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.

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.

免疫趋化因子CXCR4通过IL-6/STAT3信号通路调控胃癌炎症因子分泌和凋亡

目的研究趋化因子CXCR4通过IL-6/STAT3信号通路对胃癌细胞凋亡的影响及对相关机制。方法荧光定量PCR和免疫印迹法(Western blotting)检测人胃癌组织和癌旁组织(PCT)中免疫趋化因子CXCR4 mRNA和蛋白表达。其次,在人胃癌细胞系SGC7901中转染敲低和过表达CXCR4慢病毒。TUNEL染色检测SGC7901细胞凋亡情况;MTT检测SGC7901细胞增殖情况;Western blotting检测细胞凋亡相关蛋白Bax和Bcl2表达。进一步地,酶联免疫吸附试验(ELISA)检测炎症因子分泌水平;实时荧光定量PCR(RT-PCR)检测IL-6/STAT3信号通路IL-6 mRNA表达;Western blot检测STAT3-Ser727蛋白表达。此外,SGC7901细胞中敲低CXCR4后联合IL-6/STAT3信号通路激动剂脂多糖(LPS),同时SGC7901细胞中过表达CXCR4后联合IL-6/STAT3信号通路抑制剂angoline或bruceantinol,然后用TUNEL染色检测SGC7901细胞凋亡情况、Western blot检测细胞凋亡相关蛋白Bax和Bcl2表达。结果RT-PCR和Western blot检测发现,相比较于癌旁组织,胃癌组织中免疫趋化因子CXCR4表达升高。单细胞凝胶电泳检测表明,敲低和过表达CXCR4不会引起SGC7901细胞DNA损伤。TUNEL染色、MTT细胞增殖检测和Western blot检测发现,SGC7901细胞中敲低CXCR4促进细胞凋亡,过表达CXCR4抑制细胞凋亡。ELISA检测发现,SGC7901细胞中敲低CXCR4促进促炎因子IL-1β和TNF-α表达,抑制抑炎因子IL-10和TGF-β表达,而过表达CXCR4则表现出相反的效果。此外,SGC7901细胞中敲低CXCR4后激活IL-6/STAT3信号通路可降低SGC7901细胞凋亡,而抑制IL-6/STAT3信号通路可明显抑制过表达CXCR4对SGC7901细胞增殖的诱导作用。结论免疫趋化因子CXCR4通过IL-6/STAT3信号通路调控胃癌细胞凋亡和炎症因子分泌过程。

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.

IL-37b suppresses epithelial mesenchymal transition in hepatocellular carcinoma by inhibiting IL-6/STAT3 signaling

Background: Interleukin-37 b(IL-37 b), a vital negative regulator of the innate immune system, has been reported to be a tumor inhibitor in different type of cancers. However, little is known about the relationship between IL-37 b and hepatocellular carcinoma(HCC). The present study aimed to investigate the potential roles of IL-37 b in HCC progression. Methods: Subjects( n = 237) were recruited, and serum IL-37 b was measured using ELISA. The tumorsuppressive capacity and underlying mechanisms of IL-37 b in HCC were investigated in vitro and in vivo. Results: Compared to healthy controls, serum IL-37 b levels were elevated in chronic hepatitis B(CHB) patients but decreased significantly in HBV-HCC patients, especially for those with portal venous tumor thrombus. Low level serum IL-37 b in HBV-HCC patients correlated with high HCC stage and poor overall survival and disease-free survival. In vitro and in vivo, recombinant human IL-37 b inhibited proliferation and metastasis in HCC cells. Furthermore, IL-37 b inhibited epithelial mesenchymal transition in HCC cells in vitro by downregulating IL-6, pSTAT3(Y705), N-cadherin, and vimentin expression and by upregulating E-cadherin expression. These effects were partially reversed by transfection of adenovirus encoding human IL-6. Conclusions: IL-37 b inhibits HCC growth, metastasis and epithelial mesenchymal transition by regulating IL-6/STAT3 signaling. Serum IL-37 b may be a biomarker for HBV-HCC and its staging.

四逆散及其拆方对实验性溃疡性结肠炎大鼠IL-4/STAT6通路的影响

目的:从基因水平进一步探讨四逆散调控实验性溃疡性结肠炎大鼠IL-4/STAT6信号通路的作用机制。方法:将实验动物分为6组,采用免疫法造模,以大鼠IL-4、SOCS1、STAT6的基因表达作为观察与检测指标。结果:四逆散组实验大鼠IL-4、SOCS1表达显著增加(P<0.05),而STAT6显著降低(P<0.05);三个拆方组中,柴芍配伍对IL-4、SOCS1 mRNA表达的影响与四逆散无显著性差异(P>0.05),但STAT6 mRNA的表达显著高于正常组(P>0.05);与柴枳甘组、芍枳甘组比较,柴芍组IL-4、SOCS1 mRNA表达明显升高(P>0.05),STAT6 mRNA的表达显著降低(P>0.05)。结论:四逆散能够通过刺激IL-4、SOCS1 mRNA的表达,抑制STAT6mRNA的表达来调节IL-4/STAT6通路,干预实验性UC。方中柴芍配伍作用明显,主要体现在对IL-4、SOCS1mRNA表达的影响上;柴、芍分别配伍柴枳后,对TL-4/STAT6通路的调节作用明显降低。

沙眼衣原体持续感染对Hela细胞TLR4/IL-6/STAT3信号通路的影响

目的:分析沙眼衣原体(Chlamydia trachomatis,Ct)持续感染对靶细胞TLR4/IL-6/STAT3信号通路的影响。方法:利用Hela细胞分别建立Ct急性感染及持续性感染模型,通过qRT-PCR、ELISA等方法比较Ct感染过程中靶细胞TLR4、STAT3、IL-6转录水平及细胞因子IL-6分泌量的变化。结果:Ct感染后靶细胞TLR4、IL-6、STAT3转录水平及细胞因子IL-6分泌量均呈现时间相关性上调,且持续性感染状态下比急性感染状态下的上调更为显著;IL-6/STAT3的表达量与TLR4转录水平正相关。结论:Ct持续感染过程中TLR4的持续活化可大幅上调IL-6/STAT3信号通路表达,可能参与了Ct持续感染后慢性炎性损伤过程。

IL-4RA基因转染对哮喘模型气道STAT6的干预作用

目的:观察呼吸道转染IL-4RA基因对哮喘模型气道STAT6的干预作用。方法:BALB/c小鼠被随机分为五组,空白对照组、哮喘模型组、空载体干预组、激素治疗组、目的基因干预组。除空白对照组外其他各组小鼠在试验第1天和第7天和第14天通过腹腔注射0.2ml混有40mg氢氧化铝和10μg OVA的pH7.4 PBS致敏,空载体干预组和基因干预组在试验第12天连续给药三天,激素治疗组在激发的同时给予激素雾化吸入一周,在试验第15天用5%的OVA对小鼠进行雾化激发,连续一周。在最后一次激发后24小时放血杀死小鼠,检测血浆IgE水平和血中嗜酸细胞计数,留取肺组织标本做进一步的分析:肺组织用10%的甲醛固定,通过免疫组化的方法对STAT6的表达进行检测。结果:逆转录病毒载体携带目的基因IL-4RA成功地整合到了宿主基因组中,通过逆转录病毒转染的IL-4RA的基因高表达成功地阻止在哮喘模型鼠气道由IL-4和IL-13诱发的气道嗜酸细胞浸润,另外,逆转录病毒介导的IL-4RA在气道的表达明显地阻止了哮喘相关的气道STAT6水平,因此基因治疗可能会成为治疗慢性哮喘气道炎症和哮喘症状的极有潜力的药物。结论:IL-4RA基因转染阻止了哮喘模型气道STAT6的产生。

刘氏小儿推拿“推胸背法”对咳嗽变异性哮喘模型大鼠IL-4/STAT6信号通路的影响

目的:探讨刘氏小儿推拿"推胸背法"对咳嗽变异性哮喘模型大鼠IL-4/STAT6信号通路的影响。方法:采用卵清蛋白和氢氧化铝致敏及雾化激发的方法造模,分别采用孟鲁司特钠咀嚼片、"推胸背法"和推药结合法干预。采用WB法检测肺组织中p-STAT6的表达;采用ELISA法检测血清中IL-4含量和肺泡灌洗液中IL-4R含量。结果:模型组、西药组、推拿组和推药结合组的喷嚏、搔鼻次数与空白组比较,差异有统计学意义(P<0.01),说明造模成功。推拿组、西药组和推药结合组喷嚏、搔鼻次数比模型组明显减少(P<0.05);推拿组和西药组血清IL-4水平、肺泡灌洗液IL-4R水平和肺组织p-STAT6含量均低于模型组(P<0.05);推药结合组以上指标均低于推拿组和西药组(P<0.05)。结论:刘氏小儿推拿可能通过IL-4/STAT6信号通路调节咳嗽变异性哮喘大鼠的气道变应性炎症,同时能提高孟鲁司特钠咀嚼片的临床疗效。

IL-4、IL-12、IL-6/STAT/SOCS途径对Th细胞分化调节机制的研究进展

Th细胞分化为Th1和Th2细胞是受多种细胞因子调控的。已有研究资料表明 :IL 4、IL - 12和IL 6通过STAT/SOCS信号途径经对Th细胞的分化进行调节。IL 12 /STAT4 /SOCS3使CD4 + 和Th2向Th1分化和极化 ,IL 4 /STAT6 /SOCS1使Th0B向Th2分化。IL 6则通过STAT3可以上调CD4 + T细胞SOCS 1的表达 ,这是由于IL 6干扰IFN γ受体的信号转导 ,阻断了IFN γ对IFN γ基因的自我调节 ,因此实现了IL 6对Th1细胞的负性调控。据此认为IL 6启动CD4 + T细胞分化与抑制CD4 + T分化为Th1细胞是两种独立的分子机制 ,它在平衡Th1/Th2的免疫反应中起着重要的调节作用。

红外线局部照射联合皮肤再生医疗技术治疗开腹胃肠外科术后切口感染效果及对IL-4/IL-13/STAT6信号通路影响

背景切口感染是开腹胃肠外科术常见并发症及临床亟待解决的问题.红外线局部照射联合皮肤再生医疗技术能通过改善切口局部血液循环,降低致痛化学介质作用,抑制切口过度炎症反应,并通过加快渗出物吸收,缓解局部炎症.目的探讨红外线局部照射联合皮肤再生医疗技术治疗开腹胃肠外科术后切口感染效果及对白介素-4(interleukin-4,IL-4)/白介素-13(interleukin-13,IL-13)/转录激活子6(signal transducer and activator of transcription 6,STAT6)信号通路影响.方法按照1:1试验设计原则和电脑随机数字表法将2019-03/2021-06我院收治的66例开腹胃肠外科术后切口感染患者,随机分为对照组、实验组进行前瞻性研究,各33例.两组均给予抗菌药物治疗,在此基础上,对照组给予皮肤再生医疗技术,实验组给予红外线局部照射联合皮肤再生医疗技术.比较两组疗效、炎症指标[C反应蛋白(C-reactive protein,CRP)、降钙素原(procalcitonin,PCT)、白细胞(WBC)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)]、IL-4 mRNA、IL-13 mRNA、STAT6 mRNA、换药次数、红肿痛缓解时间、病原菌培养转阴时间、住院时间、温哥华瘢痕量表(vancouver scar scale,VSS)评分.结果两组疗效分布情况比较,差异有统计学意义,实验组显效率为75.76%,高于对照组的42.42%(P<0.05);实验组治疗3 d后、治疗5 d后CRP、PCT、WBC、TNF-α低于对照组(P<0.05);实验组治疗3 d后、治疗5 d后IL-4 mRNA、IL-13 mRNA、STAT6 mRNA低于对照组(P<0.05);实验组换药次数、红肿痛缓解时间、病原菌培养转阴时间、住院时间低于对照组(P<0.05);实验组VSS量表色泽、血管、柔软性、厚度、疼痛评分低于对照组(P<0.05).结论红外线局部照射联合皮肤再生医疗技术治疗开腹胃肠外科术后切口感染,能增强疗效,改善炎症反应与切口愈合质量,加快患者临床症状体征缓解和恢复进程.

五味子乙素对过敏性哮喘小鼠IL-4/IL-13/STAT6通路及气道杯状细胞凋亡的影响

目的:探究五味子乙素对过敏性哮喘小鼠的治疗作用及对IL-4/IL-13/STAT6信号通路的影响。方法:BALB/c小鼠随机分为6组:空白对照组、模型组、五味子乙素低(20 mg/kg)、中(30 mg/kg)、高(40 mg/kg)剂量组和地塞米松组(2 mg/kg),每组12只。除空白对照组外,其他各组均构建过敏性哮喘小鼠模型,造模同时分别灌胃给予相应药物。末次OVA激发24 h后,ELISA检测支气管肺泡灌洗液(BALF)中炎症因子IL-4、IL-13、IFN-γ水平。取出肺脏,计算肺/体质量指数。HE染色检测小鼠肺组织形态学变化,并进行炎症反应评分。TUNEL和PAS染色结合检测小鼠肺组织杯状细胞凋亡率;Western blot检测小鼠肺组织IL-4、IL-13、p-STAT6蛋白表达。结果:与空白对照组相比,模型组小鼠肺组织结构损伤严重,有大量炎症细胞浸润,肺/体质量指数、炎症反应评分、气道杯状细胞凋亡率、BALF中IL-4、IL-13水平及肺组织中IL-4、IL-13、p-STAT6蛋白表达显著升高(P<0.05),IFN-γ水平显著降低(P<0.05);与模型组相比,五味子乙素各剂量组和地塞米松组肺组织结构较完整,病变程度减轻,肺/体质量指数、炎症反应评分、气道杯状细胞凋亡率、BALF中IL-4、IL-13水平及肺组织中IL-4、IL-13、p-STAT6蛋白表达显著降低(P<0.05),IFN-γ水平显著升高(P<0.05),且呈剂量依赖关系。五味子乙素高剂量组和地塞米松组小鼠肺/体质量指数、炎症因子水平(IL-4、IL-13、IFN-γ)、细胞凋亡率及IL-4、IL-13、p-STAT6蛋白表达差异均无统计学意义(P>0.05)。结论:五味子乙素可能通过抑制IL-4/IL-13/STAT6信号通路减轻过敏性哮喘小鼠炎症反应,缓解肺组织病理损伤。

MiR-146a-5p targeting SMAD4 and TRAF6 inhibits adipogenensis through TGF-β and AKT/mTORC1 signal pathways in porcine intramuscular preadipocytes

Background: Intramuscular fat(IMF) content is a vital parameter for assessing pork quality. Increasing evidence has shown that microRNAs(miRNAs) play an important role in regulating porcine IMF deposition. Here, a novel miRNA implicated in porcine IMF adipogenesis was found, and its effect and regulatory mechanism were further explored with respect to intramuscular preadipocyte proliferation and differentiation.Results: By porcine adipose tissue miRNA sequencing analysis, we found that miR-146a-5p is a potential regulator of porcine IMF adipogenesis. Further studies showed that miR-146a-5p mimics inhibited porcine intramuscular preadipocyte proliferation and differentiation, while the miR-146a-5p inhibitor promoted cell proliferation and adipogenic differentiation. Mechanistically, miR-146a-5p suppressed cell proliferation by directly targeting SMAD family member 4(SMAD4) to attenuate TGF-β signaling. Moreover, miR-146a-5p inhibited the differentiation of intramuscular preadipocytes by targeting TNF receptor-associated factor 6(TRAF6) to weaken the AKT/mTORC1 signaling downstream of the TRAF6 pathway.Conclusions: MiR-146a-5p targets SMAD4 and TRAF6 to inhibit porcine intramuscular adipogenesis by attenuating TGF-β and AKT/mTORC1 signaling, respectively. These findings provide a novel miRNA biomarker for regulating intramuscular adipogenesis to promote pork quality.

布地奈德通过调节IL-4/IL-13/STAT6影响大鼠黏膜重塑

目的探讨布地奈德通过IL-4/IL-13/STAT6信号通路对变应性鼻炎(allergic rhinitis,AR)大鼠鼻黏膜重塑的影响。方法 102只大鼠采用随机数字表法分为3组:空白组、AR-PBS组、AR-布地奈德组,每组34只。AR-PBS组及AR-布地奈德组大鼠为建模组大鼠,均采用卵清蛋白致敏的方法构建AR模型。建模成功后,使用微量移液枪双侧滴鼻治疗。AR-PBS组给予(PBS)治疗,AR-布地奈德组给予布地奈德(倍受您)治疗,空白组大鼠给予等量PBS治疗,治疗时间持续7 d。采用动物评价量表对各组大鼠行为学进行评分;ELISA法测定血清组胺(His)及Ig E;液相芯片法测定血清及肺泡灌洗液IL-4、IL-13浓度;定量PCR检测STAT6基因表达,Western blot检测鼻黏膜总STAT6及磷酸化p-STAT6蛋白表达;鼻中隔黏膜HE染色及透射电镜扫描观察鼻黏膜重塑情况;采用SPSS 23.0统计软件对数据进行分析处理。结果建模组大鼠建模结束时动物行为学评分均>5分,血清His、Ig E,血清及肺泡灌洗液IL-4及IL-13浓度以及鼻中隔黏膜STAT6及p-STAT6表达均显著高于空白组(P<0.01)。建模组大鼠采用不同方法治疗,AR-布地奈德组干预7 d后,血清His、Ig E,血清及肺泡灌洗液IL-4、IL-13浓度,以及鼻中隔黏膜STAT6、p-STAT6表达均下调,显著低于AR-PBS组,差异有统计学意义(P<0.01)。HE染色及透射电镜结果显示建模组大鼠建模结束时鼻纤毛减少,布地奈德治疗后纤毛具有一定程度的恢复,治疗前后细胞连接无显著差异。结论布地奈德通过降低IL-4、IL-13、p-STAT6从而改善AR大鼠鼻黏膜症状。

Ramulus Cinnamomi extract attenuates neuroinflammatory responses via downregulating TLR4/MyD88 signaling pathway in BV2 cells

Ramulus Cinnamomi （RC）, a traditional Chinese herb, has been used to attenuate inflammatory responses. The purpose of this study was to investigate the effect of RC extract on lipopolysaccharide （LPS）-induced neuroinflammation in BV2 microglial cells and the underlying mechanisms involved. BV2 cells were incubated with normal medium （control group）, LPS, LPS plus 30 pg/mL RC extract, or LPS plus 100 pg/mL RC extract. The BV2 cell morphology was observed under an optical microscope and cell viability was detected by MTT assay. Nitric oxide level in BV2 cells was detected using Griess regents, and the levels of interleukin-6, interleukin-1 β, and tumor necrosis factor u in BV2 cells were determined by ELISA. The expression levels of cyclooxygenase-2, Toll-like receptor 4 and myeloid differentiation factor 88 proteins were detected by western blot assay. Compared with the LPS group, both 30 and 100 μg/mL RC extract had no significant effect on the viability of BV2 cells. The levels of nitric oxide, interleukin-6, interleukin-1β and tumor necrosis factor ct in BV2 cells were all significantly increased after LPS induction, and the levels were significantly reversed after treatment with 30 and 100 μg/mL RC extract. Furthermore, RC extract significantly inhibited the protein expression levels of cyclooxygenase-2, Toll-like receptor 4 and myeloid differentiation factor 88 in LPS-induced BV2 cells. Our findings suggest that RC extract alleviates neuroinflammation by downregulating the TLR4/MyD88 signaling pathway.

KIAA1199 induces advanced biological behavior and development of ovarian cancer through activation of the IL-6/STAT3 pathway

Recently,abnormal expression of KIAA1199 has been detected in Epithelial Ovarian Cancer(EOC).However,the underlined anti-ovarian cancer mechanism of KIAA1199 remains to be enlightened.In our study,we performed to elucidate the effects of KIAA1199 on the advanced biological behavior of EOC cells through activation of the IL-6/STAT3 pathway.Confirmed by immunohistochemistry,KIAA1199 was highly expressed in ovarian borderline and malignant epithelial tumors.A retrospective analysis found that EOC patients with low expression of KIAA1199 had a significantly higher 5-year survival rate than those with high expression.Mechanistically,IL-6 was used to stimulate EOC cells,and the expression of KIAA1199,STAT3 and p-STAT3 increased after IL-6 stimulation.These results could show that KIAA1199 is transcriptionally activated by IL6/STAT3 pathway,thereby accelerating the deterioration of EOC.KIAA1199 could also be used as a poor prognosis factor and potential target in treatment.

YD383、YD439对JAK-STAT6信号传导通路作用的特异性研究

目的:探讨化合物YD383和YD439对STAT6信号传导通路的作用是否具有特异性。方法:通过瞬时转染的方法将分别依赖于STAT1和STAT6活性的带有荧光素酶报告基因的质粒转染至HeLa细胞株,两种细胞株在各自相应的细胞因子(IFN-γ或IL-4)刺激下激活JAK-STAT1或JAK-STAT6信号传导通路。通过测定荧光素酶的活性来评价细胞株在化合物存在的情况下报告基因的表达。结果:YD383和YD439均能降低相应细胞因子刺激引起的报告基因的表达,并且具有剂量依赖性。随着化合物剂量的增加,报告基因的表达降低(P<0.05)。但化合物引起的两种转染细胞报告基因表达降低的差异比较无统计学意义(P>0.05)。结论:两种化合物对JAK-STAT1和JAK-STAT6信号传导通路均有抑制作用,对JAK-STAT6通路的抑制不具有特异性。

INHIBITION OF IL-6-INDUCED STAT3 ACTIVATION IN MYELOMA CELLS BY PROTEIN KINASE A

Objective: To investigate the regulation effect of protein kinase A on IL-6-induced STAT3 activation in myeloma cells. Methods: Two human myeloma cell lines-Sko-007 and U266 were pretreated with Forskolin, a protein kinase A antagonist, and then stimulated by IL-6. The activation state of STAT3 in these two cells were examined by electrophoretic mobility shift assay (EMSA). Results: Although PKA pathway itself doesn’t participate in IL-6 signal transduction in Sko-007 and U266 cells, activation of protein kinase A can inhibit IL-6-induced STAT3 activation in these two cell lines. Conclusion: There exists an inhibitory effect of protein kinase A on STAT3 activation in human myeloma cells treated by IL-6.