Role of Toll-like receptor 4 and Janus kinase and signal transducer and activator of transcription signal transduction pathway in sepsis-induced brain damage

The Janus kinase and signal transducer and activator of transcription （JAK/STAT） signal transduction pathway is involved in sepsis-induced functional damage to the heart, liver, kidney, and other organs. However, the cellular and molecular mechanisms underlying sepsis-induced brain damage remain elusive. In the present study, we found severe loss of neurons in the hippocampal CA1 region in rats with sepsis-induced brain damage following intraperitoneal injection of endotoxin, The expression of toll-like receptor 4, tumor necrosis factor a, and interleukin-6 was significantly increased in brain tissues following lipopolysaccharide exposure. AG490 （JAK2 antagonist） and rapamycin （STAT3 antagonist） significantly reduced neuronal loss and suppressed the increased expression of toll-like receptor 4, tumor necrosis factor a, and interleukin-6 in the hippocampal CA1 region in sepsis-induced brain damaged rats. Overall, these data suggest that blockade of the JAK/STAT signal transduction pathway is neuroprotective in sepsis-induced brain damage via the inhibition of toll-like receptor 4, tumor necrosis factor a, and interleukin-6 exoression.

Mechanism of ELL-associated factor 2 and vasohibin 1 regulating invasion,migration,and angiogenesis in colorectal cancer

BACKGROUND As a novel endogenous anti-angiogenic molecule, vasohibin 1(VASH1) is not only expressed in tumor stroma, but also in tumor tissue. Moreover, studies have shown that VASH1 may be a prognostic marker in colorectal cancer(CRC). Knockdown of VASH1 enhanced transforming growth factor-β1(TGF-β1)/Smad3 pathway activity and type Ⅰ/Ⅲ collagen production. Our previous findings suggest that ELL-associated factor 2(EAF2) may play a tumor suppressor and protective role in the development and progression of CRC by regulating signal transducer and activator of transcription 3(STAT3)/TGF-β1 signaling pathway. However, the functional role and mechanism of VASH1-mediated TGF-β1 related pathway in CRC has not been elucidated.AIM To investigate the expression of VASH1 in CRC and its correlation with the expression of EAF2. Furthermore, we studied the functional role and mechanism of VASH1 involved in the regulation and protection of EAF2 in CRC cells in vitro.METHODS We collected colorectal adenocarcinoma and corresponding adjacent tissues to investigate the clinical expression of EAF2 protein and VASH1 protein in patients with advanced CRC. Following, we investigated the effect and mechanism of EAF2 and VASH1 on the invasion, migration and angiogenesis of CRC cells in vitro using plasmid transfection.RESULTS Our findings indicated that EAF2 was down-regulated and VASH1 was upregulated in advanced CRC tissue compared to normal colorectal tissue. KaplanMeier survival analysis showed that the higher EAF2 Level group and the lower VASH1 Level group had a higher survival rate. Overexpression of EAF2 might inhibit the activity of STAT3/TGF-β1 pathway by up-regulating the expression of VASH1, and then weaken the invasion, migration and angiogenesis of CRC cells.CONCLUSION This study suggests that EAF2 and VASH1 may serve as new diagnostic and prognostic markers for CRC, and provide a clinical basis for exploring new biomarkers for CRC. This study complements the mechanism of EAF2 in CRC cells, enriches the role and mechanism of CRC cellderived VASH1, and provides a new possible subtype of CRC as a therapeutic target of STAT3/TGF-β1 pathway.

Regulation and function of signal transducer and activator of transcription 3

Signal transducer and activator of transcription 3(STAT3), a member of the STAT family, is a key regulator of many physiological and pathological processes. Significant progress has been made in understanding the transcriptional control, posttranslational modification, cellular localization and functional regulation of STAT3. STAT3 can translocate into the nucleus and bind to specific promoter sequences, thereby exerting transcriptional regulation. Recent studies have shown that STAT3 can also translocate into mitochondria, participating in aerobic respiration and apoptosis. In addition, STAT3 plays an important role in inflammation and tumorigenesis by regulating cell proliferation, differentiation and metabolism. Conditional knockout mouse models make it possible to study the physiological function of STAT3 in specific tissues and organs. This review summarizes the latest advances in the understanding of the expression, regulation and function of STAT3 in physiological and tumorigenic processes.

Silencing of signal transducer and activator of transcription 3 expression by RNA interference suppresses growth of human hepatocellular carcinoma in tumor-bearing nude mice

AIM:To explore the effect of silencing of signal transducer and activator of transcription 3(STAT3) expression by RNA interference(RNAi) on growth of human hepatocellular carcinoma(HCC) in tumor-bearing nude mice in vivo.METHODS:To construct the recombinant plasmid of pSilencer 3.0-H1-STAT3-siRNA-GFP(pSH1-siRNA-STAT3) and establish the tumor-bearing nude mouse model of the HCC cell line SMMC7721,we used intratumoral injection together with electroblotting to transfect the recombinant plasmid pSH1-siRNA-STAT3 into the transplanted tumor.The weight of the nude mice and tumor volumes were recorded.STAT3 gene transcription was detected by semi-quantitative reverse transcription polymerase chain reaction(RTPCR).Level of protein expression and location of STAT3 were determined by Western blotting and immunohistochemical staining.STAT3-related genes such as survivin,c-myc,VEGF,p53 and caspase3 mRNA and protein expression were detected in tumor tissues at the same time.The terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL) assay was used to detect apoptosis of tumor cells.RESULTS:The weight of the treated nude mice increased,and the tumor volume decreased markedly compared with those of the mock-treated and negative control groups(P < 0.01).The results of RT-PCR and Western blotting showed that mRNA and protein levels of STAT3 declined markedly in the treated group.The change in STAT3-related gene expression in tumor tissues at the mRNA and protein level also varied,the expression of survivin,VEGF and c-myc were obviously reduced,and expression of p53 and caspase3 increased(P < 0.01).Most of the tumor tissue cells in the treated group developed apoptosis that was detected by TUNEL assay.CONCLUSION:Silencing of STAT3 expression by RNAi significantly inhibits expression of STAT3 mRNA and protein,and suppresses growth of human HCC in tumor-bearing nude mice.The mechanism may be related to down-regulation of survivin,VEGF and c-myc and up-regulation of p53 and caspase3 expression.Accordingly,the STAT3 gene may act as an important and effective target in gene therapy of HCC.

Expression of p-STAT3 and vascular endothelial growth factor in MNNG-induced precancerous lesions and gastric tumors in rats

AIM: To investigate the dynamic expression of p-signal transducer and activator of transcription 3(STAT3) and vascular endothelial growth factor(VEGF) in the formation of gastric tumors induced by drinking water containing N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) in Wistar rats.METHODS: One hundred and twenty Wistar rats were randomly divided into two groups(60 in each group): Control group and Model group. The rats in each group were then randomly divided into three groups(20 in each group): C/M15, C/M25 and C/M40(15, 25 and 40 represent the number of feeding weeks from termination). Rats in the control group received normal drinking water and rats in the model group received drinking water containing 100 μg/m L MNNG. Stomach tissues were collected at the end of the 15 th, 25 th and 40 th week, respectively, for microscopic measurement using hematoxylin and eosin staining. The expression of p-STAT3 and VEGF in different pathological types of gastric tissue, including normal, inflammation, atrophy, hyperplasia and gastric stromal tumor, was observed by immunohistochemistry and Western blot, and the corelation between p-STAT3 and VEGF was analyzed. RESULTS:(1) The expression of p-STAT3 in tissue with gastritis, atrophy, dysplasia and gastric stromal tumor were significantly increased in the model group compared with the control group(2.5 ± 1.0, 2.75 ±0.36, 6.2 ± 0.45, 5.67 ± 0.55 vs 0.75 ± 0.36, P = 0.026, 0.035, 0.001, 0.002, respectively); the expression of p-STAT3 in tissue with dysplasia was higher than that in samples with gastritis or atrophy(6.2 ± 0.45 vs 2.5 ± 1.0, P = 0.006; 6.2 ± 0.45 vs 2.75 ± 0.36, P = 0.005, respectively); however, the expression of p-STAT3 in gastritis and atrophy was not significantly different(P > 0.05);(2) the expression of VEGF in tissue with gastritis, atrophy, dysplasia and gastric stromal tumor was significantly increased in the model group compared with normal gastric mucosa; and the expression of VEGF in tissue with dysplasia was higher than that in tissue with inflammation and atrophy(10.8 ± 1.96 vs 7.62 ± 0.25, P = 0.029; 10.8 ± 1.96 vs 6.26 ± 0.76, P = 0.033, respectively); similarly, the expression of VEGF in tissue with gastritis and atrophy was not significantly different(P > 0.05); and(3) the expression of VEGF was positively correlated with p-STAT3. CONCLUSION: p-STAT3 plays an important role in gastric cancer formation by regulating the expression of VEGF to promote the progression of gastric tumor from gastritis.

化瘀止痛贴膏调节IL-6/STAT3信号通路对急性软组织损伤大鼠炎症反应的影响

目的探讨化瘀止痛贴膏调节白细胞介素-6(IL-6)/信号转导与转录激活子3(STAT3)信号通路对急性软组织损伤(ASTI)大鼠炎症反应的影响。方法将SPF级SD大鼠随机分为对照组、模型组、化瘀止痛贴膏低、中、高剂量组、精制狗皮膏组。利用机械冲击挫伤法建立ASTI动物模型,建模成功后,各组贴敷相应药物,1次/d,每次给药8 h,连续7 d。对大鼠进行ASTI损伤评分;采用血液流变检测仪检测血液流变学指标;苏木精和伊红(HE)染色法观察大鼠损伤部位肌肉组织病理变化;酶联免疫吸附法(ELISA)检测各组大鼠损伤部位肌肉组织中IL-6、IL-β、肿瘤坏死因子-α(TNF-α)水平;实时荧光定量PCR(qRT-PCR)法检测损伤部位肌肉组织中IL-6与STAT3 mRNA表达;蛋白免疫印迹(Western blot)法检测大鼠损伤部位肌肉组织中IL-6/STAT3信号通路蛋白表达。结果与对照组比较,模型组大鼠软组织损伤评分、全血黏度、血浆黏度、红细胞压积、肌肉组织病理学评分、IL-6、IL-β、TNF-α水平、IL-6与STAT3 mRNA表达水平及IL-6、p-STAT3/STAT3蛋白表达水平升高(均P<0.05);与模型组比较,化瘀止痛贴膏低、中、高剂量组和狗皮膏药组大鼠软组织损伤评分、全血黏度、血浆黏度、红细胞压积、肌肉组织病理学评分、IL-6、IL-β、TNF-α水平、IL-6与STAT3 mRNA表达水平及IL-6、p-STAT3/STAT3蛋白表达水平降低(均P<0.05);化瘀止痛贴膏高剂量组与精制狗皮膏组大鼠以上指标差异均无统计学意义(P>0.05)。结论化瘀止痛贴膏可能通过下调IL-6/STAT3信号通路缓解ASTI大鼠的炎症反应。

子宫内膜癌组织NF-κB、STAT3蛋白对子宫内膜癌的诊断及预后价值意义分析

目的探讨与分析子宫内膜癌(EC)组织核因子κB(NF-κB)、信号转导与转录激活因子3(STAT3)蛋白对子宫内膜癌的诊断及预后价值意义。方法选择2019年9月—2022年11月石河子大学第一附属医院收治的88例子宫内膜癌患者作为研究对象,取所有患者术中切除的新鲜子宫内膜癌肿瘤组织标本(肿瘤组)和癌旁正常子宫内膜组织(癌旁组),采用免疫组化法检测NF-κB、STAT3蛋白表达阳性率,调查患者的病理特征、随访预后并进行相关性分析。结果肿瘤组NF-κB、STAT3蛋白表达阳性率显著高于癌旁组,差异具有统计学意义(P<0.05)。在88例患者中,不同年龄、临床分期、分化程度、肌层浸润、淋巴结转移患者的NF-κB、STAT3蛋白表达阳性率比较,差异具有统计学意义(P<0.05)。所有患者随访到2023年6月1日,生存患者NF-κB、STAT3蛋白表达阳性率明显低于死亡患者,差异具有统计学意义(P<0.05)。Cox回归分析显示NF-κB、STAT3蛋白表达阳性率为影响预后中位生存时间的重要因素,差异有统计学意义(P<0.05)。结论子宫内膜癌组织多伴随有NF-κB、STAT3蛋白的高表达,其表达水平与患者的临床病理特征和预后生存情况都存在相关性。

基于IL-6/STAT3信号通路探讨miR-34a-5p过表达对溃疡性结肠炎的影响及小檗碱的干预作用

目的基于白细胞介素6(IL-6)/信号转导和转录激活因子3(STAT3)信号通路探讨微小RNA-34a-5p(miR-34a-5p)过表达对溃疡性结肠炎(UC)的影响及小檗碱(BBR)的干预作用。方法体外传代培养人结肠癌HT-29细胞。取传3代、对数生长期、生长状态良好的HT-29细胞,随机分为空白对照组、NC mimics组、miR-34a-5p mimics组,NC mimics组和miR-34a-5p mimics组分别转染NC mimics、miR-34a-5p mimics,空白对照组不予转染。转染6 h更换新鲜培养基继续培养48 h,收集细胞,采用RT-qPCR法验证转染效率。收集三组转染后细胞,采用CCK-8法检测细胞活力;分离三组培养上清液,采用ELISA法检测IL-6、IL-1β、TNF-α含量;收集三组转染后细胞,分别采用RT-qPCR法、Western blotting法检测IL-6、STAT3 mRNA和蛋白表达。取HT-29细胞,加入LPS刺激48 h,建立UC细胞模型。取UC细胞,分别加入0、2.5、5、10、20、40、80、160、320μg/mL BBR干预24 h,采用CCK-8法检测细胞活力。随机将UC细胞分为模型组和BBR组,BBR组予BBR干预24 h,模型组不予BBR干预。分离两组培养上清液,采用ELISA法检测IL-6、IL-1β、TNF-α含量;取两组干预24 h细胞,采用RT-qPCR法检测miR-34a-5p以及IL-6、STAT3mRNA表达。结果miR-34a-5p mimics组miR-34a-5p相对表达量高于NC mimics组和空白对照组(P均<0.05),NC mimics组与空白对照组比较差异无统计学意义(P>0.05)。miR-34a-5p mimics组细胞活力低于NC mimics组和空白对照组(P均<0.05),NC mimics组与空白对照组比较差异无统计学意义(P>0.05)。miR-34a-5p mimics组培养上清液IL-6、IL-1β、TNF-α含量均低于NC mimics组和空白对照组(P均<0.05),NC mimics组与空白对照组比较差异均无统计学意义(P均>0.05)。miR-34a-5p mimics组IL-6、STAT3 mRNA以及IL-6蛋白相对表达量均低于NC mimics组和空白对照组(P均<0.05),NC mimics组与空白对照组比较差异均无统计学意义(P均>0.05)。40、80、160、320μg/mL BBR干预24 h UC细胞活力均高于0、2.5、5、10、20μg/mL BBR干预24 h UC细胞(P均<0.05),故选择20μg/mL BBR进行后续实验。BBR组培养上清液IL-6、IL-1β、TNF-α含量均低于模型组(P均<0.05);BBR组miR-34a-5p相对表达量高于模型组,IL-6、STAT3 mRNA相对表达量均低于模型组(P均<0.05)。结论miR-34a-5p过表达能够通过抑制IL-6/STAT3信号通路减轻UC炎症反应;BBR则能通过上调miR-34a-5p表达和抑制IL-6/STAT3信号通路,减轻UC炎症反应,阻延UC癌变。

(-)-Epigallocatechin-3-gallate inhibits growth of gastric cancer by reducing VEGF production and angiogenesis

AIM: To investigate the effect of (-)-epigallocatechin- 3-gallate (EGCG) on growth of gastric cancer and its possible mechanism. METHODS: Heterotopic tumors were induced by subcutaneously injection of SGC-7901 cells in nude mice. Tumor growth was measured by calipers in two dimensions. Tumor angiogenesis was determined with tumor microvessel density (MVD) by immunohistology. Vascular endothelial growth factor (VEGF) protein level and activation of signal transducer and activator of transcription 3 (Stat3) were examined by Western blotting. VEGF mRNA expression was determined by RT-PCR and VEGF release in tumor culture medium by ELISA. VEGF-induced cell proliferation was studied by MTT assay, cell migration by gelatin modified Boyden chamber (Transwell) and in vitro angiogenesis by endothelial tube formation in Matrigel. RESULTS: Intraperitoneal injection of EGCG inhibited the growth of gastric cancer by 60.4%. MVD in tumor tissues treated with EGCG was markedly reduced. EGCG treatment reduced VEGF protein level in vitro and in vivo. Secretion and mRNA expression of VEGF in tumor cells were also suppressed by EGCG in a dose-dependent manner. This inhibitory effect was associated with reduced activation of Stat3, but EGCG treatment did not change the total Stat3 expression. EGCG also inhibited VEGF-induced endothelial cell proliferation, migration and tube formation. CONCLUSION: EGCG inhibits the growth of gastriccancer by reducing VEGF production and angiogenesis, and is a promising candidate for anti-angiogenic treatment of gastric cancer.

子宫肌瘤组织中HMGA2、STAT3蛋白水平评估患者术后肌瘤复发价值

目的:探索子宫肌瘤患者肌瘤组织中高迁移率组蛋白A2(HMGA2)和信号转导与转录激活因子3(STAT3)蛋白水平与患者术后复发关系及评估价值。方法:选取2019年5月-2020年12月在本院接受腹腔镜子宫肌瘤剔除术治疗的子宫肌瘤患者168例,3年术后随访分为复发组(48例)及未复发组(120例)。收集患者一般资料,采用酶联免疫吸附实验检测患者术前血清孕酮(P)、黄体生成素(LH)、雌二醇(E_(2))、卵泡刺激素(FSH)及泌乳素(PRL)的水平,采用Western blot定量分析切除肌瘤组织中HMGA2、STAT3水平;受试者工作特征(ROC)曲线分析HMGA2、STAT3水平对子宫肌瘤患者术后复发的预测价值,多因素logistic回归分析其是否影响术后复发。结果:复发组血清P、LH、E_(2)、FSH、PRL水平高于未复发组,切除的肌瘤组织中HMGA2(1.42±0.28)、STAT3(1.33±0.24)水平均高于未复发组(1.03±0.21、1.02±0.18)(均P<0.05);ROC曲线分析显示,HMGA2联合STAT3评估患者术后肌瘤复发的曲线下面积(0.934)大于HMGA2(0.885)、STAT3(0.858)(均P<0.05)。多因素分析显示,术前血清P、FSH、PRL,以及肌瘤组织HMGA2、STAT3水平升高是子宫肌瘤患者术后复发的影响因素(均P<0.05)。结论:子宫肌瘤组织HMGA2、STAT3水平上调与术后复发有关,二者联合检测对辅助临床评估患者预后有较好价值。

连翘脂素调节JAK2/STAT3信号通路对前列腺癌增殖、凋亡和上皮间质转化的影响

目的探讨连翘脂素(PHI)对前列腺癌(PCa)增殖、凋亡和上皮间质转化的影响及其机制。方法体外培养DU-145和LNCap-FGC细胞,将细胞分为对照组(Control组)、连翘脂素组(PHI组)、Janus激酶2(JAK2)抑制剂组(Ag490组)、连翘脂素+激活剂组(PHI+Colivelin组),分别检测细胞活力、细胞克隆能力、划痕愈合率及细胞侵袭;免疫组化法分析E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)和波形蛋白(Vimentin)表达;Westernblot检测JAK2、p-JAK2、信号转导和转录激活因子3(STAT3)、p-STAT3蛋白表达。结果与Control组比较,PHI组与Ag490组DU-145和LNCap-FGC胞存活率、细胞克隆数、划痕愈合率、细胞侵袭数、N-cadherin、Vimentin及p-JAK2、p-STAT3表达显著降低,细胞凋亡率和E-cadherin表达显著升高(P<0.05);PHI组与Ag490组比较,差异无统计学意义(P>0.05)。与PHI组比较,PHI+Colivelin组细胞存活率、细胞克隆数、细胞划痕愈合、细胞侵袭数、N-cadherin、Vimentin及p-JAK2、p-STAT3表达显著增加,细胞凋亡率和E-cadherin显著降低(P<0.05)。结论PHI可抑制JAK2/STAT3信号通路抑制PCa细胞增殖并诱导细胞凋亡,逆转DU-145和LNCap-FGC上皮间质转化进程。

Induction of apoptosis by TPA and VP-16 is through translocation of TR3

AIM: To investigate the role of TR3 in induction of apoptosisin gastric cancer cells.METHODS: Human gastric cancer cell line, MGC80-3, wasused. Expression of TR3 mRNA and its protein was detectedby Northern blot and Westem blot. Localization of TR3protein was showed by immunofluorescence analysis underlaser-scanning confocal microscope. Apoptotic morphologywas observed by DAPI fluorescence staining, and apoptoticindex was counted among 1000 cells randomly. Stabletransfection assay was carried out by Lipofectamine.RESULTS: Treatment of MGC80-3 cells with TPA and VP-16resulted in apoptosis, accompanied by the repression ofBcl-2 protein in a time-dependent manner. At the sametime, TPA and VP-16 also up-regulated expression level ofTR3 mRNA in MGC80-3 cells that expressed TR3 mRNA.When antisense-TR3 expression vector was transfected intothe cells, expression of TR3 protein was repressed. In thiscase, TPA and VP-16 did not induce apoptosis. In addition,TPA and VP-16-induced apoptosis involved in translocationof TR3. In MGC80-3 cells, TR3 localized concentrative innucleus, after treatment of cells with TPA and VP-16, TR3translocated from nucleus to cytosol obviously. However,when this nuclear translocation was blocked by LMB,apoptosis was not occurred in MGC80-3 cells even in thepresence of TPA and VP-16.CONCLUSION: Induction of apoptosis by TPA and VP-16 isthrough induction of TR3 expression and translocation of TR3from nucleus to cytosol, which may be a novel signalpathway for TR3, and represent the new biological function ofTR3 to exert its effect on apoptosis in gastric cancer cells.

Regulation of the EGFR pathway by visfatin and its effect on cardiac hypertrophy

Objective:To investigate the regulation of the epidermal growth factor receptor(EGFR)pathway by visfatin and its effect on cardiac hypertrophy.Methods:60 Wistar male rats were randomly divided into control group,visfatin group and visfatin+AG1478 group,with 20 rats in each group.The cardiac mass index,left ventricular mass index and cardiomyocyte volume of rats in each group were calculated.The total protein content of each group of cardiomyocytes was detected by coomassie bright blue staining,and the protein expression was detected by Western blotting.Results:Compared with the control group,the cardiac mass index,left ventricular mass index,cardiomyocyte volume,protein content,and relative expressions of ANP and BNP were significantly increased in the visfatin group(P<0.05).The relative expression levels of EGFR,p-AKT,p-ERK1/2,p-STAT3,ANP and BNP in cardiac myocytes in the visfatin group were significantly higher than those in the control group and the visfatin+AG1478 group(P<0.05).Conclusion:Visfatin induces hypertrophy in cardiomyocytes by activating the EGFR signaling pathway.

芒柄花素调节JAK2/STAT3信号通路对妊娠高血压大鼠的治疗作用

目的:探讨芒柄花素(FMN)对妊娠高血压(PIH)大鼠的治疗作用及可能机制。方法:将妊娠大鼠分为正常组(Normal组,生理盐水)、PIH组(生理盐水)、FMN组[40 mg/(kg·d)FMN]、Janus激酶2(JAK2)抑制剂组[AG490组,5 mg/(kg·d)AG490]、FMN+JAK2抑制剂组[FMN+AG490组,40 mg/(kg·d)FMN+5 mg/(kg·d)AG490],每组12只。除Normal组外,其余各组大鼠连续4 d皮下注射125 mg/(kg·d)L-精氨酸甲酯构建PIH模型。观察大鼠尾静脉压、24 h尿蛋白含量,血清血管内皮因子[内皮素1(ET-1)、一氧化氮(NO)]、炎性因子[肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)]、心肌损伤指标[肌酸激酶同工酶MB(CK-MB)、心肌肌钙蛋白I(cTnI)]、肾损伤指标[血尿素氮(BUN)、血肌酐(SCr)]水平,心肌、肾组织病理学变化及心肌、肾组织中氧化应激指标[丙二醛(MDA)和总超氧化物歧化酶(T-SOD)]和JAK2/信号转导和转录激活因子3(STAT3)信号通路相关蛋白(JAK2、p-JAK2、STAT3、p-STAT3)表达。结果:给药结束后,与Normal组比较,PIH组大鼠尾静脉压、24 h尿蛋白含量升高,血清ET-1、TNF-α、IL-6、CK-MB、cTnI、BUN、SCr水平及心肌、肾组织中MDA含量和p-JAK2/JAK2、p-STAT3/STAT3比值升高(P<0.05),血清NO水平和心肌、肾组织中T-SOD活性降低(P<0.05),且心肌、肾组织存在明显的病理损伤。与PIH组比较,FMN组、AG490组和FMN+AG490组大鼠尾静脉压、24 h尿蛋白含量降低,血清ET-1、TNF-α、IL-6、CK-MB、cTnI、BUN、SCr水平及心肌、肾组织中MDA含量和p-JAK2/JAK2、p-STAT3/STAT3比值降低(P<0.05),血清NO水平和心肌、肾组织中T-SOD活性升高(P<0.05),且心肌、肾组织病理损伤均有所改善;其中FMN+AG490组上述指标变化均显著优于FMN组、AG490组。结论:FMN可能通过抑制JAK2/STAT3信号通路,减轻心肌、肾组织中氧化应激和全身炎症反应,改善血管内皮舒缩功能,降低血压,减轻PIH大鼠心肌、肾组织损伤。

Overexpression of ELL-associated factor 2 suppresses invasion,migration,and angiogenesis in colorectal cancer

BACKGROUND The androgen responsive gene,ELL-associated factor 2(EAF2),expressed in benign prostate tissues,has been shown to play an important role in tumor suppression in a variety of malignant tumors.In addition,some scholars found that EAF2 frameshift mutations are associated with intratumor heterogeneity in colorectal cancer(CRC)and inactivation of EAF2 in microsatellite instability-high CRC.However,the molecular mechanism by which EAF2 is involved in CRC invasion and metastasis remains unclear.AIM To determine the clinical value of expression of EAF2 protein in CRC,and to study the effects of EAF2 on the invasion,migration,and angiogenesis of CRC cells in vitro.METHODS In this study,we collected colorectal adenocarcinoma and corresponding adjacent tissues to investigate the clinical expression of EAF2 protein in patients with advanced CRC.Subsequently,we investigated the effect of EAF2 on the invasion,migration,and angiogenesis of CRC cells in vitro using plasmid transfection.RESULTS EAF2 protein was lowly expressed in cancer tissues of patients with advanced CRC.Kaplan-Meier survival analysis showed that the survival rate of the high EAF2 level group was higher than that of the low EAF2 level group.CONCLUSION Our results demonstrated that EAF2,as a tumor suppressor,may inhibit the invasion,metastasis,and angiogenesis of CRC cells by regulating the signal transducer and activator of transcription 3/transforming growth factor-β1 crosstalk pathway,and play a cancer suppressive and protective role in the occurrence and development of CRC.Our findings are of great significance to provide a new idea and theoretical basis for the targeted diagnosis and treatment of CRC.

Effects of astragalus injection on the skin lesion degree and Caspase-14,SOCS1 and STAT3 levels of psoriasis model in Balb/c nude mice

Objective: To investigate the effect of Astragalus Injection on the Skin Lesion Degree and Caspase-14, SOCS1 and STAT3 Levels of Psoriasis Model in Balb/c Nude Mice. Methods:Sixty Balb/c nude mice were randomly divided into groups A, B and C with 20 mice in each group. Group A mice were used as blank control, group B mice as model group and group C mice as treatment group. The PASI score of psoriasis, skin thickness, inflammatory factors, serum levels of Caspase-14, SOCS1 and STAT3 in three groups of mice were analyzed after 2 weeks of treatment. Result: After treatment, the P ASI score of group B was significantly higher than that of group C, with statistical significance (P < 0.05);there was statistical significance in the measurements of lesion skin of three groups of mice after treatment (P <0.05). Compared with the blank control group, the thickness of lesion skin in group B and C was significantly higher, and the thickness of lesion skin in treatment group was significantly lower than that in control group (P < 0.05). Compared with the blank control group, the inflammatory factors IL-17, IL-22 and IL-23 in the B and C groups were significantly increased, and the inflammatory factors IL-17, IL-22 and IL-23 in the treatment group were significantly lower than those in the model group. The levels of serum C aspase-14, SOCS1 and STAT3 in three groups of mice were significantly different after treatment (P < 0.05). Compared with the blank control group, the levels of serum C aspase-14 and SOCS1 in B and C groups were significantly lower and the levels of STAT3 were significantly higher, and the levels of inflammatory factors aspase-14 and SO in treatment group were significantly higher than those in control group. The level of CS1 was significantly lower than that of model group, and the level of STAT3 was significantly higher than that of model group (P <0.05). Conclusion: Astragalus membranaceus injection can effectively improve the degree of psoriasis in Balb/c nude mice. Its possible mechanism is that it can decrease the expression of Caspase-14 and SOCS1, reduce the degree of keratosis in the lesion site of mice, improve the local surface hyperplasia, increase the level of STAT3 and enhance the level of local cell proliferation, which is of positive significance for the rehabilitation of psoriasis.

Knockdown of STAT3 by iRNA Inhibiting Migration and Invasion of Epithelial Ovarian Cancer Cells

Signal transducer and activator of transcription 3（STAT3） is a dual functional transcription factor with the functions of signal transduction and transcription regulation. It is reported that the expression of STAT3 in ovarian cancer is significantly higher and STAT3 can facilitate ovarian cancer growth and metastasis. To clarify the definite effect and molecular mechanism of STAT3 involved in ovarian cancer growth and metastasis, STAT3 expression was significantly downregulated by transfecting ovarian cancer model SK-OV-3 cells with the plasmid vector which express specific RNAi that targets human STAT3. The downregulated STAT3 not only decreased the invasion and migration but also inhibited the proliferation of SK-OV-3 cells. Western blot assay shows that the expression of vascular endothelial growth factor（VEGF） and that of Survivin were reduced in the cells with the plasma vector expressing specific RNAi that targets human STAT3. These results demonstrate that STAT3 involved in the invasion and migration of SK-OV-3 regulates the expression of VEGF and Survivin. In addition, VEGF and Survivin could play an important role in ovarian cancer growth and metastasis.

补阳还五汤通过调控PI3K/Akt、JAK2/STAT3信号促进BMSC趋化迁移对外伤性脊髓损伤大鼠神经元活性及认知功能的影响

目的研究补阳还五汤通过调控磷脂酰肌醇-3激酶/蛋白激酶B(PI3K/Akt)、内源性酪氨酸激酶(JAK)2/信号传导和转录启动因子(STAT)3信号促进骨髓间充质干细胞(BMSCs)趋化迁移对外伤性脊髓损伤大鼠的神经元活性及认知功能的影响。方法选取健康大鼠53只,随机分为健康组(健康大鼠常规饲养)、损伤组(建立脊髓损伤模型)、干预组(补阳还五汤治疗)、对照组(甲泼尼龙治疗),每组12只,剩余5只大鼠用于补阳还五汤含药血清制备。流式细胞术鉴定BMSCs细胞。Transwell小室法测大鼠BMSCs迁移。高架十字迷宫和Morris水迷宫实验检测大鼠认知功能。苏木素-伊红(HE)染色检测脊髓组织病理形态。TUNEL测脊髓组织神经细胞凋亡。免疫组化检测p-JAK2、p-STAT3。Western印迹测PI3K、p-PI3K、Akt、p-Akt。结果传代后的培养细胞呈旋窝状或放射状贴壁生长,细胞多呈星形、梭形或三角状,培养3代后,细胞贴壁加快、形态均一,呈旋窝状或单层放射状生长。培养细胞表面抗原CD29、CD90为阳性,CD31、CD45为阴性,提示其为BMSCs细胞。与健康组相比,损伤组总路程、进入开臂次数、穿越平台次数显著降低,不同时间的潜伏期显著升高(P<0.05)。与损伤组相比,干预组与对照组总路程、进入开臂次数、穿越平台次数显著升高,不同时间的潜伏期显著降低(P<0.05)。干预组与对照组各指标对比无统计学差异(P>0.05)。健康组脊髓组织结构完整。损伤组脊髓组织疏松水肿,有细胞空泡变性产生。相较于损伤组,干预组与对照组大鼠脊髓组织病理形态有所改善。与健康组相比,损伤组BMSCs、PI3K、Akt、p-PI3K、p-Akt显著降低,神经细胞凋亡率、p-JAK2、p-STAT3显著升高(P<0.05)。与损伤组相比,干预组BMSCs、PI3K、Akt、p-PI3K、p-Akt显著升高,神经细胞凋亡率、p-JAK2、p-STAT3显著降低(P<0.05)。干预组与对照组各指标水平无统计学差异(P>0.05)。结论补阳还五汤通过激活PI3K/Akt通路抑制JAK2/STAT3信号通路的激活,促进BMSCs的迁移,减轻神经细胞的凋亡,起到神经保护的作用,从而改善脊髓损伤大鼠的认知功能。

敲低NOD2通过调控(p)NF-κB/STAT3改善肝细胞癌细胞炎症反应

目的 探讨核苷酸结合寡聚化结构域蛋白2(NOD2)对肝细胞癌(HCC)细胞炎症反应的影响和机制。方法 培养HCC细胞。Western blot检测NOD2在HCC细胞和正常肝细胞中的表达水平。利用小干扰RNA(siRNA)建立NOD2的敲低RNA(siNOD2组)用来抑制NOD2的表达,阴性对照为siNC组,使用这二者处理HCC细胞。CCK-8法检测细胞增殖率,流式细胞术检测细胞凋亡率。Western blot检测NF-κB P65、STAT3、NOD2的表达,并检测磷酸化的(p-)NF-κB P65以及磷酸化的(p-)STAT3的表达水平。ELISA法测定培养HCC细胞培养物上清液中TNF-α、IL-12、IL-6、IFN-γ质量浓度的变化。在siNOD2处理HCC细胞的基础上,用NF-κB/STAT3的激活剂重组人Lipocalin-2(rhLipocalin-2)蛋白进行处理(siNOD2+rhLipocalin-2组),检测细胞增殖率、凋亡率和炎症因子水平。结果 与正常肝细胞相比,HCC细胞中NOD2的表达水平显著上调(P<0.05)。与siNC组相比,siNOD2组的NOD2、p-NF-κB P65及p-STAT3的表达水平均显著下调、细胞增殖率降低,细胞凋亡率增高,且细胞培养物上清液中TNF-α、IL-12、IL-6、IFN-γ水平均下调(均P<0.05)。而与siNOD2组相比,siNOD2+rhLipocalin-2组的p-NF-κB P65及p-STAT3的表达水平均显著上调,细胞增殖率增高,细胞凋亡率降低,且细胞培养物上清液中TNF-α、IL-12、IL-6、IFN-γ水平均上调(均P<0.05)。结论 敲低NOD2通过调控NF-κB/STAT3通路抑制HCC细胞的炎症反应,该结果为HCC治疗提供了一个新的潜在靶点。

FGF2对矿化诱导下STAT3介导的h DPSCs成牙分化作用研究

目的探讨成纤维细胞生长因子2(FGF2)对人牙髓干细胞(hDPSCs)成牙分化的影响及对信号转导子与激活子3(STAT3)通路的调节作用。方法hDPSCs随机分为对照组、FGF2组、Stattic组和FGF2+Stattic组,细胞进行成骨诱导。FGF2组细胞20 ng/mL FGF2处理,Stattic组细胞4μM Stattic预处理30 min,FGF2+Stattic组细胞4μM Stattic预处理30 min后,20 ng/mL FGF2处理。CCK-8检测细胞活力,qRT-PCR检测牙本质基质蛋白1(DMP-1)和牙本质涎磷蛋白(DSPP)mRNA相对表达量,检测碱性磷酸酶(ALP)活性,茜素红染色观察矿化情况,蛋白印迹法检测FGF2、p-STAT3、DMP-1和DSPP蛋白相对表达量。结果与对照组比较,FGF2组细胞增殖活性、ALP活性、DMP-1和DSPP mRNA表达量、FGF2、p-STAT3、DMP-1和DSPP蛋白表达量升高(P<0.05);与对照组比较,Stattic组细胞增殖活性和ALP活性减弱、DMP-1和DSPP mRNA表达量以及FGF2、p-STAT3、DMP-1和DSPP蛋白表达量降低(P<0.05);与FGF2组比较,FGF2+Stattic组细胞增殖活性和ALP活性减弱,DMP-1和DSPP mRNA和蛋白相对表达量、FGF2和p-STAT3蛋白相对表达量降低(P<0.05);与Stattic组比较,FGF2+Stattic组细胞增殖活性和ALP活性增强,DMP-1和DSPP mRNA和蛋白相对表达量、FGF2和p-STAT3蛋白相对表达量升高(P<0.05)。结论FGF2可诱导hDPSCs成牙本质分化,可能是通过调节STAT3信号通路发挥作用。