving inflammatory response of bile duct cells based on IL-6/JAK/STAT3 signaling pathway

Objective:To explore the mechanism of action of Dahuang Lingxian Formula in alleviating the inflammatory response of bile duct cells in LPS-induced intrahepatic bile duct inflammation model rats based on IL-6/JAK/STAT3 signaling pathway.Methods:Fifty SD rats were randomly divided into five groups,blank group,model group,choling tablets(0.5 g/kg),and low and high concentration groups(2.4 g/kg and 4.8 g/kg)of Dahuang Lingxian Formula,ten rats in each group.Except for the blank group,the rats in each group were injected with 1.25 mg/kg LPS at the common bile duct at one time to construct an animal model of intrahepatic bile duct infection.After gavage on day 8,liver tissues were taken from rats at the hepatic hilum,and the histopathological changes of the hepatic hilum and biliary tree were observed by HE staining.The expression levels of serum glutamic alanine transaminase(ALT),glutamic oxalacetic transaminase(AST),malondialdehyde(MDA)and superoxide dismutase(SOD)were measured by biochemical method.The expression levels of interleukin 6(IL-6),Janus protein tyrosine kinase 2(JAK2),signal transducer and activator of transcription 3(STAT3)in rat serum were measured by enzyme-linked immunosorbent assay(ELISA).Protein immunoblotting(WB)and real-time fluorescence quantitative PCR(RT-qPCR)were used to detect the expression levels of IL-6,JAK2,STAT3 protein and mRNA in biliary tree tissues.Results:①Compared with the blank group,the structures such as interlobular bile ducts in the hepatic sinusoids and portal duct area of the model rats were destroyed,and inflammatory cells infiltrated around them.The expression of ALT,AST,MDA,IL-6,JAK2 and STAT3 in the serum increased significantly,the expression level of SOD decreased,and the expression levels of IL-6,JAK2 and STAT3 proteins and mRNA increased.②Compared with the model group,the degree of liver pathological damage in rats in the Chiling Ning tablet group and the low and high concentration groups of Dahuang Lingxian Formula were improved,which could significantly reduce the expression levels of ALT,AST,MDA,IL-6,JAK2,STAT3 and up-regulate SOD in serum,and down-regulate the expression of IL-6,JAK2,STAT3 protein and mRNA,with the best effect in the high concentration group of Dahuang Lingxian Formula.③Compared with the choling tablet group,the rats in the low and high concentration groups of Dahuang Lingxian Formula tended to normalize the degree of liver pathological damage,without obvious inflammatory cell infiltration,and the expression levels of ALT,AST,MDA,IL-6,JAK2,STAT3 and the expression levels of IL-6,JAK2,STAT3 protein and mRNA in serum were reduced,and the expression levels of SOD were increased,with the best effect of Dahuang Lingxian Formula The treatment effect was best in the high concentration group.Conclusion:The mechanism may be related to the down-regulation of IL-6/JAK/STAT3 signaling pathway activation,and the best therapeutic effect was achieved by the high concentration group of Dahuang Lingxian Formula.

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。

Elevated retinol binding protein 4 levels are associated with atherosclerosis in diabetic rats via JAK2/STAT3 signaling pathway

BACKGROUND Atherosclerosis is a major cause of mortality worldwide and is driven by multiple risk factors,including diabetes,which results in an increased atherosclerotic burden,but the precise mechanisms for the occurrence and development of diabetic atheroscerosis have not been fully elucidated.AIM To summarize the potential role of retinol binding protein 4(RBP4) in the pathogenesis of diabetic atheroscerosis,particularly in relation to the RBP4-Janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3)signaling pathway.METHODS Male Wistar rats were randomly divided into three groups,including a control group(NC group),diabetic rat group(DM group),and diabetic atherosclerotic rat group(DA group).The contents of total cholesterol(TC), high-density lipoprotein cholesterol(HDL-c), triglycerides(TG), low-density lipoprotein cholesterol(LDLc), fasting insulin(FINS),fasting plasma glucose,and hemoglobin A1 c(HbA1 c)were measured.Moreover,the adipose and serum levels of RBP4,along with the expression levels of JAK2, phosphorylated JAK2(p-JAK2), STAT3,phosphorylated STAT3(p-STAT3), B-cell lymphoma-2(Bcl-2), and Cyclin D1 in aortic tissues were also measured.Besides,homeostasis model assessment of insulin resistance(HOMA-IR) and atherogenic indexes(AI) were calculated.RESULTS Compared with the NC and DM groups,the levels LDL-c,TG,TC,FINS,HOMAIR,RBP4,and AI were upregulated,whereas that of HDL-c was downregulated in the DA group(P <0.05);the mRNA levels of JAK2,STAT3,Cyclin D1,and Bcl-2 in the DA group were significantly increased compared with the NC group and the DM group;P-JAK2,p-JAK2/JAK2 ratio,p-STAT3,p-STAT3/STAT3 ratio,Cyclin D1,and Bcl-2 at protein levels were significantly upregulated in the DA group compared with the NC group and DM group.In addition,as shown by Pearson analysis,serum RBP4 had a positive correlation with TG,TC,LDL-c,FINS,HbA1 C,p-JAK2,p-STAT3,Bcl-2,Cyclin D1,AI,and HOMA-IR but a negative correlation with HDL-c.In addition,multivariable logistic regression analysis showed that serum RBP4,p-JAK2,p-STAT3,and LDL-c were predictors of the presence of diabetic atherosclerosis.CONCLUSION RBP4 could be involved in the initiation or progression of diabetic atherosclerosis by regulating the JAK2/STAT3 signaling pathway.

3-epi-bufotalin suppresses the proliferation in colorectal cancer cells through the inhibition of the JAK1/STAT3 signaling pathway

Traditional Chinese medicine(TCM)has been increasingly employed in the last decades in China for both preventing and treating a variety of cancers.3-epi-bufotalin is an active ingredient of TCM“Chanpi”with anti-tumor potential.However,the effect and mechanism of 3-epi-bufotalin on colorectal cancers were not well disclosed.The present study demonstrated that 3-epi-bufotalin could reduce viability,trigger apoptosis,and block the cell cycle at the G2/M stage in colorectal cancer cell lines HT29,RKO,and COLO205 in vitro.Moreover,3-epi-bufotalin inhibited the JAK1/STAT3 signaling pathway.These results indicated the anti-proliferation ability of 3-epi-bufotalin in colorectal cancer cells.

中药调控JAK/STAT信号通路干预心肌缺血再灌注损伤作用机制研究进展

急性心肌梗死是临床常见的心血管急危重症,早期再灌注是治疗急性心肌梗死的常规有效方法,但血液供应的恢复可能造成心肌缺血再灌注损伤(MI/RI),从而加重心肌损伤。近年来研究发现,中药在干预MI/RI方面具有多成分、多途径、多靶点的独特优势。Janus酪氨酸蛋白激酶/信号转导及转录激活因子(JAK/STAT)信号通路与MI/RI密切相关,通过调控炎症、氧化应激、细胞增殖、分化、凋亡等作用减轻MI/RI进程。本文就JAK/STAT信号通路在MI/RI中的作用机制及靶向调控该通路的中药研究进行综述,以期为MI/RI的防治及药物研发提供参考。

基于JAK/STAT信号通路探讨PRELID1表达在胃癌恶性生物学行为中的作用

目的探讨相关进化和淋巴兴趣域蛋白1(PRELID1)在胃癌组织中的表达及对预后的影响,并分析其影响胃癌细胞增殖和侵袭能力的机制。方法利用TCGA数据库和111例胃癌患者的临床数据分析PRELID1在胃癌组织中的表达,并分析PRELID1表达与临床病理特征的关系及对预后的影响。生物信息学技术预测PRELID1的生物学功能,并进一步采用体外和体内实验验证。体外实验检测慢病毒调控胃癌细胞系(MGC803)中PRELID1的表达,并观察其对胃癌细胞增殖、迁移和侵袭的影响。利用裸鼠皮下成瘤体内实验观察PRELID1表达对胃癌组织生长的影响。结果PRELID1在胃癌组织中的表达显著高于癌旁组织(P<0.001)且与Ki67增殖指数呈正相关(P<0.001)。Cox回归模型分析显示,PRELID1高表达是影响胃癌患者根治术后5年生存率的独立危险因素(HR=2.336;95%CI=1.354~4.029)。基因富集结果显示,PRELID1的功能与细胞增殖和JAK/STAT信号有关。CCK-8和Transwell实验发现上调PRELID1表达可促进胃癌细胞的增殖(P=0.016)、迁移(P=0.016)和侵袭(P=0.025),下调其表达则抑制胃癌细胞的增殖(P=0.026)、迁移(P=0.048)和侵袭(P=0.029);裸鼠皮下成瘤实验发现上调PRELID1表达可促进胃癌组织的生长(P=0.047),下调其表达结果相反(P=0.005)。Western blot法检测结果显示,上调PRELID1表达可促进胃癌细胞和胃癌组织中JAK和STAT蛋白的表达(P均<0.05),下调则抑制(P均<0.05)。结论PRELID1在胃癌组织中呈高表达且与预后不良相关,其可能通过上调JAK/STAT信号调控胃癌细胞的增殖、迁移和侵袭。

中药调控JAK/STAT信号通路治疗骨关节炎的研究进展

骨关节炎(osteoarthritis,OA)涵盖了多部位病变,包括关节软骨、韧带、关节囊和滑膜组织等病变,产生骨赘和骨硬化等现象,致使关节软骨受损。OA主要表现为关节间隙狭窄、滑膜炎症、软骨重塑和分解,伴随着关节周围肌肉组织发生改变。OA主要发生在膝、髋、手和脊柱等部位,临床表现以关节的慢性疼痛、局部肿胀僵硬伴活动受限,甚至影响关节功能活动,影响患者日常活动[1]。

JAK/STAT信号通路在类风湿关节炎致病机制及治疗靶点中的作用进展

类风湿关节炎(RA)是一种以滑膜炎及骨破坏为特征的全身炎症性自身免疫性疾病,若未及时治疗,最终会发展为关节畸形、功能障碍,甚至残疾。Janus激酶(JAK)转录活化子(STAT)信号通路在RA的发生发展中扮演关键角色,针对该通路的治疗靶点使RA疾病缓解成为现实,故成为近年来研究的热点。本文就JAK/STAT信号通路的结构与功能,对该通路参与RA滑膜炎症、软骨及骨侵蚀的作用机制进行阐释,总结基础实验和临床药物对该通路治疗靶点的最新研究成果,重点对目前全球批准的14种JAK抑制剂最新研究现状进行综述,为更多RA治疗药物的研发提供新思路。

Value of spiral CT perfusion parameters for evaluating acute pancreatitis and their correlation with inflammatory factor and JAK2/STAT3 signaling pathway

Objective: To study the value of spiral CT perfusion parameters for evaluating acute pancreatitis and their correlation with inflammatory factor and JAK2/STAT3 signaling pathway. Methods: Patients with acute pancreatitis and patients with pancreatic trauma who underwent surgical resection in Liaocheng Dongchangfu People's Hospital between May 2014 and March 2017 were selected and enrolled in the AP group and the control group of the research respectively;spiral CT perfusion scanning was conducted before surgery to measure the blood flow (BF), blood volume (BV), and mean transit time (MTT), and the serum was collected to determine the contents of inflammatory factors;pancreatitis tissue and normal pancreatic tissue were collected after surgical resection to determine the expression of JAK2/STAT3 signal molecules. Results: pancreatic tissue BF and BV levels of AP group were significantly lower than those of control group while MTT level was not different from that of control group;CRP, PCT, HMGB-1, Ghrelin and sTREM-1 contents in serum as well as JAK2, STAT3, Bcl-2 and Bcl-xL mRNA expression in pancreatic tissue of AP group were significantly higher than those of control group and negatively correlated with BF and BV levels in pancreatic tissue. Conclusion: Spiral CT perfusion parameters BF and BV can reflect the microcirculatory disorder of acute pancreatitis and are associated with the increased secretion of inflammatory factors and the activation of JAK2/STAT3 signaling pathway in the course of disease.

lncRNA RMRP通过JAK/STAT信号通路在子宫内膜癌细胞增殖和凋亡中的作用研究

目的 探讨长链非编码RNA RMRP(lncRNA RMRP)在子宫内膜癌中的生物学功能及主要分子机制。方法 收集在我院接受手术治疗的30例子宫内膜癌患者的癌组织和癌旁组织标本,RT-qPCR法检测lncRNA RMRP在子宫内膜癌组织和癌旁组织、HESC细胞和HEC-1-A细胞中的表达。体外培养子宫内膜癌细胞系HEC-1-A,将空载体、pcDNA-RMRP、NC-siRNA、RMRPsiRNA、NCmimic、miR-580-3pmimic、pcDNA-RMRP+NCmimic、pcDNA-RMRP+miR-580-3pmimic、RMRP-siRNA+空载体、RMRPsiRNA+pcDNA-JAK2、NC inhibitor、miR-580-3p inhibitor分别转染至HEC-1-A细胞中,作为空载体组、pcDNA-RMRP组、NC-siRNA组、RMRP-siRNA组、NC mimic组、miR-580-3p mimic组、pcDNA-RMRP+NC mimic组、pcDNA-RMRP+miR-580-3p mimic组、RMRP-siRNA+空载体组、RMRP-siRNA+pcDNA-JAK2组、NC inhibitor组、miR-580-3p inhibitor组。RT-qPCR检测lncRNA RMRP、miR-580-3p在细胞中的表达;CCK-8法检测细胞增殖率;流式细胞术检测细胞凋亡率;生物信息学软件和双荧光素酶报告基因实验分别预测和验证miR-580-3p与lncRNA RMRP、JAK2的靶向互作关系;Western blot检测JAK/STAT信号通路蛋白表达。结果 与癌旁组织相比,lncRNA RMRP在子宫内膜癌组织中均显著高表达(P<0.05)。与HESC细胞比较,lncRNA RMRP在HEC-1-A细胞中的表达显著升高(P<0.05)。pcDNA-RMRP可显著促进细胞增殖,抑制细胞凋亡,而RMRP-siRNA可显著抑制细胞增殖,促进细胞凋亡,差异均有统计学意义(P<0.05)。miR-580-3p是lncRNA RMRP的下游靶miRNA,lncRNA RMRP可负向调控miR-580-3p的表达。JAK2是miR-580-3p的下游靶基因,miR-580-3p可负向调控JAK2蛋白的表达。pcDNA-RMRP可显著增加细胞中JAK2、p-JAK2和p-STAT3蛋白水平,而pcDNA-RMRP与miR-580-3p mimic共转染后,细胞中JAK2、p-JAK2、p-STAT3蛋白水平显著降低,差异均有统计学意义(P<0.05)。RMRP-siRNA可显著降低细胞中JAK2、p-JAK2、p-STAT3蛋白水平,而RMRP-siRNA与pcDNA-JAK2共转染后,细胞中JAK2、p-JAK2、p-STAT3蛋白水平显著升高,差异均有统计学意义(P<0.05)。此外,RMRP-siRNA与pcDNA-JAK2共转染后,细胞增殖率升高,细胞凋亡率降低,差异均有统计学意义(P<0.05)。结论 敲低lncRNA RMRP通过调控JAK2/STAT3信号通路抑制子宫内膜癌细胞增殖,促进细胞凋亡,可能成为子宫内膜癌潜在的治疗靶点。

LAIR-1通过阻断JAK2 V617F突变的人HEL细胞JAK/STAT和PI3K/AKT/mTOR信号通路抑制其增殖并促进其凋亡

目的研究人白细胞相关免疫球蛋白样受体1(LAIR-1)对Janus激酶2(JAK2)V617F突变的人急性髓系白血病HEL细胞JAK/信号转导子与转录激活子(STAT)和磷脂酰肌醇3激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白(PI3K/AKT/mTOR)信号通路的调节作用,以及对细胞增殖和凋亡的影响。方法采用反转录PCR和基因测序鉴定JAK2 V617F突变;应用免疫共沉淀和Western blot法鉴定LAIR-1募集的蛋白酪氨酸磷酸酶(PTP)种类;采用CCK-8法检测HEL细胞的增殖;采用异硫氰酸荧光素标记的膜联素Ⅴ/碘化丙啶(annexinⅤ-FITC/PI)双标记结合流式细胞术检测HEL细胞的凋亡率;采用Western blot法检测JAK/STAT和PI3K/AKT/mTOR通路蛋白酪氨酸磷酸化水平及细胞周期蛋白D1(cyclin D1)、Bcl2相关X蛋白(BAX)和B细胞淋巴瘤因子2(Bcl2)的蛋白表达。结果在JAK2 V617F突变的HEL细胞中,LAIR-1与其配体胶原蛋白结合后可募集含Src同源域2磷酸酶2(SHP-2);LAIR-1可以下调HEL细胞JAK2、STAT1、STAT3、STAT5、AKT和mTOR的蛋白酪氨酸磷酸化水平,并能够显著抑制cyclin D1和Bcl2的表达,而对BAX的表达水平未见显著影响;LAIR-1能够明显抑制HEL细胞的增殖,促进HEL细胞凋亡。结论在JAK2 V617F突变的人白血病HEL细胞中,LAIR-1可通过募集SHP-2抑制JAK/STAT和PI3K/AKT/mTOR信号通路的活化,进而抑制HEL细胞的增殖,促进细胞凋亡。

JAK/STAT通路介导椎间盘髓核细胞凋亡的研究进展

椎间盘退变(Intervertebral disc degeneration, IDD)是一种与年龄相关的退行性疾病。近年来,IDD的发病率逐年升高,已成为全球关注的公共卫生问题。髓核细胞作为椎间盘内的主要细胞,其凋亡是椎间盘发生退变的重要病理基础之一。研究表明Janus激酶(Janus kinase, JAK)/信号转导子及转录激活因子(Signal transducer and activator of transcriptor, STAT)信号途径与髓核细胞凋亡过程联系密切。本文就JAK/STAT信号通路介导的椎间盘髓核细胞凋亡的研究进展及以JAK/STAT信号通路为潜在治疗靶点进行综述,以期为IDD的治疗提供参考。

Morroniside ameliorates lipopolysaccharide-induced inflammatory damage in iris pigment epithelial cells through inhibition of TLR4/JAK2/STAT3 pathway

AIM:To investigate the effect of morroniside(Mor)on lipopolysaccharide(LPS)-treated iris pigment epithelial cells(IPE).METHODS:IPE cells were induced by LPS and treated with Mor.Cell proliferation was detected by cell counting kit(CCK)-8,apoptosis was detected by flow cytometry,the levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-6,and IL-8 were measured by enzyme-linked immunosorbent assay(ELISA)kits,and the protein expression of TLR4,JAK2,p-JAK2,STAT3,and p-STAT3 was analyzed by Western blotting.In addition,overexpression of TLR4 and Mor treatment of LPS-stimulated IPE cells were also tested for the above indices.RESULTS:Mor effectively promoted the proliferation and inhibited the apoptosis of LPS-treated IPE cells.In addition,Mor significantly reduced the levels of TNF-α,IL-6,and IL-8 and significantly inhibited the expression of TLR4,p-JAK2,and p-STAT3 in LPS-treated IPE cells.The effect of Mor on LPS-treated IPE cells was markedly attenuated after overexpression of TLR4.CONCLUSION:These findings suggest that Mor may ameliorate LPS-induced inflammatory damage and apoptosis in IPE through inhibition of TLR4/JAK2/STAT3 pathway.

Differential regulation of JAK/STAT-signaling in patients with ulcerative colitis and Crohn’s disease

In 2018,the pan-Janus kinase(JAK)inhibitor tofacitinib was launched for the treatment of ulcerative colitis(UC).Although tofacitinib has proven efficacious in patients with active UC,it failed in patients with Crohn’s disease(CD).This finding strongly hints at a different contribution of JAK signaling in both entities.Here,we review the current knowledge on the interplay between the JAK/signal transducer and activator of transcription(STAT)pathway and inflammatory bowel diseases(IBD).In particular,we provide a detailed overview of the differences and similarities of JAK/STAT-signaling in UC and CD,highlight the impact of the JAK/STAT pathway in experimental colitis models and summarize the published evidence on JAK/STAT-signaling in immune cells of IBD as well as the genetic association between the JAK/STAT pathway and IBD.Finally,we describe novel treatment strategies targeting JAK/STAT inhibition in UC and CD and comment on the limitations and challenges of the new drug class.

JAK/STAT signaling regulates tissue outgrowth and male germline stem cell fate in Drosophila

In multicellular organisms, biological activities are regulated by cell signaling. The various signal transduction path- ways regulate cell fate, proliferation, migration, and polarity. Miscoordination of the communicative signals will lead to disasters like cancer and other fatal diseases. The JAK/STAT signal transduction pathway is one of the pathways, which was first identified in vertebrates and is highly conserved throughout evolution. Studying the JAK/STAT signal transduc- tion pathway in Drosophila provides an excellent opportunity to understand the molecular mechanism of the cell regu- lation during development and tumor formation. In this review, we discuss the general overview of JAK/STAT signaling in Drosophila with respect to its functions in the eye development and stem cell fate determination.

HBP1 inhibits chicken preadipocyte differentiation by activating the STAT3 signaling via directly enhancing JAK2 expression

Obesity presents a serious threat to human health and broiler performance.The expansion of adipose tissue is mainly regulated by the differentiation of preadipocytes.The differentiation of preadipocytes is a complex biological process regulated by a variety of transcription factors and signaling pathways.Previous studies have shown that the transcription factor HMG-box protein 1(HBP1)can regulate the differentiation of mouse 3T3-L1 preadipocytes by activating the Wnt/β-catenin signaling pathway.However,it is unclear whether HBP1 involved in chicken preadipocyte differentiation and which signaling pathways it regulates.The aim of the current study was to explore the biological function and molecular regulatory mechanism of HBP1 in the differentiation of chicken preadipocytes.The expression patterns of chicken HBP1 in abdominal adipose tissue and during preadipocyte differentiation were analyzed by RT-qPCR and Western blot.The preadipocyte stably overexpressing HBP1 or knockout HBP1 and their control cell line were used to analyze the effect of HBP1 on preadipocyte differentiation by oil red O staining,RT-qPCR and Western blot.Cignal 45-Pathway Reporter Array was used to screen the signal pathways that HBP1 regulates in the differentiation of chicken preadipocytes.Chemical inhibitor and siRNA for signal transducer and activator of transcription 3(STAT3)were used to analyze the effect of STAT3 on preadipocyte differentiation.The preadipocyte stably overexpressing HBP1 was transfected by the siRNA of STAT3 or treated with a chemical inhibitor of STAT3 for the rescue experiment.The results of gene expression analysis showed that the expression of HBP1 was related to abdominal fat deposition and preadipocyte differentiation in chickens.The results of function gain and loss experiments indicated that overexpression/knockout of HBP1 in chicken preadipocytes could inhibit/promote(P<0.05)lipid droplet deposition and the expression of adipogenesis-related genes.Mechanismlly,HBP1 activates(P<0.05)the signal transducer and activator of transcription 3(STAT3)signaling pathway by targeting janus kinase 2(JAK2)transcription.The results of functional rescue experiments indicated that STAT3 signaling mediated the regulation of HBP1 on chicken preadipocyte differentiation.In conclusion,HBP1 inhibits chicken preadipocyte differentiation by activating the STAT3 signaling pathway via directly enhancing JAK2 expression.Our findings provided new insights for further analysis of the molecular genetic basis of chicken adipose tissue growth and development.