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.

SEMA3F通过CXCR4/JAK2/STAT3通路抑制子宫内膜癌细胞增殖、侵袭、迁移及血管生成

目的:探究信号素3F(semaphorin-3F,SEMA3F)通过趋化因子受体4(C-X-C motif chemokine receptor 4,CXCR4)/JANUS激酶2(janus kinase 2,JAK2)/信号转导及转录激活蛋白3(signal transducer and activator of transcription 3,STAT3)通路对子宫内膜癌细胞增殖、侵袭、迁移及血管生成的调控作用。方法:体外培养人子宫内膜基质细胞系、人子宫内膜癌细胞(Ishikawa、KLE、RL95-2、HEC-1-A)及人脐静脉血管内皮细胞(human umbilical vein endothelial cells,HUVEC)。构建含有SEMA3F重组质粒的慢病毒载体并转染至Ishikawa细胞,分为对照组(Control)、过表达对照组(Ov-NC)、SEMA3F过表达组(Ov-SEMA3F);将含SEMA3F重组质粒的慢病毒载体与构建的含CXCR4重组质粒的慢病毒载体或STAT3激动剂(Colivelin)共转染至Ishikawa细胞,分为4组:Control组、Ov-SEMA3F组、Ov-SEMA3F+Ov-CXCR4组、Ov-SEMA3F+Colivelin组。实时荧光定量PCR(quantitative real-time PCR,RT-qPCR)、蛋白质印迹法(Western blot)检测SEMA3F、CXCR4表达水平;EdU染色检测细胞增殖;划痕和Transwell实验分别检测细胞迁移和侵袭能力;Transwell和小管形成实验分别检测Ishikawa细胞上清作用下HUVEC迁移和子宫内膜癌细胞小管形成能力;Western blot分析增殖和转移相关蛋白及CXCR4/JAK2/STAT3通路相关蛋白表达水平。结果:SEMA3F在子宫内膜癌细胞中表达降低(均P<0.01),SEMA3F过表达可抑制细胞增殖、迁移、侵袭和血管生成(均P<0.001)。SEMA3F过表达可抑制CXCR4/JAK2/STAT3信号通路(P<0.001),CXCR4过表达或STAT3激动剂(Colivelin)可逆转SEMA3F对Ishikawa细胞增殖、迁移、侵袭及Ishikawa细胞上清作用下HUVEC细胞迁移、血管生成能力的抑制作用(均P<0.05)。结论:SEMA3F可通过使CXCR4/JAK2/STAT3通路失活进而抑制子宫内膜癌细胞增殖、侵袭、迁移及血管生成。

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.

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.

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.

JAK2-STAT3/STAT5信号通路与儿童食物过敏CD4^+T淋巴细胞亚群变化的相关性研究

目的:探讨JAK2-STAT3/STAT5信号通路与儿童食物过敏CD4^+T淋巴细胞亚群变化的相关性。方法:收集2015年1月至2017年3月来我院就诊的食物过敏患儿78例,为过敏组;同期收集来我院体检的健康儿童78例,为对照组。并根据儿童年龄将过敏组和对照组分为A、B、C、D四组,其中A组≤1岁,B组1~3岁(不包括1岁),C组3~6岁(不包括3岁),D组6~11岁(不包括6岁)。采用流式细胞仪检测CD4^+T淋巴细胞亚群Th1、Th2、Th17和调节性T淋巴细胞(Treg)百分比,同时采用q PCR检测外周血单核细胞JAK2、STAT3、STAT5的mRNA水平,分析两组儿童上述CD4^+T淋巴细胞亚群和JAK2、STAT3、STAT5表达水平的差异,并进一步分析JAK2-STAT3/STAT5信号通路和CD4^+T淋巴细胞亚群的相关性。结果:对照组A组、B组、C组儿童Th1、Treg淋巴细胞百分比及Th1/Th2、Treg/Th17比值均明显高于过敏组患儿A组、B组、C组,而Th2、Th17淋巴细胞百分比均小于过敏组所对应的同年龄段患儿,随儿童年龄增大,两组间差异均明显减小,但差异均有统计学意义(P<0.05)。对照组D组儿童和过敏组D组儿童上述指标差异均无统计学意义(P>0.05)。分别针对A、B、C三组患儿JAK2、STAT3、STAT5和CD4^+T淋巴细胞亚群水平进行相关性比较,Th1、Treg、Th1/Th2、Treg/Th17的相关性系数均<0(P<0.05),而Th2、Th17的相关性系数均>0(P<0.05),且上述三组的相关系数绝对值逐次减小。上述信号通路分子与CD4^+T淋巴细胞亚群水平在D组均无明显相关性。结论:低龄食物过敏儿童的JAK2-STAT3/STAT5信号通路存在明显活化和CD4^+T淋巴细胞亚群改变现象,JAK2-STAT3/STAT5信号通路可有效调控CD4^+T淋巴细胞亚群的分布,并伴随患儿年龄增大,上述作用相关性减弱并消失。

Hepatocellular carcinoma-derived exosomal miRNA-761 regulates the tumor microenvironment by targeting the SOCS2/JAK2/STAT3 pathway

BACKGROUND:Exosomes and exosomal microRNAs have been implicated in tumor occurrence and metastasis.Our previous study showed that microRNA-761(miR-761)is overexpressed in hepatocellular carcinoma(HCC)tissues and that its inhibition affects mitochondrial function and inhibits HCC metastasis.The mechanism by which exosomal miR-761 modulates the tumor microenvironment has not been elucidated.METHODS:Exosomal miR-761 was detected in six cell lines.Cell counting kit-8(CCK-8)and transwell migration assays were performed to determine the function of exosomal miR-761 in HCC cells.The luciferase reporter assay was used to analyze miR-761 target genes in normal fi broblasts(NFs).The inhibitors AZD1480 and C188-9 were employed to determine the role of the Janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3)signaling pathway in the transformation of cancer-associated fi broblasts(CAFs).RESULTS:In this study,we characterized the mechanism by which miR-761 reprogrammed the tumor microenvironment.We found that HCC-derived exosomal miR-761 was taken up by NFs.Moreover,HCC exosomes aff ected the tumor microenvironment by activating NFs via suppressor of cytokine signaling 2(SOCS2)and the JAK2/STAT3 signaling pathway.CONCLUSIONS:These results demonstrated that exosomal miR-761 modulated the tumor microenvironment via SOCS2/JAK2/STAT3 pathway-dependent activation of CAFs.Our fi ndings may inspire new strategies for HCC prevention and therapy.

LncRNA BCAR4调控JAK2/STAT3信号通路对食管癌细胞侵袭、迁移、炎症和凋亡的影响

目的:探讨长链非编码RNA(lncRNA)乳腺癌抗雌激素抵抗4(BCAR4)介导JAK激酶2/信号转导与转录因子3(JAK2/STAT3)信号通路对食管癌细胞迁移、侵袭、凋亡和炎症因子分泌的影响。方法:采用实时荧光定量PCR(qRT-PCR)法检测BCAR4在食管癌细胞中的表达。在食管癌EC9706细胞中转染BCAR4小干扰RNA(si-BCAR4),Transwell和流式细胞术分别检测细胞迁移、侵袭和凋亡,Western blot法检测白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、磷酸化JAK2(pJAK2)、JAK2、磷酸化STAT3(pSTAT3)、STAT3表达。使用JAK2/STAT3信号通路激活剂Colivelin处理EC9706细胞,观察其在BCAR4沉默诱导的食管癌细胞迁移、侵袭、凋亡和炎症因子表达中的作用。结果:与人正常食管鳞状上皮Het-1A细胞比较,食管癌细胞TE-10、OE19、EC9706中BCAR4表达量明显升高(P<0.05)。沉默BCAR4的表达可明显降低EC9706细胞迁移数、侵袭数以及IL-1β、TNF-α、IL-6、pJAK2和pSTAT3表达,并显著提高细胞凋亡率(P<0.05),而对JAK2、STAT3蛋白水平无显著影响(P>0.05)。激活JAK2/STAT3通路逆转了BCAR4沉默对EC9706细胞迁移、侵袭和炎症因子表达的抑制和对细胞凋亡的诱导。结论:LncRNA BCAR4沉默通过调控JAK2/STAT3信号通路活性,抑制食管癌细胞的迁移、侵袭和炎症因子的分泌,并促进细胞凋亡。

Inhibition of cerebral ischemia/reperfusion injuryinduced apoptosis:nicotiflorin and JAK2/STAT3 pathway

Nicotiflorin is a flavonoid extracted from Carthamus tinctorius.Previous studies have shown its cerebral protective effect,but the mechanism is undefined.In this study,we aimed to determine whether nicotiflorin protects against cerebral ischemia/reperfusion injury-induced apoptosis through the JAK2/STAT3 pathway.The cerebral ischemia/reperfusion injury model was established by middle cerebral artery occlusion/reperfusion.Nicotiflorin（10 mg/kg） was administered by tail vein injection.Cell apoptosis in the ischemic cerebral cortex was examined by hematoxylin-eosin staining and terminal deoxynucleotidyl transferase d UTP nick end labeling assay.Bcl-2 and Bax expression levels in ischemic cerebral cortex were examined by immunohistochemial staining.Additionally,p-JAK2,p-STAT3,Bcl-2,Bax,and caspase-3 levels in ischemic cerebral cortex were examined by western blot assay.Nicotiflorin altered the shape and structure of injured neurons,decreased the number of apoptotic cells,down-regulates expression of p-JAK2,p-STAT3,caspase-3,and Bax,decreased Bax immunoredactivity,and increased Bcl-2 protein expression and immunoreactivity.These results suggest that nicotiflorin protects against cerebral ischemia/reperfusion injury-induced apoptosis via the JAK2/STAT3 pathway.

甲状腺乳头状癌组织中USP33和CXCR4表达及其与JAK3、STAT3信号分子的关系

目的探讨PTC组织中USP33和CXCR4表达及其与JAK3、STAT3信号分子的关系。方法采用免疫组化染色EnVision法检测甲状腺乳头状癌(papillary thyroid carcinoma,PTC)组织中USP33及CXCR4的表达;采用Western blot技术检测6例PTC组织及癌旁组织中USP33的表达;采用qRT-PCR技术检测6例PTC组织及癌旁组织中USP33、CXCR4、JAK3及STAT3的表达。结果(1)免疫组化:USP33阳性表达位于细胞质,在肿瘤体积较大患者的PTC组织中表达量下降(P<0.05),CXCR4阳性表达位于细胞质,表达水平与远处转移、TNM分期有关(P<0.001);(2)Western blot:癌旁组织中USP33的表达水平高于PTC组织(P<0.05);(3)qRT-PCR:癌旁组织中USP33 mRNA在的表达略高于PTC组织(P>0.05),CXCR4 mRNA、JAK3 mRNA在PTC组织中的表达明显高于癌旁组织(P<0.05)。在PTC组织中,USP33及CXCR4蛋白表达的弱阳性率分别为42%,3.2%;中度阳性率分别为20%,38.7%;强阳性率为28%,32.3%。结论USP33、CXCR4和JAK3信号分子可能参与了PTC的发生、发展。

IL-2通过Jak3-Stat5通路促进巨噬细胞M1极化

目的探究IL-2在调控巨噬细胞极化分型方面的作用及机制。方法重组小鼠白介素-2(IL-2)刺激处于M0期的小鼠单核巨噬细胞RAW 264.7,同时以IL-4作为对照。Real-time PCR和Western blot检测M1型和M2型标志分子的表达;流式细胞术检测M1型和M2型巨噬细胞的百分比;Western blot检测Jak3和Stat5活化水平。结果经IL-2刺激后,处于M0的RAW 264.7细胞显著上调表达M1型标记分子,如IL-1β、IL-12、TNF-α和i NOS等;IL-2使巨噬细胞中M1型的比例由3.2%上升到24.6%,同时Jak3和Stat5分子的磷酸化水平显著提高。结论 IL-2具有促进巨噬细胞由M0向M1极化的作用,且该作用可能是通过Jak3-Stat5通路实现。

miR-140-3p通过靶向趋化因子受体4和抑制JAK2/STAT3通路减轻缺氧复氧诱导的心肌细胞损伤

[目的]探讨miR-140-3p对缺氧复氧(H/R)诱导的心肌细胞损伤的影响及其机制。[方法]构建体外心肌细胞H/R模型,使用miR-140-3p mimics、趋化因子受体4(CXCR4)过表达质粒转染H9c2细胞。噻唑蓝法检测细胞活性;流式细胞术检测细胞凋亡;反转录聚合酶链反应和Western blot检测细胞中miR-140-3p、CXCR4、Janus蛋白酪氨酸激酶2(JAK2)/信号转导和转录激活因子3(STAT3)通路的激活以及凋亡相关蛋白的表达。双荧光素酶报告基因实验验证miR-140-3p与CXCR4的靶向关系。相应试剂盒检测乳酸脱氢酶(LDH)的活性和炎症因子、活性氧(ROS)的水平。[结果]体外H/R可抑制miR-140-3p的表达,上调CXCR4表达和JAK2/STAT3通路的磷酸化,诱导H9c2细胞凋亡而抑制H9c2细胞增殖,促进炎症因子和ROS的释放并上调LDH的活性。miR-140-3p可以通过靶向CXCR43′UTR抑制CXCR4表达,从而抑制JAK2/STAT3通路的磷酸化激活,抑制炎症因子和ROS的释放,下调LDH的活性,促进H9c2细胞增殖而抑制其凋亡。[结论]miR-140-3p可能通过靶向抑制CXCR4而抑制JAK2/STAT3通路,从而减轻缺血再灌注诱导的心肌细胞损伤。

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.

结合网络药理学从JAK2/STAT3信号通路探讨麻杏甘石汤抗流感病毒的效应机制

目的结合网络药理学探索JAK2/STAT3信号通路在A型流感病毒感染所致肺组织损伤中的影响,并探讨麻杏甘石汤对该通路的干预作用。方法通过网络药理学方法筛选麻杏甘石汤作用于流感病毒的潜在靶点所富集的信号通路;以BLAB/c小鼠为研究对象,用A型流感病毒进行滴鼻感染,设置正常对照组、模型对照组、奥司他韦组、抗病毒颗粒组和麻杏甘石汤组,给予相应药物3、7 d后,处理动物。常规法检测小鼠体质量并计算肺指数,观察肺组织病理变化,RT-PCR法检测肺组织中JAK2、STAT3、IL^(-1)β、IL-4 mRNA表达水平,Western blot法或ELISA法检测肺组织中JAK2、STAT3、IL^(-1)β和IL-4的蛋白的表达水平;利用AutoDock Vina软件对STAT3与靶点化合物进行分子对接。结果麻杏甘石汤主要活性成分与流感病毒有110个交集靶点,富集于170条信号通路;麻杏甘石汤治疗组小鼠体质量增加,肺组织病理损伤减轻,肺指数下调,肺组织中JAK2、STAT3、IL-1βmRNA和蛋白的表达水平下调,IL-4 mRNA和蛋白的表达水平上调;STAT3与麻杏甘石汤中活性化合物甘草查尔酮A有较好的结合活性。结论麻杏甘石汤能有效地减轻肺部病理变化、调节免疫平衡,其可能的作用机制是通过抑制JAK2/STAT3信号通路的激活而缓解A型流感病毒感染所致的肺损伤。

钙激活的氯离子通道A4通过抑制JAK激酶2/信号转导及转录激活蛋白3信号通路对食管癌细胞增殖、迁移及侵袭的影响

目的探讨钙激活的氯离子通道A4(CLCA4)对食管癌细胞增殖、迁移、侵袭及JAK激酶2/信号转导及转录激活蛋白3(JAK2/STAT3)信号通路的影响。方法实时荧光定量逆转录聚合酶链反应(qRT-PCR)与蛋白质印迹法(Western blotting)分别检测正常人食管鳞状上皮细胞与人食管癌细胞中CLCA4的表达;将合成的CLCA4过表达载体及其对照分别转染至食管癌细胞Eca109,分别记作CLCA4过表达(pcDNA-CLCA4)组、CLCA4阴性对照(pcDNA-NC)组,并将未转染的细胞作为阴性对照(NC)组。四甲基偶氮唑盐微量酶反应比色法(MTT法)检测细胞增殖能力;细胞迁移实验(Transwell)检测细胞迁移及侵袭能力。JAK2/STAT3信号通路激活剂p-JAK2多肽对细胞增殖、迁移及侵袭的影响。Western blotting检测细胞周期蛋白D1(Cy⁃clinD1)、依赖性激酶抑制因子(P21)、基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)、JAK2、STAT3、磷酸化JAK激酶2(p-JAK2)、磷酸化信号转导及转录激活蛋白3(p-STAT3)的表达水平。结果与Het-1A相比,人食管癌细胞KYSE170、Eca109、TE10中CLCA4 mRNA及蛋白表达水平降低(P<0.05);与pcDNA-NC组比较,pcDNA-CLCA4组细胞存活率显著降低[(52.16±11.41)%比(99.57±13.49)%,P<0.05],迁移细胞数[(56.47±10.03)%比(112.49±13.52)%]与侵袭细胞数[(63.43±9.87)%比(123.47±16.58)%]减少(P<0.05),CyclinD1、MMP-2、MMP-9、p-JAK2、p-STAT3的表达水平降低(P<0.05),P21的表达水平升高(P<0.05);激活JAK2/STAT3信号通路可逆转CLCA4过表达对Eca109细胞增殖、迁移及侵袭的抑制作用。结论CL⁃CA4过表达可抑制食管癌细胞增殖、迁移及侵袭,其作用机制可能与抑制JAK2/STAT3信号通路活化有关。

温阳通脉方通过JAK2/STAT3信号通路保护大鼠心肌缺血再灌注损伤

目的探讨温阳通脉方是否通过Janus蛋白酪氨酸激酶2(JAK2)/信号转导和转录激活因子3(STAT3)信号转导通路,调节水通道蛋白(AQP)的表达,发挥对缺血再灌注心肌组织的保护作用。方法将30只雄性SD大鼠随机分为5组,每组6只:假手术组、模型组及温阳通脉方低剂量治疗组、中剂量治疗组和高剂量治疗组。将大鼠灌胃给药14天后,结扎心脏左冠状动脉前降支,按缺血30min再灌注120min的方法建立心肌缺血再灌注损伤模型。全自动生化分析仪检测各组大鼠的血清肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH);心电图检测大鼠ST段抬高情况;HE染色观察心脏组织病理形态学变化;免疫组织化学染色检测心肌组织中AQP1、AQP4蛋白的表达;Westernblot检测JAK2、p-JAK2、STAT3、p-STAT3、AQP1、AQP4的蛋白表达。结果与假手术组比较,模型组大鼠血清中的CK-MB、LDH含量显著上升(P<0.01),缺血30min与再灌注120min后心电图的ST段显著抬高(P<0.01),HE染色显示模型组的心肌细胞损伤严重,AQP1、AQP4免疫组织化学表达明显增多(P<0.01),JAK2、p-JAK2、STAT3、p-STAT3、AQP1、AQP4的蛋白表达均明显升高(P<0.01)。与模型组相比,给药大鼠CK-MB、LDH水平明显降低(P<0.01),缺血30min与再灌注120min后的ST段均降低(P<0.01),心肌细胞损伤均可见不同程度减轻,AQP1、AQP4免疫组织化学表达明显减少,JAK2、p-JAK2、STAT3、p-STAT3整体的表达呈升高趋势,尤其是p-JAK2和p-STAT3的表达(P<0.01),AQP1、AQP4的水平均有不同程度降低(P<0.01)。结论温阳通脉方的心肌保护作用可能与激活JAK2/STAT3信号通路,下调AQP1和AQP4的表达有关。

The Influence of Gastrodin on Expression of IL-10, STAT3 and JAK2 in Epileptic Rats’ Hippocampus

Objective: To explore the influence of gastrodin on IL-10, JAK2 and STAT3 in hippocampus of epileptic rats induced by pentylenetetrazol and the role of the IL-10 pathway in epilepsy. Methods: 50 adult male Wistar rats were randomly divided into 5 groups: normal control group (NC Group), epilepsy model group (EP Group), low doses of gastrodin + EP Group (GE1 Group), medium doses of gastrodin + EP Group (GE2 Group), high doses of gastrodin + EP Group (GE3 Group). EP group and GE Groups were injected subthreshold doses of pentylenetetrazole (PTZ) by intraperitoneal once a day until reaching the ignited standards. GE groups were respectively injected 4, 6, 8 mg/kg gastrodin by intraperitoneal. All groups were administered for 28 consecutive days. The behavioral changes of the rats were observed and recorded daily 1 hour after the injection. mRNAs of IL-10, STAT3 and JAK2 in hippocampus were measured by RT-qPCR, and proteins by Western blot. Results: Compared GE2 group with EP group, the incubation of seizure was significantly prolonged (P Conclusions: Gastrodin can increase the expression of IL-10, and reduce the expression of STAT3 and JAK2, which may play an antiepileptic effect through regulating JAK2/STAT3 signaling pathways by IL-10.

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.

miR-221-3p对肝癌细胞增殖的影响及相关机制研究

目的探讨微小RNA-221-3p(miR-221-3p)对肝癌细胞增殖的影响及其相关机制。方法采用生物学数据库分析肝癌中miR-221-3p的表达水平及其对患者预后的影响。CCK-8、EdU实验分析miR-221-3p对肝癌细胞增殖的影响。采用生物信息学及Western blot探讨miR-221-3p调控肝癌细胞增殖的相关机制。结果miR-221-3p在肝癌中呈高表达,其表达水平与患者总生存率无关。miR-221-3p沉默后肝癌细胞增殖能力减弱,而过表达miR-221-3p后肝癌细胞增殖能力增强。miR-221-3p可以通过负调控残留样蛋白质家族成员4(VGLL4)进而激活JAK2/STAT3信号通路。结论miR-221-3p促进HCC细胞的增殖,其机制可能与负调控VGLL4进而激活JAK2/STAT3信号通路有关。

TAK-242通过调控JAK2/STAT3通路抑制小鼠心肌缺血再灌注损伤炎症反应

目的:探讨Toll样受体4拮抗剂TAK-242抑制小鼠心肌缺血/再灌注损伤(ischemia/reperfusion,I/R)炎症反应的分子机制。方法:选用48只雄性C57BL/6小鼠随机分为4组:假手术组(sham)、模型组(I30min/R24h)、给药组[I/R+TAK-242(3 mg·kg-1)]、干预组[I/R+TAK-242+AG490(15 mg·kg-1)]。再灌注24 h后心脏超声检测小鼠心功能,氯化三苯基四氮唑(TTC)染色法测定心肌梗死面积,HE染色观察心肌病理改变,WB检测心肌JAK2/STAT3磷酸化水平,ELISA检测血清IL-6、TNF-α、IL-10和高迁移率族蛋白B1(HMGB1)浓度。结果:与sham组比较,I/R组小鼠左心室收缩期直径(LVIDs)延长(P<0.01),左心室射血分数(LVEF)和左心室短轴缩短分数(LVFS)显著降低(P<0.001或P<0.01),心梗面积明显增加并出现心肌炎性浸润,心肌p-JAK2/p-STAT3表达明显升高(P<0.01或P<0.05),血清IL-6、IL-10、TNF-α和HMGB1水平显著升高(P<0.001或P<0.01)。与I/R组比较,TAK-242给药组小鼠LVIDs缩短(P<0.05),LVEF和LVFS显著升高(P<0.01或P<0.05),心梗面积缩小(P<0.01),心肌炎症浸润减轻,心肌p-JAK2/p-STAT3表达降低(P<0.01或P<0.05),血清IL-6和TNF-α水平明显下降(P<0.001或P<0.01),而IL-10和HMGB1浓度进一步升高(P<0.01)。与TAK-242给药组比较,AG490干预可显著加强TAK-242治疗作用,包括心肌收缩功能增强,心梗面积缩小及炎性浸润程度减轻,心肌p-JAK2/p-STAT3表达降低(P<0.05),血清IL-6、TNF-α浓度下降而IL-10、HMGB1浓度升高(P<0.01或P<0.05)。结论:Toll样受体4拮抗剂TAK-242抑制小鼠I/R炎症反应与JAK2/STAT3信号通路失活有关。