黄芩苷对干酵母致热大鼠的解热作用及血清TNF-α、IL-1β、IL-6、PGE_(2)、cAMP和脑组织NF-κB表达的影响

目的:观察黄芩苷对干酵母致热大鼠的解热作用并探讨其作用机制。方法:采用背部皮下注射干酵母构建大鼠发热模型,SD雄性大鼠随机分为正常对照,模型组,阳性组(阿司匹林,0.1 g/kg),黄芩苷高、中、低剂量组(160、80、40 mg/kg),连续给药3 d,测定各组大鼠肛温的变化;酶联免疫法(ELISA)检测血清肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)、白细胞介素-6(IL-6)、前列腺素E_(2)(PGE_(2))与环磷酸腺苷(cAMP)水平;Western Blot检测各组大鼠脑组织NF-κB p65(核转录因子-κB p65)蛋白表达。结果:黄芩苷高剂量组有显著解热效果(P<0.01),黄芩苷各剂量组均可不同程度降低大鼠血清TNF-α、IL-1β、IL-6、PGE 2和cAMP含量;与正常组比较,模型组脑组织NF-κB p65蛋白表达增多,黄芩苷高剂量组可明显降低大鼠脑组织NF-κB p65表达(P<0.05)。结论:黄芩苷可显著性降低干酵母引起的体温升高,解热机制可能与抑制TNF-α、IL-1β、IL-6、PGE_(2)与cAMP的分泌和减少脑组织NF-κB p65蛋白表达有关。

支气管肺泡灌洗液及血清NF-κB、TLR-2、TLR-4、TNF-α在肺炎诊断中的应用价值

目的探讨支气管肺泡灌洗液(BALF)及血清核转录因子-κB(NF-κB)、Toll样受体2(TLR-2)、Toll样受体4(TLR-4)、肿瘤坏死因子α(TNF-α)在肺炎诊断中的应用价值。方法选取2023年1月—2024年3月于吉林大学第二医院就诊的57例肺炎患者纳入研究组,另取同期体检的健康者57例纳入对照组,均采集外周静脉血与肺泡灌洗液,检测NF-κB、TLR-2、TLR-4、TNF-α水平,采用Pearson相关性分析患者血清及BALF中NF-κB、TLR-2、TLR-4、TNF-α的相关性。结果研究组患者血清NF-κB(41.74±4.75)μg/L、TLR-2(5.32±1.12)ng/L、TLR-4(6.44±1.61)ng/L、TNF-α(4.36±1.03)ng/L水平高于对照组,差异有统计学意义(P<0.05)。研究组患者BALF中NF-κB(8.19±2.62)μg/L、TLR-2(2.97±0.79)ng/L、TLR-4(4.24±1.18)ng/L、TNF-α(2.04±0.59)ng/L水平高于对照组,差异有统计学意义(P<0.05)。Pearson相关性分析显示,患者血清及BALF中NF-κB、TLR-2、TLR-4、TNF-α水平呈正相关性(P<0.05)。结论肺炎患者血清及BALF中NF-κB、TLR-2、TLR-4、TNF-α均呈高表达,联合检测有助于早期诊断及评估病情。

TNF-α/NF-κB信号通路在慢性阻塞性肺疾病中的作用及中医药调控研究进展

慢性阻塞性肺疾病(COPD)是一种常见的呼吸系统疾病,发病率和死亡率很高,显著影响着患者的生存质量。临床现有的药物治疗方案因其副作用多、疗效欠佳等原因给患者带来了沉重的身心和经济负担。同时,COPD的具体机制仍不明确,这也给治疗带来了一定的难度。因此,阐明其潜在机制,提出安全、有效的治疗方案,是目前临床治疗COPD亟需解决的问题。大量研究证明,肿瘤坏死因子-α(TNF-α)/核转录因子-κB(NF-κB)信号通路在COPD的发生发展中发挥着至关重要的作用;中医药在防治COPD具有多靶点、多成分、副作用低等特点,其中有不少靶向调控TNF-α/NF-κB信号通路的中药复方和中药有效成分。本文综述近几年TNF-α/NF-κB信号通路对COPD的调控作用以及中医药靶向调控TNF-α/NF-κB信号通路防治COPD的研究进展,以期为临床治疗COPD提供一定线索。

Downregulation of MUC1 Inhibits Proliferation and Promotes Apoptosis by Inactivating NF-κB Signaling Pathway in Human Nasopharyngeal Carcinoma

Objective To investigate the effect of mucin 1(MUC1)on the proliferation and apoptosis of nasopharyngeal carcinoma(NPC)and its regulatory mechanism.Methods The 60 NPC and paired para-cancer normal tissues were collected from October 2020 to July 2021 in Quanzhou First Hospital.The expression of MUC1 was measured by real-time quantitative PCR(qPCR)in the patients with PNC.The 5-8F and HNE1 cells were transfected with siRNA control(si-control)or siRNA targeting MUC1(si-MUC1).Cell proliferation was analyzed by cell counting kit-8 and colony formation assay,and apoptosis was analyzed by flow cytometry analysis in the 5-8F and HNE1 cells.The qPCR and ELISA were executed to analyze the levels of TNF-αand IL-6.Western blot was performed to measure the expression of MUC1,NFкB and apoptosis-related proteins(Bax and Bcl-2).Results The expression of MUC1 was up-regulated in the NPC tissues,and NPC patients with the high MUC1 expression were inclined to EBV infection,growth and metastasis of NPC.Loss of MUC1 restrained malignant features,including the proliferation and apoptosis,downregulated the expression of p-IкB、p-P65 and Bcl-2 and upregulated the expression of Bax in the NPC cells.Conclusion Downregulation of MUC1 restrained biological characteristics of malignancy,including cell proliferation and apoptosis,by inactivating NF-κB signaling pathway in NPC.

Apatinib reduces liver cancer cell multidrug resistance by modulating NF-κB signaling pathway

Objectives:This investigation aimed to elucidate the inhibitory impact of apatinib on the multidrug resistance of liver cancer both in vivo and in vitro.Methods:To establish a Hep3B/5-Fu resistant cell line,5-Fu concentrations were gradually increased in the culture media.Hep3B/5-Fu cells drug resistance and its alleviation by apatinib were confirmed via flow cytometry and Cell Counting Kit 8(CCK8)test.Further,Nuclear factor kappa B(NF-κB)siRNA was transfected into Hep3B/5-Fu cells to assess alterations in the expression of multidrug resistance(MDR)-related genes and proteins.Nude mice were injected with Hep3B/5-Fu cells to establish subcutaneous xenograft tumors and then categorized into 8 treatment groups.The treatments included oxaliplatin,5-Fu,and apatinib.In the tumor tissues,the expression of MDRrelated genes was elucidated via qRT-PCR,immunohistochemistry,and Western blot analyses.Results:The apatinibtreated mice indicated slower tumor growth with smaller size compared to the control group.Both the in vivo and in vitro investigations revealed that the apatinib-treated groups had reduced expression of MDR genes GST-pi,LRP,MDR1,and p-p65.Conclusions:Apatinib effectively suppresses MDR in human hepatic cancer cells by modulating the expression of genes related to MDR,potentially by suppressing the NF-κB signaling pathway.

Kuicolong-yu enema decoction retains traditional Chinese medicine enema attenuates inflammatory response ulcerative colitis through TLR4/NF-κB signaling pathway

BACKGROUND Ulcer colitis(UC)is a chronic,nonspecific,and noninfectious inflammatory bowel disease.Recently,Toll-like receptors(TLRs)have been found to be closely associated with clinical inflammatory diseases.Achieving complete remission in patients with intermittent periods of activity followed by dormancy is challenging.Moreover,no study has explored the mechanism by which Kuicolong-yu enema decoction retains traditional Chinese medicine enemas to attenuate the inflammatory response in UC.AIM To explore the mechanism by which Kuicolong-yu enema decoction retains traditional Chinese medicine enemas to attenuate the inflammatory response in UC.METHODS This prospective clinical study included patients who met the exclusion criteria in 2020 and 2021.The patients with UC were divided into two groups(control and experimental).The peripheral blood of the experimental and control groups were collected under aseptic conditions.The expression of TLR4 protein,NF-κB,IL-6,and IL-17 was detected in the peripheral blood of patients in the experimental group and control group before and 1 month after taking the drug.Linear co rrelation analysis was used to analyze the relationship between the expression level of TLR4 protein and the expression levels of downstream signal NF-κB and inflammatory factors IL-6 and IL-17,and P<0.05 was considered statistically significant.RESULTS There were no significant differences in the patient characteristics between the control and experimental groups.The results showed that the expression levels of TLR4 and NF-κB in the experimental group were significantly lower than those in the control group(P<0.05).The levels of IL-6 and IL-17 in the experimental group were significantly lower than those in the control group(P<0.05).The TLR4 protein expression in the experimental group was positively correlated with the expression level of downstream signal NF-κB and was positively correlated with the levels of downstream inflammatory cytokines IL-6 and IL-17(r=0.823,P<0.05).CONCLUSION Kuicolong-yu enema decoction retains traditional Chinese medicine enema attenuates the inflammatory response of UC through the TLR4/NF-κB signaling pathway.

Parthenolide enhances the metronomic chemotherapy effect of cyclophosphamide in lung cancer by inhibiting the NF-kB signaling pathway

BACKGROUND Parthenolide(PTL),a sesquiterpene lactone derived from the medicinal herb Chrysanthemum parthenium,exhibits various biological effects by targeting NF-kB,STAT3,and other pathways.It has emerged as a promising adjunct therapy for multiple malignancies.AIM To evaluate the in vitro and in vivo effect of PTL on cyclophosphamide(CTX)metronomic chemotherapy.METHODS The cytotoxicity of PTL and CTX on Lewis lung cancer cells(LLC cells)was assessed by measuring cell activity and apoptosis.The anti-tumor efficiency was evaluated using a tumor xenograft mice model,and the survival of mice and tumor volume were monitored.Additionally,the collected tumor tissues were analyzed for tumor microenvironment indicators and inflammatory factors.RESULTS In vitro,PTL demonstrated a synergistic effect with CTX in inhibiting the growth of LLC cells and promoting apoptosis.In vivo,metronomic chemotherapy com-bined with PTL and CTX improved the survival rate of tumor-bearing mice and reduced tumor growth rate.Furthermore,metronomic chemotherapy combined with PTL and CTX reduced NF-κB activation and improved the tumor immune microenvironment by decreasing tumor angiogenesis,reducing Transforming growth factorβ,andα-SMA positive cells.CONCLUSION PTL is an efficient compound that enhances the metronomic chemotherapy effects of CTX both in vitro and in vivo,suggesting its potential as a supplementary therapeutic strategy in metronomic chemotherapy to improve the chemotherapy effects.

Aszonapyrone A Isolated from Neosartorya spinosa IFM 47025 Inhibits the NF-κB Signaling Pathway Activated by Expression of the Ependymoma-Causing Fusion Protein ZFTA-RELA

Ependymoma is a rare and chemotherapy-resistant brain tumor, which has resulted in a delay in the development of drugs to treat it. A subclass of supratentorial ependymomas (ST-EPN), designated ST-EPN-zinc finger-translocation-associated (ZFTA, ST-EPN-ZFTA), exhibits the expression of a fusion protein comprising ZFTA and v-rel reticuloendotheliosis viral oncogene homolog A (RELA), an effector transcription factor of the nuclear factor-kappa B (NF-κB) pathway (ZFTA-RELA). The expression of ZFTA-RELA results in the hyperactivation of the oncogenic NF-κB signaling pathway, which ultimately leads to the development of ST-EPN-ZFTA. To identify inhibitors of the NF-κB signaling pathway activated by the expression of ZFTA-RELA, we used a doxycycline-inducible ZFTA-RELA-expressing NF-κB reporter cell line and found that extracts of the fungus Neosartorya spinosa IFM 47025 exhibited NF-κB inhibitory activity. We identified eight compounds [aszonapyrone A (2), sartorypyrone A (3), epiheveadride (4), acetylaszonalenin (5), (R)-benzodiazepinedione (6), aszonalenin (7), sartorypyrone E (8) and (Z, Z)-N,N’-(1,2-bis[(4-methoxyphenyl)methylene]-1,2-ethanediyl)bis-formamide (9)] from N. spinosa IFM 47025 culture extract using a variety of chromatographic techniques. The structures of these compounds were identified through the analysis of various instrumental data (1D, 2D-NMR, MS, and optical rotation). The NF-κB responsive reporter assay indicated that compounds 2, 3, 5, 7, and 9 exhibited inhibitory activity. We further evaluated the inhibitory activity of these compounds against the expression of endogenous NF-κB responsive genes (CCND1, L1CAM, ICAM1, and TNF) and found that compound 2 showed significant inhibitory activity. Further studies are required to elucidate the mechanism of action of compound 2, which may serve as a lead compound for the development of a novel therapy for ST-EPN-ZFTA.

TNF-α调节NF-κB/PXR炎症途径对肺癌细胞生物行为的影响及作用机制

目的分析TNF-α调节NF-κB/PXR炎症途径对肺癌细胞生物行为的影响及作用机制。方法将A549细胞分为对照组和干预组,对照组正常培养不做任何干预处理,干预组使用人TNF-α以10 ng/ml浓度共培养12 h;检测细胞增殖、细胞周期、细胞凋亡、细胞侵袭活性;并采用PCR及WB法检测细胞中NF-κB、PXR蛋白和mRNA表达水平。结果在TNF-α干预24 h、48 h和72 h时,干预组细胞检测的吸光度值明显高于对照组,且差异存在统计学意义(P<0.05);采用TNF-α干预24 h后干预组A549细胞的凋亡率明显低于对照组,且差异存在统计学意义(P<0.05);干预24 h后采用TNF-α处理的干预组细胞的S期比例明显高于对照组,而G_(2)+M期和G_(0)+G_(1)期细胞比例明显低于对照组,且差异存在统计学意义(P<0.05);干预24 h后采用TNF-α处理的干预组细胞侵袭数明显高于对照组,且差异存在统计学意义(P<0.05);干预24 h后采用TNF-α处理的干预组NF-κB、PXR蛋白表达水平均明显高于对照组,且差异存在统计学意义(P<0.05);干预24 h后采用TNF-α处理的干预组细胞NF-κB及PXR mRNA表达均明显高于对照组,且差异存在统计学意义(P<0.05)。结论TNF-α可通过调控NF-κB/PXR炎症途径实现对肺癌细胞增殖、凋亡和侵袭的调控。

基于TLR-4/MyD88/NF-κB对健脾化滞丸治疗溃疡性结肠炎的机制进行拆方研究

目的:通过建立脾虚湿蕴型溃疡性结肠炎(UC)大鼠模型,探讨健脾化滞丸及其不同拆方对TLR-4/MyD88/NF-κB通路的影响。方法:64只Wistar大鼠随机分为正常对照组8只、模型组56只,采用中医证候联合乙醇-2,4,6-三硝基苯磺酸灌肠法复制UC大鼠模型,造模成功后,将模型大鼠随机分为模型组(M)、美沙拉嗪组(A)、健脾化滞丸全方组(B)、健脾清化方组(C)、健脾清化活血方组(D)、健脾清化导滞方组(E)、清化导滞活血方组(F),每组8只,给予相应药物灌胃4周,疗程结束后处死所有大鼠并取材,观察大鼠体质量变化、疾病活动度及结肠病理改变,ELISA及免疫组化检测大鼠结肠组织TNF-α表达;RT-PCR及Western blot检测大鼠结肠组织TLR-4、MyD88、NF-κB mRNA及蛋白表达。结果:健脾化滞丸全方、健脾清化方、健脾清化活血方组及健脾清化导滞方组大鼠体质量明显高于模型组(P<0.01),各治疗组均能明显改善UC模型大鼠结肠组织病理损伤及疾病活动指数,其中健脾化滞丸全方组及健脾清化活血方组作用最好。各治疗组均可抑制结肠组织TLR-4、MyD88、NF-κB mRNA及蛋白表达(P<0.001),其中美沙拉嗪组、健脾化滞丸全方组、健脾清化导滞方组及清化导滞活血方组在降低NF-κB mRNA及蛋白表达上明显优于健脾清化方组及健脾清化活血方组(P<0.05)。各组均能降低结肠组织TNF-α表达(P<0.001),组间差异无统计学意义(P>0.05)。结论:健脾化滞丸及各拆方均可能通过TLR-4/MyD88/NF-κB通路发挥作用,其中黄连、煨木香、凤尾草、炮姜可能为本方核心药物,对临床具有一定指导意义。

水飞蓟素通过共同抑制TLR4/NF-κB和TNF-α/ROS/P38MAPK通路减轻糖尿病肾病大鼠肾脏损伤的研究

目的:探讨水飞蓟素通过共同抑制Toll样受体4/核因子-κB(TLR4/NF-κB)和肿瘤坏死因子α/活性氧簇/P38丝裂原活化蛋白激酶(TNF-α/ROS/P38MAPK)通路减轻糖尿病肾病(DN)大鼠肾脏损伤的机制。方法:选取健康SD大鼠50只,按随机数字表法分为对照组、模型组、低剂量水飞蓟素组、中剂量水飞蓟素组、高剂量水飞蓟素组,每组10只。建模成功并治疗8周后,生化分析仪测定各组大鼠空腹血糖(FBG)、血肌酐(Scr)、尿素氮(BUN)、胱抑素C(Cys-C)、β_(2)微球蛋白(β_(2)-MG)、24 h尿蛋白(UTP)水平;计算肾脏指数;试剂盒检测各组大鼠肾组织丙二醛(MDA)、总抗氧化能力(T-AOC)水平;RT-PCR检测各组大鼠TLR4,NF-κB,TNF-α和P38MAPK等mRNA的表达水平。结果:与对照组相比,模型组体质量、T-AOC均降低(P<0.05),肾脏指数和FBG,Scr,BUN,Hcy,Cys-C,β_(2)-MG,24 h UTP,MDA,TLR4 mRNA,NF-κB p65 mRNA,TNF-αmRNA,P38MAPK mRNA均升高(P<0.05)。低、中、高剂量水飞蓟素组体质量、T-AOC水平均低于对照组,且均高于模型组(P<0.05);低、中、高剂量水飞蓟素组肾脏指数和FBG,Hcy,Cys-C,β_(2)-MG,24 h UTP,MDA,TLR4 mRNA,NF-κB p65 mRNA,TNF-αmRNA,P38MAPK mRNA均高于对照组,且均低于模型组(P<0.05);低、中剂量水飞蓟素组Scr,BUN水平均高于对照组,且低于模型组(P<0.05);高剂量水飞蓟素组Scr,BUN水平均低于模型组(P<0.05),高于对照组,但差异无统计学意义(P>0.05)。结论:水飞蓟素可调控TLR4/NF-κB和TNF-α/ROS/P38MAPK通路,可通过抑制其激活减轻氧化应激、减轻DN大鼠的肾脏损伤。

NF-κB在脂多糖诱导小鼠脓毒症急性肺损伤中对ANXA2表达的影响

目的观察脂多糖(LPS)诱导脓毒症急性肺损伤(ALI)小鼠肺组织中膜联蛋白A2(ANXA2)的表达水平,探讨ANXA2的表达与核转录因子-κB(NF-κB)信号通路的关系。方法健康雄性C57BL/6小鼠18只,随机分为空白对照组、脂多糖组(LPS组)及LPS+NF-κB抑制剂组(以下为抑制剂组),每组6只。抑制剂组腹腔注射NF-κB信号通路抑制剂吡咯烷二硫代甲酸铵(PDTC)50 mg/kg,1 h后LPS组及抑制剂组腹腔注射LPS 7.5 mg/kg,观察12 h,全身麻醉后采血并处死小鼠。HE染色观察肺组织病理学特征,测右肺中叶湿干重、计算湿/干重比值,ELISA法检测血清ANXA2及左肺肺泡灌洗液中ANXA2、肿瘤坏死因子-α(TNF-α)水平,免疫组化法检测肺组织ANXA2、NF-κBp65、p-NF-κBp65蛋白平均光密度(MOD)值,RT-qPCR检测肺组织中NF-κBp65、TNF-α及IL-6 mRNA相对表达量,WB检测肺组织ANXA2、NF-κBp65、p-NF-κBp65、Claudin1蛋白表达量。结果与空白对照组比较,LPS组肺组织病理损伤严重,可见细支气管上皮细胞坏死脱落、肺泡间隔增厚、肺泡腔内巨噬细胞和淋巴细胞渗出、少许出血。与空白对照组比较,LPS组右肺中叶湿/干重比值,血清ANXA2及肺泡灌洗液ANXA2、TNF-α水平,肺组织NF-κBp65、TNF-α及IL-6 mRNA相对表达量,肺组织ANXA2、NF-κBp65、p-NF-κBp65蛋白MOD值以及蛋白表达均显著升高(P<0.05),而Claudin1蛋白表达显著下降(P<0.05);与LPS组比较,抑制剂组以上各项观察指标均明显改善(P<0.05)。结论NF-κB信号通路在LPS诱导小鼠脓毒症相关ALI肺组织中可调控ANXA2的表达。

参附注射液对创伤失血性休克肝损伤大鼠NF-κB、TNF-α表达的影响

[目的]观察大鼠创伤失血性休克肝损伤后核转录因子-κB(NF-κB)、肿瘤坏死因子-α(TNF-α)表达的情况及不同剂量参附注射液对NF-κB、TNF-α表达的调控作用,探讨参附注射液对创伤失血性休克肝损伤的保护机制。[方法]取100只Wistar雄性大鼠随机分为空白对照组、模型对照组和参附注射液低、中、高剂量组,每组20只。除空白对照组外各组大鼠做双侧股骨骨折合并休克实验制造模型,各组分别于造模后0 h、2 h、4 h、6 h进行药物干预,空白对照组和模型对照组予生理盐水腹腔注射,参附注射液低、中、高剂量组分别以5 ml/kg、10 ml/kg、20 ml/kg剂量腹腔注射给药。实验过程中死亡大鼠立即进行解剖,切取肝脏标本。空白对照组大鼠直接活杀取相应组织及其标本。采用免疫组化法检测各组大鼠肝脏的NF-κB和TNF-α的蛋白表达水平。[结果]大鼠创伤失血性休克后NF-κB和TNF-α在肝组织中迅速表达,伤后6 h左右达到含量的高峰,伤后8 h仍持续在相当高的水平。与空白对照组比较,模型对照组及参附注射液低、中、高剂量组不同时间点(0.5 h、2 h、4 h、6 h、8 h)肝组织NF-κB、TNF-α表达水平均明显升高,差异有统计学意义(P<0.05);与模型对照组比较,参附注射液低、中、高剂量组不同时间点肝组织NF-κB、TNF-α表达水平均有不同程度下降,差异有统计学意义(P<0.05);参附注射液低、中、高剂量组比较,不同时间点肝组织NF-κB、TNF-α的下降水平与剂量呈正相关关系,差异有统计学意义(P<0.05)。[结论]参附注射液通过抑制NF-κB和TNF-α的表达而减少创伤失血性休克的损伤,降低死亡率,进而发挥对肝脏的保护作用。

小檗碱激活AMPK/NF-κB/TNF-α信号通路对肾病综合征大鼠肾损伤的影响

目的:从5’-单磷酸腺苷活化蛋白激酶(AMPK)/核转录因子-κB(NF-κB)/肿瘤坏死因子-α(TNF-α)信号通路,探究小檗碱改善肾病综合征(NS)大鼠肾损伤的可能机制。方法:尾静脉注射阿霉素建立NS大鼠模型,随机分为模型组、小檗碱治疗组、激素治疗组、AMPK抑制剂组、小檗碱+AMPK抑制剂组,每组15只,另取15只对照组大鼠。各组连续给药治疗5周后,收集尿液检测24 h尿蛋白;取动脉血检测血浆白蛋白、血脂水平;PAS染色观察肾组织形态变化;Western blot法检测肾组织AMPK、p-AMPK、NF-κB、p-NF-κB、TNF-α、白细胞介素-6(IL-6)、脂肪酸移位酶(FAT/CD36)、P糖蛋白(P-gp)表达水平。结果:与对照组相比,模型组大鼠肾小球血管袢与球囊壁黏连、肾小管管腔狭窄、肾小管上皮细胞水肿、坏死及空泡化等病理损伤严重,24 h尿蛋白、血脂水平升高,血浆白蛋白含量、AMPK活性、与AMPK有关的炎性反应蛋白及促脂质吸收相关蛋白表达降低(P<0.05)。小檗碱治疗或激素治疗,均可改善NS大鼠肾脏损伤,降低24 h尿蛋白、血脂水平,升高血浆中白蛋白含量,促进AMPK活化,发挥抗炎及改善脂质代谢作用(P<0.05),激素治疗组可引起P-gp表达升高,且其对NS大鼠的治疗效果弱于小檗碱治疗组(P<0.05)。AMPK抑制剂可加重NS大鼠肾损伤,并削弱小檗碱促进AMPK活化、改善NS大鼠肾损伤等作用(P<0.05)。结论:小檗碱可通过促进AMPK活化,改善NS大鼠糖脂代谢、抑制NF-κB/TNF-α炎症通路,发挥肾保护作用。

IL-6对种植体周围炎大鼠牙龈组织中TNF-α表达的影响

目的 探究白细介素(IL-6)对种植体周围炎大鼠牙龈组织中肿瘤坏死因子-α(TNF-α)表达的机制。方法 按照随机数字表法将40只大鼠分为假手术组(sham组)、种植体周围炎大鼠模型组(PI组)、种植体周围炎大鼠模型给予IL-6抑制剂Tocilizumab组(Tocilizumab组)、在Tocilizumab组的基础上给予NF-κB/p65激活剂LPS组(LPS组),每组10只。观察种植体周围软组织袋深度(PPD)、探诊出血(BOP)、出血指数(BI)和牙龈指数(GI)。测量种植体周围骨高度和骨密度(Micro-CT法);检测种植体周围牙龈组织炎症浸润(HE染色);RT-PCR检测种植体周围牙龈组织中IL-6、TNF-α mRNA表达;检测牙龈组织中p-p65蛋白表达(蛋白质印迹法)。结果 与sham组比较,PI组大鼠种植体软组织中PPD(近颊、远颊、近舌、远舌)、BOP、BI和GI均明显增加(P<0.05);与PI组比较,Tocilizumab组大鼠种植体软组织中PPD(近颊、远颊、近舌、远舌)、BOP、BI和GI均明显降低(P<0.05);与Tocilizumab组比较,种植体软组织中PPD(近颊、远颊、近舌、远舌)、BOP、BI和GI均明显增加(P<0.05)。与sham组比较,PI组大鼠种植体骨高度、牙槽骨密度明显降低,IL-6、TNF-α mRNA和p-p65蛋白表达增加(P<0.05);与PI组比较,Tocilizumab组大鼠种植体骨高度、牙槽骨密度明显升高,IL-6、TNF-α mRNA和p-p65蛋白表达低于(P<0.05);LPS组大鼠种植体骨高度、牙槽骨密度低于Tocilizumab组,IL-6、TNF-α mRNA和p-p65蛋白表达高于Tocilizumab组(P<0.05)。结论 IL-6抑制剂可通过抑制牙髓组织中NF-κB通路激活,进而抑制种植体周围炎大鼠牙龈组织中TNF-α释放,从而减轻大鼠种植体周围炎发展。

基于NF-κB探讨启宫丸对PCOS-IR大鼠炎症的影响

目的:研究启宫丸对PCOS-IR大鼠血清TNF-α、IL-6及子宫组织NF-κB的影响,探讨启宫丸改善PCOS慢性炎症的可能机制。方法:60只SPF级雌性SD大鼠按随机数字表法随机分为正常对照组、模型组、二甲双胍(270 mg/kg)组及启宫丸低(3.78 g/kg)、中(7.56 g/kg)、高(15.12 g/kg)剂量组。采用来曲唑联合高脂饮食建立PCOS-IR模型。造模后各给药组大鼠分别灌胃相应药物,正常对照组和模型组大鼠灌胃相应体积生理盐水,连续21 d。ELISA检测大鼠血清TNF-α、IL-6水平;免疫组化法和Western Blot法检测大鼠子宫组织中NF-κBp65蛋白表达;Real-time PCR检测大鼠子宫组织中NF-κB mRNA表达。结果:与正常对照组比较,模型组大鼠处在动情间期;子宫内膜腺体少、短而直;血清TNF-α、IL-6和子宫NF-κB mRNA及NF-κBp65蛋白表达显著升高(P<0.05或P<0.01)。与模型组比较,启宫丸中、高剂量组大鼠子宫内膜腺体多且弯曲;血清TNF-α、IL-6和子宫NF-κBp65蛋白表达显著降低,启宫丸高剂量组大鼠子宫NF-κB mRNA表达显著降低(P<0.05或P<0.01)。结论:启宫丸能改善PCOS大鼠慢性炎症,给子宫内膜的发育提供一个良好的微环境,其机制可能与NF-κB通路有关。

Quercetin regulates depression-like behavior in CUMS rat models via TLR4/NF-κB signaling

Background:Depression is becoming increasingly prevalent around the world,imposing a substantial burden on individuals,families,as well as society.Quercetin is known to be highly effective in treating depression.However,additional research is needed to dissect the mechanisms of its anti-depressive effects.Methods:For this study,Sprague-Dawley(SD)rats were randomized into the control,model,quercetin,or fluoxetine group.The latter three groups were exposed to chronic unpredictable mild stress(CUMS)for 42 d.The first two groups received saline solution daily via oral gavage.Meanwhile,the quercetin group was orally administered a quercetin suspension(52.08 mg/kg)every day,while the fluoxetine group was orally administered a fluoxetine solution(2.08 mg/kg).Here,fluoxetine served as the positive control drug to compare the therapeutic effects of quercetin.The experimental period was 6 weeks.Depressive behaviors in rats were assessed through various physiological and behavioral measures.Additionally,pathological changes in hippocampal tissues were examined using Nissl staining.Serum cytokines were detected using an enzymelinked immunosorbent assay(ELISA),and immunohistochemistry was employed to quantify the levels and integral optical density(IOD)values of ionized calcium binding adaptor molecule-1(Iba-1)expression in the brain.Real-time fluorescence quantitative PCR(RT-qPCR)was utilized to evaluate the mRNA levels of inflammatory indicators as well as toll-like receptor 4(TLR4),and nuclear factor-κappa B P65(NF-κB P65)in hippocampus.Western blot(WB)technique was employed to observe the protein levels of TLR4,NF-κB P65,and phospho-NF-κB P65(p-NF-κB P65).Results:After 42 d of exposure to CUMS,rats exhibited a slow increase in body weight,a reduction in food intake,an abnormal preference for sugar water,and aberrant open-field behaviors.Pathological analysis revealed the disintegration,rupture,interruption,and disorganization of hippocampal neuronal cells after CUMS exposure,along with a decrease in Nissl bodies in the CA1 region.This was accompanied by the elevated expression of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6)in the serum and the upregulation of IL-1β,IL-6,and TNF-αmRNA expression in the hippocampus.Increases in Iba-1-positive cells and the IOD values of Iba-1 were detected in hippocampal microglia.Furthermore,TLR4 and NF-κB P65 mRNA and protein levels were upregulated in hippocampal tissues.Quercetin,an antidepressant,could alleviate depression-like symptoms in rats and downregulate inflammatory factors associated with the TLR4/NF-κB signaling pathway in hippocampal microglia,and its therapeutic effect was comparable to fluoxetine.Conclusion:In rat models of CUMS,quercetin may act as an antidepressant by inhibiting inflammation in hippocampal microglia via TLR4/NF-κB signaling pathway.These results offer experimental and theoretical support for applying quercetin in the clinical management of depression.

基于NF-κB/TNF-α/IL-6通路探讨拟黑多刺蚁活性组分对脂多糖诱导抑郁小鼠神经炎症的影响

目的:探讨拟黑多刺蚁活性组分对脂多糖(LPS)所诱导抑郁小鼠神经炎症的影响及其可能的作用机制。方法:将60只雄性C57BL/6J小鼠随机分为正常对照组、模型组、氟西汀(20 mg/kg)阳性对照组及拟黑多刺蚁活性组分高(8 g/kg)、中(4 g/kg)、低(2 g/kg)剂量组,连续给药10 d,第11、12天采用LPS(1 mg/kg)腹腔注射诱导小鼠抑郁模型。对小鼠进行行为学实验(强迫游泳实验、悬尾实验),ELISA法检测小鼠血清神经递质和炎症因子,HE、Tunel染色观察小鼠脑组织病理损伤和凋亡情况,Real-time PCR检测小鼠脑组织中IL-6 mRNA表达,Western Blot检测小鼠脑组织中NF-κB/TNF-α/IL-6通路相关蛋白表达。将RAW264.7细胞分为空白对照组、模型组及拟黑多刺蚁活性组分高(2 mg/mL)、中(1 mg/mL)、低(0.5 mg/mL)浓度组,加入脂多糖(1μg/mL)造模,Western Blot检测各组细胞中NF-κB/TNF-α/IL-6通路相关蛋白表达。结果:动物实验中,与正常对照组比较,模型组小鼠的游泳不动时间、悬尾不动时间、血清TNF-α水平及脑组织IL-6 mRNA和p-NF-κB、IL-6、TNF-α蛋白表达显著升高(P<0.05或P<0.01),脑组织病理变化明显,脑组织神经元凋亡增加。与模型组比较,除拟黑多刺蚁活性组分低剂量组小鼠血清TNF-α水平及中剂量组小鼠血清TNF-α、DA、BDNF水平差异无统计学意义(P>0.05)外,拟黑多刺蚁活性组分各组小鼠游泳不动时间、悬尾不动时间、血清TNF-α水平、脑组织IL-6 mRNA表达及脑组织IL-6、TNF-α、p-NF-κB蛋白表达显著降低,血清5-HT、DA、BDNF水平显著升高(P<0.05或P<0.01),脑组织病理学明显改善,脑组织神经元凋亡明显减少。体外实验中,与空白对照组比较,模型组RAW264.7细胞IL-6、TNF-α、p-NF-κB蛋白表达显著升高(P<0.05或P<0.01),与模型组比较,拟黑多刺蚁活性组分各组RAW264.7细胞TNF-α、p-NF-κB蛋白表达显著降低,高浓度组IL-6蛋白表达显著降低(P<0.05或P<0.01)。结论:拟黑多刺蚁活性组分可通过抑制NF-κB/TNF-α/IL-6通路的激活,改善小鼠抑郁症神经炎症。

舒肝和络醒脾方对肝纤维化模型大鼠TNF-α/NF-κB信号通路的影响

目的:观察舒肝和络醒脾方对肝纤维化模型大鼠TNF-α/NF-κB信号通路以及炎症因子的影响,探讨舒肝和络醒脾方干预肝纤维化进程的相关机制。方法:将雄性Wistar大鼠随机分为正常对照组,模型组,阳性对照组,舒肝和络醒脾方高、中、低剂量组共6组,每组10只。除正常对照组外,其余各组大鼠均采用腹腔注射40%CCl4油溶液方式诱导肝纤维化模型,注射同时采用相应药物进行灌胃治疗,8周后取大鼠肝脏与血液标本。造模期间观察记录各组大鼠一般情况;HE染色与Masson染色观察大鼠肝组织病理形态学变化;免疫组织化学法检测肝组织α-SMA表达水平;双抗体夹心ELISA法检测大鼠血清IL-1β、TNF-α炎症因子水平;RT-q PCR法检测TNF-α、NF-κB p65的m RNA表达水平;Western blot技术检测TNF-α、NF-κB p65的蛋白表达水平。结果:同模型组比较,大鼠经药物干预后一般状况均改善;肝脏病理形态明显改善(P<0.05);α-SMA表达水平明显下降(P<0.05);炎症因子IL-1β与TNF-α表达水平明显下降(P<0.05);TNF-α、NF-κB p65的m RNA水平和蛋白表达均明显下降(P<0.05)。其中舒肝和络醒脾方高剂量组效果较为显著且稳定,和阳性对照组疗效趋同。结论:舒肝和络醒脾方可延缓由CCl4诱导的肝纤维化进程,其作用机制可能与TNF-α/NF-κB信号通路有关。

Huangqin decoction alleviates lipid metabolism disorders and insulin resistance in nonalcoholic fatty liver disease by triggering Sirt1/NF-κB pathway

BACKGROUND Nonalcoholic fatty liver disease(NAFLD)is a clinicopathological entity characterized by intrahepatic ectopic steatosis.As a consequence of increased consumption of high-calorie diet and adoption of a sedentary lifestyle,the incidence of NAFLD has surpassed that of viral hepatitis,making it the most common cause of chronic liver disease globally.Huangqin decoction(HQD),a Chinese medicinal formulation that has been used clinically for thousands of years,has beneficial outcomes in patients with liver diseases,including NAFLD.However,the role and mechanism of action of HQD in lipid metabolism disorders and insulin resistance in NAFLD remain poorly understood.AIM To evaluate the ameliorative effects of HQD in NAFLD,with a focus on lipid metabolism and insulin resistance,and to elucidate the underlying mechanism of action.METHODS High-fat diet-induced NAFLD rats and palmitic acid(PA)-stimulated HepG2 cells were used to investigate the effects of HQD and identify its potential mechanism of action.Phytochemicals in HQD were analyzed by highperformance liquid chromatography(HPLC)to identify the key components.RESULTS Ten primary chemical components of HQD were identified by HPLC analysis.In vivo,HQD effectively prevented rats from gaining body and liver weight,improved the liver index,ameliorated hepatic histological aberrations,decreased transaminase and lipid profile disorders,and reduced the levels of pro-inflammatory factors and insulin resistance.In vitro studies revealed that HQD effectively alleviated PA-induced lipid accumulation,inflammation,and insulin resistance in HepG2 cells.In-depth investigation revealed that HQD triggers Sirt1/NF-κB pathwaymodulated lipogenesis and inflammation,contributing to its beneficial actions,which was further corroborated by the addition of the Sirt1 antagonist EX-527 that compromised the favorable effects of HQD.CONCLUSION In summary,our study confirmed that HQD mitigates lipid metabolism disorders and insulin resistance in NAFLD by triggering the Sirt1/NF-κB pathway.