Mesalazine alleviated the symptoms of spontaneous colitis in interleukin-10 knockout mice by regulating the STAT3/NF-κB signaling pathway

BACKGROUND Excessive endoplasmic reticulum(ER)stress in intestinal epithelial cells can lead to damage to the intestinal mucosal barrier,activate the signal transducer and activator of transcription 3(STAT3)/nuclear factor kappa B(NF-κB)signaling pathway,and exacerbate the inflammatory response,thus participating in the pathogenesis of ulcerative colitis(UC).Mesalazine is a commonly used drug in the clinical treatment of UC.However,further studies are needed to determine whether mesalazine regulates the ER stress of intestinal epithelial cells,downregulates the STAT3/NF-κB pathway to play a role in the treatment of UC.AIM To study the therapeutic effects of mesalazine on spontaneous colitis in interleukin-10(IL-10)-/-mice.METHODS The 24-week-old IL-10-/-mice with spontaneous colitis were divided into the model group and the 5-amino salicylic acid group.Littermates of wild-type mice of the same age group served as the control.There were eight mice in each group,four males and four females.The severity of symptoms of spontaneous colitis in IL-10-/-mice was assessed using disease activity index scores.On day 15,the mice were sacrificed.The colon length was measured,and the histopathological changes and ultrastructure of colonic epithelial cells were detected.The protein expressions of STAT3,p-STAT3,NF-κB,IκB,p-IκB,and glucoseregulated protein 78 were identified using Western blotting.The STAT3 and NF-κB mRNA expressions were identified using real-time polymerase chain reaction.The glucose-regulated protein 78 and C/EBP homologous protein expressions in colon sections were detected using immunofluorescence.RESULTS Mesalazine reduced the symptoms of spontaneous colitis in IL-10 knockout mice and the histopathological damage of colonic tissues,and alleviated the ER stress in epithelial cells of colitis mice.Western blotting and quantitative real-time polymerase chain reaction results showed that the STAT3/NF-κB pathway in the colon tissue of model mice was activated,suggesting that this pathway was involved in the pathogenesis of UC and might become a potential therapeutic target.Mesalazine could down-regulate the protein expressions of p-STAT3,NF-κB and p-IκB,and down-regulate the mRNA expression of STAT3 and NF-κB.CONCLUSION Mesalazine may play a protective role in UC by reducing ER stress by regulating the STAT3/NF-κB signaling pathway.

特发性间质性肺炎病人血清STAT3和FOXM1水平与病情程度及预后的关系研究

目的探讨特发性间质性肺炎(IIP)病人血清信号转导及转录激活因子3(STAT3)、叉头框转录因子M1(FOXM1)表达水平与其病情程度和预后的关系。方法选取2017年1月至2022年10月四川省建筑医院收治的298例IIP病人为IIP组,选取同时期该院收治的298例普通型间质性肺炎(UIP)病人为UIP组,另选取同时期在该院体检的280例健康人员为对照组。采用酶联免疫吸附测定(ELISA)检测血清STAT3与FOXM1表达水平,并采用全肺纤维化高分辨率CT(HRCT)评分评价IIP病情严重程度。分析三组受试者血清STAT3与FOXM1表达水平差异。对IIP组病人随访3个月,根据其生存情况分为预后良好组与预后不良组,分析预后不良与预后良好组IIP病人血清STAT3与FOXM1表达水平差异,并采用Spearman法分析IIP病人血清STAT3与FOXM1表达水平与全肺纤维化HRCT评分之间的关系。采用logistic回归分析IIP病人预后不良的影响因素,绘制受试者操作特征曲线(ROC曲线)评估血清STAT3与FOXM1表达水平对IIP病人预后不良的预测效能。结果IIP组、UIP组、对照组血清STAT3[(1.50±0.39)ng/L、(1.32±0.31)ng/L、(1.01±0.27)ng/L]、FOXM1[(34.56±5.64)ng/L、(22.69±4.11)ng/L、(15.51±3.94)ng/L]水平均依次降低(P<0.05)。IIP病人随访3个月共有41例病人死亡,预后不良发生率为13.76%(41/298);预后不良组IIP病人血清STAT3、FOXM1水平均高于预后良好组(P<0.05)。Spearman相关性分析表明,IIP病人血清STAT3、FOXM1水平均与全肺纤维化HRCT评分正相关(rs=0.65,rs=0.57;P<0.001)。另预后不良组年龄、Ⅱ型肺泡细胞表面抗原(KL-6)、白细胞计数、红细胞沉降率(ESR)、C反应蛋白(CRP)水平、全肺纤维化HRCT评分均高于预后良好组(P<0.05);第一秒用力呼气量(FEV1)、最大自主通气量(MVV)均低于预后良好组(P<0.05)。多因素logistic回归分析显示,年龄、KL-6、白细胞、ESR、CRP、STAT3、FOXM1水平、全肺纤维化HRCT评分均是导致IIP病人预后不良的危险因素(P<0.05),FEV1、MVV为保护因素(P<0.05)。ROC分析结果表明,血清STAT3、FOXM1水平联合预测IIP病人预后的灵敏度高于STAT3、FOXM1单独预测的灵敏度(χ^(2)=8.10、6.12,P=0.002、0.008);联合预测的曲线下面积(AUC)高于STAT3、FOXM1单独预测的AUC(Z=3.15、2.54,P=0.002、0.011)。结论IIP病人血清STAT3、FOXM1表达水平均上升,二者与IIP病情程度正相关,均对IIP病人预后具有良好的预测价值,且联合预测效能更高。

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.

Apigenin ameliorates imiquimod-induced psoriasis in C57BL/6J mice by inactivating STAT3 and NF-κB

Psoriasis is a chronic autoimmune disease featured by patches on the skin.It is caused by malfunction of immune cells and keratinocytes with inflammation as one of its key features.Apigenin(API)is a natural flavonoid with anti-inflammatory and immunoregulatory properties.Therefore,we speculated that API can ameliorate psoriasis,and determined its effect on the development of psoriasis by using imiquimod(IMQ)-induced psoriasis mouse model.Our results showed that API attenuated IMQ-induced phenotypic changes,such as erythema,scaling and epidermal thickening,and improved splenic hyperplasia.Abnormal differentiation of immune cells was restored in API-treated mice.Mechanistically,we revealed that API is a key regulator of signal transducer activator of transcription 3(STAT3).API regulated immune responses by reducing interleukin-23(IL-23)/STAT3/IL-17A axis.Moreover,it suppressed IMQ-caused cell hyperproliferation by inactivating STAT3 through regulation of extracellular signal-regulated kinase 1/2 and nuclear factor-κB(NF-κB)pathway.Furthermore,API reduced expression of inflammatory cytokines through inactivation of NF-κB.Taken together,our study demonstrates that API can ameliorate psoriasis and may be considered as a strategy for psoriasis treatment.

山柰酚调节IL-6/STAT3信号通路对妊娠糖尿病大鼠炎症反应的影响

目的探讨山柰酚(kaempferol,Kpf)调节白细胞介素-6(interleukin-6,IL-6)/信号转导与转录激活子3(signal transducer and activator of transcription 3,STAT3)信号通路对妊娠糖尿病(gestational dia-betes mellitus,GDM)大鼠炎症反应的影响。方法雌雄同笼与高脂高糖喂养联合腹腔注射链脲佐菌素(streptozotocin,STZ)构建GDM大鼠模型,将造模成功大鼠按照随机数字表达分为Model组、Kpf低、中、高剂量组、激活剂组(IL-6/STAT3通路激活剂rIL-6),每组12只,同批12只妊娠大鼠作为control组,各组腹腔注射相应药物,每天1次,直到妊娠19 d。罗氏血糖仪测量大鼠空腹血糖(fasting blood glucose,FBG);分析各组大鼠妊娠结局;ELISA法测定大鼠血清空腹胰岛素(fasting insulin,FINS)及胎盘中白细胞介素(interleukin,IL)-6、IL-1β、肿瘤坏死因子(tumor necrosis factor-α,TNF-α)水平;苏木精伊红(hematoxylin-eosin,HE)染色观察大鼠胎盘组织中病理损伤;TUNEL染色检测胎盘组织细胞凋亡;蛋白质印迹测定胎盘中IL-6/STAT3通路蛋白表达。结果与control组比较,Model组大鼠FBG、FINS、HOMA-IR、胎鼠质量、IL-6、IL-1β、TNF-α水平、胎盘组织IL-6、p-STAT3/STAT3水平及细胞凋亡率增加,活胎率、窝产子数下降(P<0.05);相较于Model组,Kpf低、中、高剂量组大鼠FBG、FINS、HOMA-IR、胎鼠质量、IL-6、IL-1β、TNF-α水平、胎盘组织IL-6、p-STAT3/STAT3水平及细胞凋亡率下降,活胎率、窝产子数增加(P<0.05);相较于Kpf高剂量组,激活剂组上述指标变化均显著逆转(P<0.05)。结论Kpf可能通过抑制IL-6/STAT3信号通路降低GDM大鼠胎盘炎症,减轻胰岛素抵抗。

带状疱疹患者血清STAT3,LncRNA XIST水平表达与疱疹后遗神经痛发生的相关性研究

目的 探讨信号转导和转录激活因子3(signal transduction and transcription activator 3,STAT3)、长链非编码RNA X染色体失活特异转录物(long non-coding RNA X chromosome inactivation specific transcript,LncRNA XIST)在带状疱疹后遗神经痛(postherpetic neuralgia,PHN)患者中的表达水平及其与PHN病情的相关性。方法 回顾性选取绵阳市中心医院2021年4月~2023年3月收治的179例带状疱疹患者为研究对象,根据PHN的发生与否分为PHN组(n=43)和非PHN组(n=136),并根据疼痛程度将43例PHN患者分为轻度组(n=20)、中度组(n=16)和重度组(n=7);另选取绵阳市中心医院同期健康体检者120例作为对照组。血清STAT3水平采用ELISA进行检测;LncRNA XIST水平采用RT-qPCR进行检测;采用多因素Logistic回归分析法分析影响PHN发生的因素;采用受试者工作特征(ROC)曲线分析血清STAT3和LncRNA XIST水平对PHN发生的预测价值。结果 与对照组比较,非PHN组、PHN组血清STAT3(1.44±0.38 ng/L,2.13±0.51 ng/L vs 0.86±0.20 ng/L),LncRNA XIST(1.79±0.47,2.82±0.66 vs 1.05±0.24)水平均明显升高(t=14.986,15.549;22.894,25.197),且PHN组高于非PHN组(t=9.514,11.292),差异具有统计学意义(均P<0.05)。与轻度组比较,中度组和重度组血清STAT3(2.21±0.53ng/L,3.04±0.59 ng/L vs 1.72±0.42 ng/L)和LncRNA XIST(2.96±0.68,4.15±0.76 vs 2.24±0.51)水平显著升高(t=3.097,3.632;6.443,7.500),且重度组显著高于中度组(t=3.343,3.731),差异具有统计学意义(均P<0.05)。与非PHN组比较,PHN组辅助性T细胞(T helper cell 17,Th17)/调节性T细胞(regulatory T cells,Treg)(0.60±0.12 vs 0.54±0.08)和白细胞介素-10(interleukin-10,IL-10)水平(29.69±4.58 pg/ml vs 23.57±3.62 pg/ml)明显升高,差异具有统计学意义(t/χ^(2)=3.765,9.040,均P<0.05)。多因素Logistic回归分析显示,STAT3,LncRNA XIST,Th17/Treg,IL-10均为影响PHN发生的独立危险因素(OR=1.157,1.149,1.153,1.125,均P<0.05);ROC曲线结果显示,血清STAT3,LncRNA XIST水平单独预测PHN发生的AUC(95%CI)分别为0.807(0.741~0.862),0.787(0.720~0.845),血清STAT3和LncRNA XIST联合预测PHN发生的AUC(95%CI)为0.881(0.824~0.924),高于二者单独预测的AUC,差异具有统计学意义(Z=1.994,2.175,P=0.046,0.030),敏感度和特异度分别为81.82%,84.56%。结论 STAT3和LncRNA XIST在PHN患者血清中升高,PHN患者疼痛程度越高STAT3和LncRNA XIST水平越高,二者联合检测对PHN的预测价值较高,有一定临床参考价值。

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.

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 tumorbearing nude mice in vivo.METHODS： To construct the recombinant plasmid of pSilencer 3.0-H1-STAT3-siRNA-GFP （pSHI-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 pSHI-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 （RT- PCR）. 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 ceils 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.

STAT3 activation of tumor-associated macrophages is associated with cytokines of tumor microenvironment and prognostic factors in breast cancer

Objective:The aim of this study was to investigate the correlation of STAT3 activation of tumor-associated macrophages(TAMs) and local cytokines(IL-1β,TNF-α,TGF-β and IL-12) and prognostic factors in breast cancer.Methods:TAMs in 50 primary breast cancers and macrophages in 15 normal breasts were examined by immunohistochemistry.And STAT3 DNA-binding activity of TAMs in 33/50 primary breast cancers was measured by transcription factor DNA-binding ELISA.In addition,the concentrations of IL-1β,TNF-α,TGF-β and IL-12 were measured in the 33 primary breast cancers extracts by ELISA.The correlation between STAT3 activity of TAMs and concentrations of IL-1β,TNF-α,TGF-β and IL-12 were analyzed.The correlation between STAT3 activity of TAMs and conventional clinicopathologic parameters were also evaluated.Results:The macrophages density showed a significant increase in primary breast cancers compared to normal breasts.STAT3 DNA-binding activity of TAMs in breast cancer was significantly higher than that of monocytes/macrophages from peripheral blood of the patients.Furthermore,STAT3 activity of TAMs was correlated significantly with the levels of IL-1β,TNF-α and TGF-β in breast cancer tissues.But an inverse association was observed between STAT3 activity of TAMs and IL-12.In addition,STAT3 activity of TAMs was higher in high histological type than in low histological type,and STAT3 activity of TAMs was higher in CerBb-2 positive than CerBb-2 negative.Conclusion:STAT3 activation of TAMs may be associate with increasing of IL-1β,TNF-α and TGF-β and decreasing of IL-12 in breast cancer.STAT3 activation of TAMs may also be correlated with histological grade and CerBb-2 status of breast cancer.

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

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

青藤碱调节CXCR4-STAT3轴对卵巢癌A2780细胞增殖、迁移和血管生成拟态的影响

目的:探讨青藤碱(Sinomenine)对卵巢癌细胞增殖、迁移和血管生成拟态的影响及作用机制。方法:使用不同浓度(0、0.25、0.50、1.0、2.0、4.0、8.0 mmol/L)的青藤碱分别处理A2780细胞24、48 h, CCK-8法检测细胞存活率。将A2780细胞随机分为对照(Control)组、CXCR4激活剂基质细胞衍生因子-1(SDF-1)(SDF-1,100 ng/mL)组、CXCR4抑制剂普乐沙福(Plerixafor, 500 ng/mL)组、青藤碱(Sinomenine, 1.0 mmol/L)组、Sinomenine+SDF-1(1.0 mmol/L Sinomenine+100 ng/mL SDF-1)组,分别检测A2780细胞增殖、迁移与侵袭,体外血管生成拟态形成实验检测青藤碱对血管生成拟态的影响,Western blot法检测血管内皮生长因子(VEGF)、上皮细胞激酶(EphA2)、基质金属蛋白酶9(MMP-9)、MMP-2、CXC趋化因子受体4(CXCR4)、信号转导与转录激活因子3(STAT3)、磷酸化STAT3(p-STAT3)蛋白表达。结果:青藤碱可有效抑制A2780细胞增殖,且呈浓度依赖性;青藤碱可显著抑制A2780细胞迁移、侵袭及血管生成拟态形成,并下调血管生成拟态标志蛋白VEGF、EphA2、MMP-9、MMP-2表达,抑制CXCR4、p-STAT3表达(P<0.05),青藤碱的这一抑制作用可被CXCR4激活剂减弱。结论:青藤碱可能通过抑制CXCR4-STAT3轴,抑制卵巢癌细胞增殖、迁移、侵袭及血管生成拟态形成。

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。

重症肺炎患者血清JAK2/STAT3信号通路、TLR表达及感染病原菌与耐药性

目的分析重症肺炎患者血清Janus激酶2/信号转导与转录激活子3(JAK2/STAT3)信号通路、Toll样受体(TLR)表达及感染病原菌特点与耐药性。方法选取2020年11月—2023年11月长沙市第四医院收治的重症肺炎患者97例为研究组,同期普通肺炎患者98例为对照组,分析重症肺炎患者感染病原菌分布及其耐药性,比较两组血清JAK2、STAT3、TLR2及TLR4表达情况;通过受试者工作特征(ROC)曲线分析四者单独及联合检测对重症肺炎的预测价值。结果97例重症肺炎患者共分离出病原菌108株,其中革兰阴性菌74株占68.52%;主要革兰阴性菌鲍氏不动杆菌对哌拉西林和头孢他啶耐药性较强,耐药率均>70%,对米诺环素、黏菌素、替加环素较敏感,耐药率<10%;铜绿假单胞菌对哌拉西林耐药性较强,耐药率为34.62%,对阿米卡星、米诺环素和妥布霉素的耐药率<10%,对黏菌素敏感;主要革兰阳性菌金黄色葡萄球菌对青霉素耐药性较强,耐药率为90.48%,对利奈唑胺、万古霉素、替考拉宁均敏感;研究组JAK2、STAT3、TLR2及TLR4水平均高于对照组(P<0.05),绘制ROC曲线获得JAK2、STAT3、TLR2及TLR4联合诊断重症肺炎的曲线下面积(AUC)为0.930,敏感度和特异度分别为78.40%,92.90%。结论重症肺炎感染检出革兰阴性菌占比较高,鲍氏不动杆菌、金黄色葡萄球菌对常用抗菌药物耐药率较高,重症肺炎发生后JAK2/STAT3信号通路激活,且TLR2、TLR4表达升高,联合检测有助于诊断重症肺炎。

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。

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。

川崎病合并支原体感染病儿瓣膜炎性损伤与IL-6/STAT3信号通路表达变化的关系分析

目的探讨川崎病(KD)合并支原体感染病儿瓣膜炎性损伤与白细胞介素-6(IL-6)/信号转导及转录激活蛋白3(STAT3)信号通路表达变化的关系。方法前瞻性选取2018年1月至2022年12月安阳市妇幼保健院收治的50例KD合并肺炎支原体(MP)(KD-MP)感染病儿为感染组,另选取同期收治的106例单纯KD病儿为未感染组,比较感染组与未感染组病儿瓣膜炎性损伤情况。再根据KD病儿是否伴有冠状动脉损伤(CAL)损伤分为CAL组(n=84)、无CAL组(n=72),比较两组病儿IL-6和STAT3表达水平,通过多因素logistic回归分析KD病儿发生CAL的危险因素。比较KD并CAL不同预后病儿IL-6和STAT3表达变化,使用受试者操作特征曲线(ROC曲线)分析IL-6、STAT3水平对KD并CAL病儿预后的预测价值。结果感染组KD病儿发热时间、住院时间、初诊CAL发生率均高于未感染组(P<0.05);感染组KD病儿C反应蛋白(CRP)、降钙素原(PCT)、红细胞沉降率(ESR)、IL-6 mRNA表达水平和STAT3 mRNA表达水平均高于未感染组[(51.32±12.62)mg/L比(36.58±10.98)mg/L,(1.33±0.35)μg/L比(1.21±0.31)μg/L,(84.69±15.66)mm/h比(49.62±16.43)mm/h,4.14±1.09比3.06±1.15,2.81±0.89比1.68±0.54,均P<0.05]。CAL组病儿IL-6、STAT3 mRNA表达水平高于无CAL组(4.13±1.22比2.56±0.87,2.46±0.71比1.55±0.44,均P<0.05);多因素回归分析结果显示,MP感染、IL-6 mRNA、STAT3 mRNA表达是KD病儿发生CAL的独立危险因素(均P<0.05)。KD并CAL预后良好组病儿IL-6、STAT3 mRNA表达水平低于预后不良组(3.57±1.06比4.57±1.13,1.94±0.58比2.87±0.87,均P<0.05),ROC曲线结果显示IL-6、STAT3及其联合预测KD并CAL预后的曲线下面积(AUC)分别为0.75、0.81、0.91,且联合检测的AUC高于IL-6、STAT3单独检测(均P<0.05)。结论KD合并MP病儿瓣膜炎性损伤更为严重,IL-6/STAT3信号通路活化是其危险因素,且该信号通路对KD并CAL病儿预后有较高的预测价值。

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。

鸢尾素调节JAK2/STAT3信号通路对牙周炎大鼠牙周组织损伤的影响

目的:探讨鸢尾素调节Janus蛋白酪氨酸激酶2(Janus protein tyrosine kinase 2,JAK2)/信号转导和转录激活子3(Signal transduction and activator of transcription 3,STAT3)信号通路对牙周炎大鼠牙周组织损伤的影响。方法:通过结扎和接种牙龈卟啉单胞菌液建立牙周炎大鼠模型,将大鼠随机分为模型组、鸢尾素低(鸢尾素-L,50 mg/kg)、中(鸢尾素-M,100 mg/kg)、高剂量(鸢尾素-H,200 mg/kg)组、鸢尾素-H+激活剂(200 mg/kg鸢尾素+2 mg/kg香豆霉素)组,每组10只,并以注射等体积生理盐水的正常大鼠对照组。干预结束后,对大鼠牙龈出血指数、牙齿松动度评分;牙槽骨吸收、肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)、白细胞介素(Interleukin,IL)-6、IL-1β以及丙二醛(Malondialdehyde,MDA)、超氧化物歧化酶(Superoxide dismutase,SOD)水平分别以Micro-CT试剂盒检测;HE检测牙周组织病理学变化;Western blot检测JAK2、STAT3、p-JAK2、p-STAT3蛋白表达。结果:与对照组相比,模型组大鼠牙周组织被破坏,炎性浸润严重,牙龈出血指数、牙齿松动度评分、牙槽骨吸收、TNF-α、IL-6、IL-1β、MDA水平、p-JAK2/JAK2、p-STAT3/STAT3表达显著增加,SOD水平显著降低(P<0.05);与模型组相比,不同剂量的鸢尾素组大鼠病理损伤得到改善,牙龈出血指数、牙齿松动度评分、牙槽骨吸收、TNF-α、IL-6、IL-1β、MDA水平、p-JAK2/JAK2、p-STAT3/STAT3表达显著降低,SOD水平显著增加,具有剂量依赖性(P<0.05);与鸢尾素-H组相比,鸢尾素-H组+激活剂组大鼠病理损伤加重,大鼠牙龈出血指数、牙齿松动度评分、牙槽骨吸收、TNF-α、IL-6、IL-1β、MDA水平、p-JAK2/JAK2、p-STAT3/STAT3表达显著增加,SOD水平显著降低(P<0.05)。结论:鸢尾素抑制牙周炎大鼠氧化应激、炎性反应,减轻大鼠牙周组织损伤,减少牙槽骨吸收,可能与抑制JAK2/STAT3信号通路有关。

大柴胡汤含药血清通过JAK2/STAT3信号通路抑制AR42J细胞炎症反应

目的探讨大柴胡汤含药血清对胰腺腺泡细胞炎症反应的防治作用及可能机制。方法HPLC法检测大柴胡汤中指标成分含量;采用雨蛙肽处理AR42J细胞复制急性腺泡细胞炎症模型,采用CCK-8方法检测大柴胡汤含药血清对细胞活力的影响;Western blot方法检测细胞内JAK2、STAT3及NFκB磷酸化程度;ELISA法检测IL-6和TNF-α含量。结果大柴胡汤中黄芩苷和芍药苷含量分别为13.02 mg/mL、7.19 mg/mL,大柴胡汤含药血清用量不超过20%;大柴胡汤含药血清降低炎性细胞上清液中TNF-α和IL-6含量,降低细胞内JAK2、STAT3及NFκB磷酸化作用,减轻细胞炎症反应;STAT3抑制剂cucurbitacin可减弱大柴胡汤的炎症抑制作用。结论大柴胡汤含药血清可减轻腺泡细胞急性炎症作用,其机制可能为抑制细胞内JAK2/STAT3信号通路进而降低NFκB活性,减少促炎性细胞因子合成。