姜黄素对亚急性肝内胆汁淤积大鼠肠内菌群变化的影响及其机制研究

目的： 研究姜黄素对亚急性肝内胆汁淤积（intrahepatic cholestasis,IHC）大鼠肠内菌群变化的影响,并分析相关机制。方法： 38只IHC大鼠随机分为模型组（9只）、姜黄素低剂量组（9只）、姜黄素高剂量组（10只）、阳性药物组（10只）,9只健康大鼠设置为对照组。姜黄素低、高剂量组姜黄素200、400 mg·kg^-1灌胃;阳性药物组熊去氧胆酸60 mg·kg^-1灌胃;对照组与疾病组等量生理盐水灌胃。均为每天一次;干预14 d。检测肝功能指标、胆红素代谢指标、炎症指标;qRT-PCR反应定量检测肠内菌群;Western blot检测肝组织核转录因子-κB（NF-κB）p65、p-NF-κB p65、核因子抑制蛋白（IκBα）、p-IκBα蛋白相对表达量。结果： 与疾病组比较,姜黄素低、高剂量组、阳性药物组碱性磷酸酶（ALP）、丙氨酸氨基转移酶（ALT）、谷草转氨酶（AST）、γ-谷氨酰转肽酶（GGT）、总胆红素（TBIL）、直接胆红素（DBIL）、总胆汁酸（TBA）、白介素-1β（IL-1β）、白介素-6（IL-6）、转化生长因子-β₁（TGF-β₁）均降低,白介素-10（IL-10）均升高,乳酸杆菌、双歧杆菌均增加,大肠杆菌均减少（P<0.05）;与姜黄素低剂量组比较,姜黄素高剂量组、阳性药物组ALP、ALT、AST、GGT、TBIL、DBIL、TBA、IL-1β、IL-6、TGF-β₁均降低,IL-10均升高,乳酸杆菌、双歧杆菌均增加,大肠杆菌均减少（P<0.05）。与疾病组比较,姜黄素低、高剂量组、阳性药物组p-NF-κB p65/NF-κB p65,p-IκBα/IκBα均降低（P<0.05）;与姜黄素低剂量组比较,姜黄素高剂量组、阳性药物组p-NF-κB p65/NF-κB p65,p-IκBα/IκBα均降低（P<0.05）。结论： 姜黄素可改善亚急性IHC模型大鼠肝功能、胆红素代谢、肠道菌群,减轻炎症反应及肝脏组织病理变化,且具有剂量依赖性,推测其作用机制与抑制NF-κB信号通路有关。     

姜黄素促进损伤骨骼肌修复

目的:研究姜黄素促进损伤骨骼肌修复的功能.方法:制作小鼠胫骨前肌实验性损伤模型,实验小鼠分别肌注和口服姜黄素,以组织学指标观察姜黄素体内促进损伤骨骼肌的修复功能.采用MTT和流析细胞分析技术观察姜黄素对骨骼肌成肌细胞的作用及可能的信号传导通路.结果:实验的小鼠肌肉伤口区新生肌纤维显著多于对照组.姜黄素无明显促C2C12细胞增殖,但能促进C2C12细胞融合而分化成肌纤维;姜黄素能抑制C2C12细胞NFκB反应因子的转活,而对c-Myc和CRE通路反应因子没有显著影响.大剂量使用姜黄素未见急性毒性反应.结论:姜黄素能通过促进骨骼肌干细胞的分化而促进损伤骨骼肌的修复.     

甲氨蝶呤对脂多糖诱导的大鼠脊髓神经胶质细胞pIκBα-NF-κBp65-炎性因子通路的影响

目的 观察甲氨蝶呤对脂多糖（lipopolysaccharide,LPS）诱导的大鼠脊髓神经胶质细胞pIκBα-NF-κBp65-炎性因子通路的影响。方法 脊髓组织块法培养神经胶质细胞。将分离的神经胶质细胞接种于多孔板培养48 h后,分为空白对照组、LPS组、LPS+pIκBα抑制剂组、LPS+甲氨喋呤组。随后应用免疫印迹法测定各组分的pIκBα与胞核及胞浆NF-κBp65水平变化,酶免疫法（ELISA）测定炎性因子TNF-α、IL^-1β、IL-6含量。结果 神经胶质细胞经LPS诱导后,pIκBα、胞核NF-κBp65和细胞上清液炎性因子TNF-α、IL^-1β、IL-6水平均显著增加（P<0.05或P<0.01）。甲氨喋呤可明显抑制经LPS诱导的神经胶质细胞pIκBα水平,显著降低胞核NF-κBp65水平和细胞上清液炎性因子TNF-α、IL^-1β、IL-6的含量（P<0.05）。结论 甲氨喋呤对LPS诱导的脊髓神经胶质细胞pIκBα-NF-κBp65-炎性因子通路有显著的抑制作用。     

金合欢素对糖尿病肾病大鼠肾损伤及TLR4/NF-κB信号通路的影响

目的 探讨金合欢素对糖尿病肾病（DN）大鼠肾损伤及Toll样受体4（TLR4）/核因子-κB（NF-κB）信号通路的影响。方法 通过高脂高糖饲料喂养及ip链脲佐菌素（STZ）建立DN大鼠模型,将造模成功的大鼠随机分为模型组,金合欢素低、高剂量（40、80 mg ·kg^-1）组,金合欢素（80 mg ·kg^-1）＋脂多糖（LPS,0.1 mg ·kg^-1）组,缬沙坦（20 mg ·kg^-1）组,每组10只,并以正常喂养且不ip STZ的10只大鼠作为对照组。干预结束后,尾静脉取血,检测大鼠空腹血糖含量;收集大鼠24 h尿液,分析尿蛋白含量;腹部主动脉取血,ELISA法检测血清中血肌酐、血尿素氮水平及肿瘤坏死因子-α （TNF-α）、白细胞介素-6（IL-6）水平;分离双肾组织,透射电镜及HE染色检测肾组织损伤情况;Western blotting检测Toll样受体4（TLR4）/核因子-κB（NF-κB）通路相关蛋白表达。结果 与对照组比较,模型组肾组织严重受损,血糖、尿蛋白、血肌酐、血尿素氮、血清TNF-α和IL-6水平、肾组织TLR4和p-NF-κB p65/NF-κB p65蛋白表达显著增加（P<0.05）;与模型组比较,缬沙坦和金合欢素低、高剂量组肾组织损伤得到缓解,血糖、尿蛋白、血肌酐、血尿素氮、血清TNF-α和IL-6水平、肾组织TLR4和p-NF-κB p65/NF-κB p65蛋白表达显著降低（P<0.05）;与金合欢素高剂量组相比,金合欢素＋LPS组肾组织损伤加剧,血糖、尿蛋白、血肌酐、血尿素氮、血清TNF-α和IL-6水平、肾组织TLR4和p-NF-κB p65/NF-κB p65蛋白表达显著增加（P<0.05）。结论 金合欢素通过抑制TLR4/NF-κB信号通路改善DN大鼠肾损伤。     

姜黄素调控NF-κB信号通路对糖尿病模型大鼠胰岛细胞形态和功能的影响

目的 探讨姜黄素通过核因子-κB（NF-κB）信号通路对高糖高脂饮食诱导的糖尿病模型大鼠胰岛细胞形态和功能的影响。方法 采用高糖高脂饲料＋ip链脲佐菌素法制备糖尿病大鼠模型,将模型成功 SD大鼠随机分为5组：模型组、二甲双胍（200 mg·kg^-1,阳性药）组和姜黄素低、中、高剂量（50、150、250 mg·kg^-1）组,对照组不造模。ig给药,每天1次,连续6周,对照组同时注射等量无菌磷酸盐缓冲液（PBS）。观察SD大鼠体质量变化;血糖仪检测空腹血糖;ELISA法检测血清C肽水平;生化分析仪检测血清中糖化血红蛋白、三酰甘油（TG）、总胆固醇（TC）、低密度脂蛋白胆固醇（LDL-C）、高密度脂蛋白胆固醇（HDL-C）、丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、总胆红素（TBil）、血肌酐（Scr）、血尿素氮（BUN）、尿酸（UA）水平;取胰腺进行 HE 染色,于光镜下观察胰岛形态学改变;免疫组化染色法观察胰岛 NF-κB 的阳性表达;Western blotting法检测NF-κB信号通路相关蛋白表达;提取胰岛细胞,免疫荧光染色观察胰岛素分泌,葡萄糖刺激的胰岛素分泌（GSIS）实验检测胰岛素分泌量。结果 与模型组比较,各给药组体质量在治疗第2周时开始缓慢增长,第3、4周体质量显著升高（P＜0.05）;血糖水平显著降低（P＜0.05）,血清 C 肽水平显著升高（P＜0.05）;糖化血红蛋白、TG、TC、LDL-C、ALT、AST、Scr、BUN、UA水平显著降低（P＜0.05）,HDL-C水平显著升高（P＜0.05）;胰岛大小和形态逐渐恢复正常,胰岛细胞依次变得更清晰完整,导管系统内的囊性扩张也逐渐减少;NF-κB阳性细胞减少;p-P65蛋白表达显著降低（P＜0.05）,p-IκBα蛋白表达显著升高（P＜0.05）;胰岛原代细胞分泌胰岛素显著增加（P＜0.05）。结论 姜黄素可以保护糖尿病大鼠胰岛细胞正常形态,维持胰岛细胞功能,减轻糖尿病大鼠症状,其机制可能与调控NF-κB信号通路有关。     

PI3K/Akt/mTOR通路在炎症相关疾病中分子机制研究进展

炎症是一种机体应对感染、组织损伤或者细胞应激的反应,并且可以通过修复机制恢复组织功能。炎症发生时会引起多条信号通路的激活,包括核转录因子-κB（NF-κB）通路、Janus激酶/信号转导与转录激活子（JAK/STAT）通路、丝裂原活化蛋白激酶（MAPK）通路以及磷脂酰肌醇-3-激酶/蛋白激酶B /雷帕霉素靶蛋白（PI3K/Akt/mTOR）通路等。本文综述了近年来磷脂酰肌醇-3-激酶/蛋白激酶B /雷帕霉素靶蛋白通路在炎症相关疾病中的分子作用机制,为研发以磷脂酰肌醇-3-激酶/蛋白激酶B /雷帕霉素靶蛋白为靶点的药物提供理论依据。     

姜黄素下调NF-κB信号通路抑制化学缺氧诱导的U87细胞炎症反应

目的 研究姜黄素对化学缺氧所致人源性神经星形胶质瘤细胞系U87炎症反应的影响,并探讨相关分子机制。方法 100 μmol/L CoCl₂处理U87细胞不同时间（1、3、6、12、24 h）,实时荧光定量PCR（qRT-PCR）法检测炎症因子白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α） mRNA表达变化。100 μmol/L CoCl₂处理U87细胞12 h制备化学缺氧模型,同时给予1、5和10 μmol/L姜黄素处理,对照组（不添加CoCl₂）和模型组给予等体积DMSO;qRTPCR法检测IL-1β、IL-6、TNF-α mRNA表达变化;Western Blotting法检测NF-κB/P65蛋白磷酸化水平及核转位。结果 CoCl₂上调U87细胞IL-1β、IL-6、TNF-α mRNA水平并呈时间相关性,炎症反应在约12 h达到高峰。与模型组比较,姜黄素显著下调炎症因子IL-1β、IL-6和TNF-α的mRNA水平,5和10 μmol/L浓度组差异显著（P<0.05）;显著下调p65的磷酸化水平（P<0.05）;导致细胞核内NF-κB/p65蛋白显著减少、细胞质中NF-κB/p65蛋白显著增加（P<0.05）。结论 姜黄素通过下调NF-κB信号通路抑制化学缺氧诱导的U87细胞炎症反应。     

绿原酸抑制NF-κB/NLRP3炎性体通路减轻LPS诱导小胶质细胞神经炎症损伤研究

摘要：目的:探究绿原酸（CGA）对脂多糖（LPS）诱导的小胶质细胞神经炎症损伤及核因子κB（NF-κB）/Nod样受体家族含pyrin结构域蛋白3（NLRP3）炎性体通路的影响。方法:培养小胶质细胞BV2,3-（4,5-二甲基噻唑-2）-2,5-二苯基四氮唑溴盐（MTT）法检测不同浓度(0,1,2,4,8 mmol·L-1)CGA对BV2细胞活力的影响;再次培养BV2细胞,依次分为对照（Control）组、LPS组、脂多糖和药物溶剂对照[LPS+二甲亚砜（DMSO）]、脂多糖和阳性药物对照（LPS+Positive）组以及脂多糖和绿原酸处理（LPS+CGA）组。免疫荧光法检测肿瘤坏死因子α（TNF-α）和白细胞介素（IL）-1β表达情况;ELISA法检测TNF-α、IL-1β、IL-6、IL-10、IL-12和诱导型一氧化氮合成酶（iNOS）含量;免疫印迹分析NF-κB/NLRP3炎性体通路和凋亡相关蛋白水平;流式细胞仪检测细胞凋亡率。结果:与CGA 0 mmol·L-1比较,CGA 1,2,4 mmol·L-1对BV2细胞增殖活力的影响无统计学意义（P>0.05）,CGA 8 mmol·L-1显著降低BV2细胞增殖活力（P<0.05）。LPS诱导后细胞凋亡率升高,细胞上清液中TNF-α、IL-1β、IL-6、IL-12和iNOS分泌量增多,IL-10分泌量减少,细胞中NF-κB p65、磷酸化NF-κB抑制蛋白（p-IκBα）、NLRP3、含半胱氨酸的天冬氨酸蛋白水解酶1（Caspase-1）和B细胞淋巴瘤/白血病-2（Bcl-2）相关X蛋白（Bax）蛋白水平上调,细胞中Bcl-2蛋白水平下调（P<0.05）。添加CGA可明显改善LPS对BV2细胞的影响（P<0.05）。结论:CGA通过抑制NF-κB/NLRP3炎性体通路减轻LPS诱导的BV2细胞神经炎症损伤。     

姜黄素抑制HMGB1-NF-κB信号通路减轻脂多糖诱导新生大鼠急性肺损伤实验研究

目的 分析姜黄素抑制高迁移率族蛋白B1（HMGB1）-核转录因子κB（NF-κB）信号通路减轻脂多糖（LPS）诱导新生大鼠急性肺损伤（ALI）的作用。方法 将60只新生SD雄性大鼠随机分为对照组、模型组、地塞米松（阳性药,2 mg·kg^-1）组和姜黄素低、中、高剂量（1.5、3.0、6.0 mg·kg^-1）组,每组10只。除对照组外,所有大鼠采用腹膜内注射LPS （3 mg·kg^-1）建立ALI模型。注射LPS约6 h后开始ip给药,每天1次,连续7 d,模型组和对照组大鼠ip等体积的0.1% DMSO。通过血氧分压（PaO₂）和肺干湿质量比（W/D）评估新生大鼠肺水肿情况;HE染色检测各组大鼠肺组织损伤;ELISA法检测新生大鼠支气管肺泡灌洗液（BALF）氧化应激指标超氧化物歧化酶（SOD）、髓过氧化物酶（MPO）、丙二醛（MDA）和谷胱甘肽（GSH）水平,BALF中白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）和HMGB1水平;Western blotting法检测大鼠肺组织胞核NF-κB、胞浆NF-κB和磷酸化核因子κB抑制因子α（p-IκBα）蛋白表达。结果 与对照组相比,模型组新生大鼠肺泡腔有渗出、肺组织结构紊乱、细胞核固缩深染、伴随大量的炎性细胞浸润,病理评分显著升高（P<0.01）; PaO₂、BALF中SOD和GSH水平显著降低（P<0.01）;肺W/D,BALF中MPO和MDA水平,BALF中IL-6、TNF-α和HMGB1水平,NF-κB胞核/胞浆比例和胞浆p-IκBα蛋白表达水平显著升高（P<0.01）。与模型组相比,姜黄素高、中剂量组和地塞米松组大鼠肺组织病理损伤减轻,肺泡腔渗出、炎性细胞浸润明显减少,病理评分显著降低（P<0.05、0.01）; PaO₂、BALF中SOD和GSH水平显著升高（P<0.05、0.01）;肺W/D,BALF中MPO、MDA水平,BALF中IL-6、TNF-α和HMGB1水平,NF-κB胞核/胞浆比例和胞浆p-IκBα蛋白表达水平显著降低（P<0.05、0.01）。结论 姜黄素可以通过抑制HMGB1-NF-κB信号通路减轻LPS诱导的新生大鼠ALI。     

银星养脑片对血管性痴呆模型大鼠的记忆改善作用及机制研究

目的 初步探讨银星养脑片治疗血管性痴呆的药效及可能机制。方法 构建双侧颈动脉结扎的血管性痴呆大鼠模型，术后14 d分别给予银星养脑片0.07，0.135，0.27 g·kg^-1及吡拉西坦片0.4 g·kg^-1，持续灌胃14 d后进行水迷宫行为学测试，随后收集血清及动物脑组织，检测血清中炎症因子TNF-α、IL-1β含量，对脑组织进行HE染色及尼氏染色观察海马区病理结构变化。使用BV2细胞构建LPS刺激模型，给予银星养脑片0.5，1，2 mg·kg^-1不同剂量后检测细胞上清炎症相关因子NO、IL-6、TNF-α的释放量，收集细胞蛋白检测iNOS、NF-κB p65、p-NF-κB p65、IκB-α、p-IκB-α、NLRP3、caspase-1炎症相关蛋白表达变化，免疫荧光检测细胞iNOS、NLRP3的表达情况。结果 水迷宫结果提示血管性痴呆大鼠出现明显的记忆损伤，而银星养脑片给药组能有效改善记忆损伤情况，且还能降低血管性痴呆大鼠血清炎症水平；在BV2细胞上给予银星养脑片的干预能减少LPS刺激后的炎症因子释放，抑制NF-κB p65和IκB-α的磷酸化蛋白的增加，抑制NF-κB下游NLRP3/caspase-1通路的激活，有效地起到抗炎的作用。结论 银星养脑片能有效改善血管性痴呆大鼠的记忆损伤，有可能通过抑制炎症相关NF-κB/NLRP3/caspase-1通路起到抗炎的作用。     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

Lung disease and PKCs

The lung offers a rich opportunity for development of therapeutic strategies focused on isozymes of protein kinase C (PKCs). PKCs are important in many cellular responses in the lung, and existing therapies for pulmonary disorders are inadequate. The lung poses unique challenges as it interfaces with air and blood, contains a pulmonary and systemic circulation, and consists of many cell types. Key structures are bronchial and pulmonary vessels, branching airways, and distal air sacs defined by alveolar walls containing capillaries and interstitial space. The cellular composition of each vessel, airway, and alveolar wall is heterogeneous. Injurious environmental stimuli signal through PKCs and cause a variety of disorders. Edema formation and pulmonary hypertension (PHTN) result from derangements in endothelial, smooth muscle (SM), and/or adventitial fibroblast cell phenotype. Asthma, chronic obstructive pulmonary disease (COPD), and lung cancer are characterized by distinctive pathological changes in airway epithelial, SM, and mucous-generating cells. Acute and chronic pneumonitis and fibrosis occur in the alveolar space and interstitium with type 2 pneumocytes and interstitial fibroblasts/myofibroblasts playing a prominent role. At each site, inflammatory, immune, and vascular progenitor cells contribute to the injury and repair process. Many strategies have been used to investigate PKCs in lung injury. Isolated organ preparations and whole animal studies are powerful approaches especially when genetically engineered mice are used. More analysis of PKC isozymes in normal and diseased human lung tissue and cells is needed to complement this work. Since opposing or counter-regulatory effects of selected PKCs in the same cell or tissue have been found, it may be desirable to target more than one PKC isozyme and potentially in different directions. Because multiple signaling pathways contribute to the key cellular responses important in lung biology, therapeutic strategies targeting PKCs may be more effective if combined with inhibitors of other pathways for additive or synergistic effect. Mechanisms that regulate PKC activity, including phosphorylation and interaction with isozyme-specific binding proteins, are also potential therapeutic targets. Key isotypes of PKC involved in lung pathophysiology are summarized and current and evolving therapeutic approaches to target them are identified.     

Gender-related symptoms and correlates of alcohol dependence among men and women with a lifetime diagnosis of alcohol use disorders

This study explored gender-related symptoms and correlates of alcohol dependence in a crosssectional study of 150 men and 150 women with a lifetime diagnosis of alcohol use disorders (AUD). Participants were recruited in equal numbers from treatment settings, correctional centres and the general community. Standardized measures were used to determine participants' use of substances, history of psychiatric disorders and psychosocial stress, their sensation seeking and family history of substance use and mental health disorders. Multivariate analyses were used to detect patterns of variables associated with gender and the lifetime severity of AUD. Men had a longer history of severe AUD than women. Women had similar levels of alcohol dependence and medical and psychological sequelae as men, despite 6 fewer years of AUD. More women than men had a history of severe psychosocial stress, severe dependence on other substances and antecedent mental health problems, especially mood and anxiety disorders. There were differences in family history of alcohol-related problems approximating same-gender aggregation. The severity of a lifetime AUD was predicted by its earlier age at onset and the occurrence of other disorders, especially anxiety, among both men and women. The limitations in the generalizability of these findings due to sample idiosyncrasies are discussed.     

Biochemical and molecular characterisation of cubozoan protein toxins

Class Cubozoa includes several species of box jellyfish that are harmful to humans. The venoms of box jellyfish are stored and discharged by nematocysts and contain a variety of bioactive proteins that are cytolytic, cytotoxic, inflammatory or lethal. Although cubozoan venoms generally share similar biological activities, the diverse range and severity of effects caused by different species indicate that their venoms vary in protein composition, activity and potency. To date, few individual venom proteins have been thoroughly characterised, however, accumulating evidence suggests that cubozoan jellyfish produce at least one group of homologous bioactive proteins that are labile, basic, haemolytic and similar in molecular mass (42-46 kDa). The novel box jellyfish toxins are also potentially lethal and the cause of cutaneous pain, inflammation and necrosis, similar to that observed in envenomed humans. Secondary structure analysis and remote protein homology predictions suggest that the box jellyfish toxins may act as α-pore-forming toxins. However, more research is required to elucidate their structures and investigate their mechanism(s) of action. The biological, biochemical and molecular characteristics of cubozoan venoms and their bioactive protein components are reviewed, with particular focus on cubozoan cytolysins and the newly emerging family of box jellyfish toxins.     

Dose tolerability of chronically inhaled voriconazole solution in rodents

Invasive pulmonary aspergillosis (IPA) is a fungal disease of the lung associated with high mortality rates in immunosuppressed patients despite treatment. Targeted drug delivery of aqueous voriconazole solutions has been shown in previous studies to produce high tissue and plasma drug concentrations as well as improved survival in a murine model of IPA. In the present study, rats were exposed to 20 min nebulizations of normal saline (control group) or aerosolized aqueous solutions of voriconazole at 15.625 mg (low dose group) or 31.25 mg (high dose group). Peak voriconazole concentrations in rat lung tissue and plasma after 3 days of twice daily dosing in the high dose group were 0.85 ± 0.63 μg/g wet lung weight and 0.58 ± 0.30 μg/mL, with low dose group lung and plasma concentrations of 0.38 ± 0.01 μg/g wet lung weight and 0.09 ± 0.06 μg/mL, respectively. Trough plasma concentrations were low but demonstrated some drug accumulation over 21 days of inhaled voriconazole administered twice daily. Following multiple inhaled doses, statistically significant but clinically irrelevant abnormalities in laboratory values were observed. Histopathology also revealed an increase in the number of alveolar macrophages but without inflammation or ulceration of the airway, interstitial changes, or edema. Inhaled voriconazole was well tolerated in a rat model of drug inhalation.