大柴胡颗粒对胆色素结石豚鼠保护作用机制研究

目的 观察大柴胡颗粒对胆色素结石豚鼠胆囊黏膜表皮生长因子(EGF)表达水平,肝、胆超微结构,肝组织胆固醇7α-羟化酶(CYP7Al)mRNA水平以及胆盐转运子BSEP、MRP2表达水平的影响,明确其对胆色素结石豚鼠的保护作用机制.方法 采用饲料法复制胆色素结行豚鼠模型,免疫组化法观察大柴胡颗粒(11、2.2、4.4 g/kg)对胆色素结石豚鼠的胆囊EGF水平的影响,透射电镜观察肝胆组织超微结构的改变,RT-PCR法检测肝组织CYP7A1基因表达水平,Westernblotting法检测肝脏中BSEP、MRP2表达水平,以熊去氧胆酸作为阳性对照.结果 大柴胡颗粒对胆结石豚鼠胆囊黏膜EGF表达影响不明显,但较好地改善其肝胆超微结构,其2.2、4.4 g/kg剂量组还能增加肝脏组织CYP7A1的基因转录(P＜0.05、0.01)和BSEP、MRP2蛋白表达水平(P＜0.05).结论 大柴胡颗粒抑制豚鼠胆色素结石形成可能与其影响豚鼠胆汁酸代谢、促进胆盐转运子功能、保护肝胆细胞器结构有关,而与胆囊黏膜EGF功能无明显关系.     

Nerolidol Protects Against LPS‐induced Acute Kidney Injury via Inhibiting TLR4/NF‐κB Signaling

Acute kidney injury (AKI) is a critical care syndrome, resulting in acute reduction of renal function and up to 22% mortality of hospitalized patients. Nerolidol is a major component in several essential oils that possesses various pharmacological properties. The present study aimed to investigate the potential effect of nerolidol on lipopolysaccharide (LPS)‐induced AKI. Nerolidol dose‐dependently reduced the pathological injuries of kidney induced by LPS in rats. Nerolidol significantly decreased the levels of blood urea nitrogen and creatinine in LPS‐treated rats in a dose‐dependent manner. In addition, nerolidol inhibited LPS‐induced decrease of cell viability in NRK‐52E rat proximal tubular cells, which effect was concentration dependent. Nerolidol notably inhibited the increase of TNFα and IL‐1β in LPS‐treated rats and the mRNA expression of TNFα and IL‐1β in LPS‐treated NRK‐52E cells. Nerolidol suppressed the increase of toll‐like receptor 4 (TLR4) expression, phosphorylation and nuclear translocation of p65 NF‐κB in kidneys of LPS‐treated rats and LPS‐treated NRK‐52E cells. Overexpression of TLR4 and p65 NF‐κB significantly suppressed nerolidol‐induced inhibition of TNFα and IL‐1β expression and increase of cell viability in LPS‐treated cells. In summary, we found that nerolidol played a critical anti‐inflammatory effects through inhibition of TLR4/NF‐κB signaling and protected against LPS‐induced AKI. Copyright © 2017 John Wiley & Sons, Ltd.     

芍药苷配伍小檗碱对 TNF-α诱导人脐静脉内皮细胞NF-κB/YY1 信号通路的影响

目的基于NF-κB/YY1信号通路探讨芍药苷配伍小檗碱对肿瘤坏死因子α(TNF-α)诱导的人脐静脉内皮细胞(HUVECs)炎症反应的影响。方法体外培养HUVECs,分别设空白对照组、模型组(TNF-α20 ng/m L)、芍药苷组(PF 160μmol/L)、小檗碱组(BBR 20μmol/L)、配伍组(160μmol/L芍药苷+20μmol/L小檗碱组)。采用CCK8法检测各组细胞活力;LDH毒性实验检测细胞上清液中LDH释放量;ELISA法检测细胞上清液中IL-6和IL-8含量;RT-PCR检测NF-κB、YY1基因表达;Western Blot法检测NF-κB、YY1蛋白表达。结果与空白对照组比较,模型组细胞活力减弱,LDH漏出率增加,IL-6和IL-8含量增加,NF-κB和YY1基因及蛋白表达上调(P<0.05,P<0.01)。与模型组比较,芍药苷组、小檗碱组及配伍组干预后细胞活力增强,LDH漏出率减少,IL6和IL-8含量降低,NF-κB和YY1基因及蛋白表达下调,且芍药苷和小檗碱配伍组较单体组干预后效果更明显(P<0.05,P<0.01)。结论TNF-α可激活HUVECs NF-κB/YY1信号通路,诱导炎症反应,加重内皮损伤。芍药苷配伍小檗碱较单药使用可更有效地抑制NF-κB/YY1信号通路,减轻炎症反应进而起到血管内皮保护作用。     

化脂复肝颗粒通过抑制TLR4/NF-κB/NLRP3减轻Kupffer细胞炎症改善非酒精性脂肪性肝病机制研究

目的:探讨化脂复肝颗粒在非酒精性脂肪性肝病(NAFLD)中对Kupffer细胞(KCs)TLR4/MyD88/NF-κB/NLRP3信号通路的影响。方法:将分离培养的KCs随机分为空白组、LPS干预组、LPS+空白血清组、LPS+化脂复肝组、高糖+对照组、高糖+LPS干预组、高糖+LPS+空白血清组、高糖+LPS+化脂复肝组,LPS干预浓度为100 ng/ml,高糖干预为于KCs培养基中另加入葡萄糖30 mmol/L。用CCK-8法检测各组KCs增殖情况; 酶联免疫吸附法(ELISA)测定各组KCs培养基上清液中促炎因子TNF-α、IL-1β的含量; 反转录实时荧光定量PCR(RT-qPCR)法检测各组KCs中TLR4、MyD88、NLRP3、Caspase-1、NF-κB p65、TNF-α、IL-1β的基因表达情况。结果:CCK-8检测显示,空白及高糖培养对照组KCs细胞不断增殖,长势良好; LPS+化脂复肝组与高糖+LPS+化脂复肝组KCs缓慢增殖,非高糖组KCs均较含高糖组KCs增殖速率更快; 而LPS或高糖+LPS诱导模型组及各加空白血清组KCs于第6小时开始出现凋亡,含高糖组KCs均较非高糖组KCs凋亡速率更快。非高糖各组与含高糖各组中促炎因子TNF-α、IL-1β含量以及炎症相关基因(TLR4、MyD88、NLRP3、Caspase-1、NF-κB/p-65、TNF-α、IL-1β、IL-18)表达水平均呈相同变化趋势,且含高糖各组上述促炎因子及炎症相关基因表达水平均较非高糖组高。以非高糖组为例,LPS诱导模型组和LPS+空白血清组的促炎因子TNF-α与IL-1β含量均显著高于对照组(P<0.05),炎症相关基因TLR4、MyD88等的mRNA表达水平也均显著高于对照组(P<0.05),而LPS诱导模型组与LPS+空白血清组间上述各指标比较差异均无统计学意义(P>0.05); 与LPS诱导模型组或LPS+空白血清组比较,LPS+化脂复肝组促炎因子TNF-α与IL-1β含量均显著降低(P<0.05),炎症相关基因TLR4、MyD88等的mRNA表达水平也均显著降低(P<0.05)。结论:化脂复肝颗粒含药血清可改善KCs活力,有效减轻KCs炎症反应,减少KCs细胞焦亡的发生,具有显著的抗炎、抗氧化损伤及保肝作用,其作用机制可能与抑制TLR4/MyD88/NF-κB p65/NLRP3信号通路的激活相关。     

Salvadora persica extract attenuates cyclophosphamide-induced hepatorenal damage by modulating oxidative stress,inflammation and apoptosis in rats

Objective: Salvadora persica(SP) is used as a food additive and is a common ingredient in folk medicine.This study investigates the antioxidant, anti-inflammatory, and beneficial effects of SP against cyclophosphamide(CYP) toxicity in rats.Methods: In a 10-day study, 32 male rats were equally allocated into 4 groups(8 rats/group) as follows:the normal control(NC group), normal rats that only received oral aqueous extract of SP(100 mg/[kg·d];SP group), animals treated with intraperitoneal CYP inj...     

小钻木脂素类化合物戈米辛R的体外抗炎机制研究

目的 通过脂多糖(LPS)诱导活化小鼠单核巨噬细胞系RAW264.7建立炎症模型,探讨小钻木脂素类化合物戈米辛R对炎症细胞增殖及炎症因子分泌的影响,评价其抗炎活性及机制。方法 采用MTT比色法检测经LPS诱导的RAW264.7细胞存活率并计算半数抑制浓度(IC₅₀);采用ELISA法检测细胞上清液TNF-α、IL-1β、IL-6等炎症因子含量;利用RT-PCR法检测细胞TNF-α、IL-1β、IL-6、IκB-α、NF-κB p65的mRNA表达量;Western blot法检测细胞IκB-α及NF-κB p65蛋白表达量。结果 与LPS模型组比较,戈米辛R可显著抑制LPS诱导的RAW264.7细胞增殖;降低细胞分泌TNF-α、IL-1β及IL-6的含量;升高LPS所致降低的IκB-α mRNA的表达量同时降低LPS引起升高的TNF-α、IL-1β、IL-6、NF-κB p65的mRNA表达量;升高IκB-α并降低NF-κB p65的蛋白表达量,显示出良好的剂量依赖性,具有统计学差异(P<0.05)。结论 戈米辛R可通过抑制TNF-α、IL-1β、IL-6、NF-κB p65的mRNA和NF-κB p65蛋白的表达,增加IκB-α mRNA和IκB-α蛋白的表达,减少致炎因子TNF-α、IL-1β、IL-6的释放,从而发挥显著抗炎活性。     

Berberine alleviates oxidized low‐density lipoprotein‐induced macrophage activation by downregulating galectin‐3 via the NF‐κB and AMPK signaling pathways

Macrophage activation plays a central role in neoatherosclerosis and in‐stent restenosis after percutaneous coronary intervention (PCI). Galectin‐3, mainly expressed on macrophages, is an important regulator of inflammation. This study aimed to investigate the effects of berberine (BBR) on oxidized low‐density lipoprotein (ox‐LDL)‐induced macrophage activation and galectin‐3 expression and their underlying mechanisms. THP‐1‐derived macrophages were pretreated with BBR prior to stimulation with ox‐LDL. Galectin‐3 expression was measured by real‐time PCR, Western blotting, and confocal microscopy. Macrophage activation was assessed by lipid accumulation, expression of inflammatory cytokines, and CD11b and CD86. Plasma galectin‐3 levels were measured in patients undergoing PCI at baseline and after BBR treatment for 3 months. BBR suppressed ox‐LDL‐induced upregulation of galectin‐3 and macrophage activation. Overexpression of galectin‐3 intervened the inhibitory effect of BBR on macrophage activation. BBR activated phospho‐AMPK and inhibited phospho‐NF‐κB p65 nuclear translocation. AMPK inhibition and NF‐κB activation abolished the inhibitory effects of BBR on galectin‐3 expression and macrophage activation. Combination of BBR and rosuvastatin exerted greater effects than BBR or rosuvastatin alone. However, BBR treatment did not further reduce plasma galectin‐3 after PCI in patients receiving standard therapy. In conclusion, BBR alleviates ox‐LDL‐induced macrophage activation by downregulating galectin‐3 via the NF‐κB and AMPK signaling pathways.     

炎症性细胞因子致肝脏缺血再灌注损伤和丹参单体的保护作用

目的：探讨肿瘤坏死因子α（TNF-α）和白细胞介素8（IL－8）在肝脏缺血再灌注损伤中的变化以及水溶性丹参单体MP－1（简称MP－1）的作用。方法：建立人肝窦内皮细胞体外低温缺氧再氧化模型和大鼠离体肝脏缺血再灌注损伤模型，以台盼蓝染色观察细胞损伤情况，用放免法分析透明质酸吸收率来反映肝AST、LDH的变化反映肝脏功能。结果：低温缺氧再氧化期间，肝窦内皮细胞的死亡率随着时间的增加而增加，TNF-α和IL－8的释放也随之增加，但肝窦内皮细胞的功能下降；肝窦内皮细胞死亡率分别与TNF-α与IL－8的释放呈正相关（r=0.949,P<0.05和r=0.892,P<0.05)。加入TNF-α抗体低温缺氧再氧化6h的内皮细胞死亡率降低；重组TNF-α处理的肝窦内皮细胞死亡率明显增加；离体大鼠肝脏的功能随低温保存和再灌注时间的增加而降低；MP－1能明显降低人肝窦内皮细胞的死亡率及TNF-α和IL－8的释放，同时可以减轻离体大鼠肝脏的缺血再灌注损伤。结论：TNF-α直接参与肝窦内皮细胞的缺血再灌注损伤，MP－1可能通过抑制TNF-α和IL－8的途径来减轻肝脏缺血再灌注损伤。     

五灵胶囊对脂多糖诱导枯否细胞内核转录因子-κB表达的作用

目的:通过体内、体外实验探讨五灵胶囊(WL)对免疫性肝脏炎症反应的作用机制。方法:制备卡介苗(BCG)+免疫佐剂+脂多糖(LPS)诱导小鼠免疫性肝损伤模型,WL灌胃给药12d,处死小鼠分离血清并制备肝组织蛋白;予大鼠WL 10、6.25g/kg灌胃4d,腹主动脉取血并制备含药血清;另取SD大鼠分离原代肝细胞和原代枯否细胞(KCs);采用Transwell小室下层培养KCs,含药血清-KCs条件培养上清对小室上层肝细胞作用。酶法测定免疫性肝损伤小鼠血清AST、ALT及条件培养上清中AST、ALT和LDH-L活性。取KCs培养成单层并同步化24h,PDTC和WL分别干预KCs 24h,再加入60μg/L的LPS诱导12h,Real-time PCR检测LPS诱导KCs表达MCP-1、IL-1、TNF-αmRNA水平,Western Blot检测免疫性肝损伤肝组织中和体外培养KCs表达TNFRⅠ、NF-κBs亚蛋白及IκKβ、IκB。结果:WL明显降低免疫性肝损伤小鼠血清ALT、AST活性并抑制肝组织内活化NF-κB信号通路;含药血清Ⅰ、Ⅱ组明显抑制活化KCs分泌促炎因子损伤肝细胞,WL抑制LPS诱导KCs表达NF-κBs信号通路蛋白,下调MCP-1、IL-1、TNF-αmRNA表达。结论:WL治疗慢性肝病炎症反应的作用机制与下调活化NF-κBs信号通路蛋白,阻滞KCs释放过量促炎因子所致肝细胞损伤相关。     

Protective effect of cinnamic acid in endotoxin‐poisoned mice

In this work, we aimed to evaluate the protective effect of cinnamic acid (CD) on lipopolysaccharide (LPS; Escherichia coli 055:B5)‐induced endotoxin‐poisoned mice and clarify the underlying mechanisms. The mice were administrated CD 5 d before 15 mg/kg LPS challenge. 12 hr later, thymus was separated for determination of thymus indexes. Lung and spleen tissues were collected for histologic examination and the wet/dry weight ratio of lung was calculated, and serum was acquired for tumor necrosis factor‐α (TNF‐α), interleukin (IL)‐18, and IL‐1β measurement. Moreover, the expression of NOD‐like receptor (NLR) family, pyrin domain‐containing 3 (NLRP3) inflammasome was determined in lung. CD increased the thymus indexes and decreased lung wet/dry weight ratio. In addition, CD improved the lung and spleen histopathological changes induced by LPS and decreased the number of neutrophils in lung tissues. CD also inhibited the pro‐inflammatory cytokines (TNF‐α, IL‐18, and IL‐1β) production in serum. Furthermore, CD suppressed the LPS‐induced NLRP3, Caspase‐1, and IL‐1β mRNA expression in lung, as well as the expression of NLRP3 and Caspase‐1 (p20) protein. CD may have protective effects in endotoxin‐poisoned mice via inhibiting the activation of NLRP3 inflammasome, and can be considered as a potential therapeutic candidate for diseases involved in endotoxin poisoning such as sepsis.     

天麻素对毛果芸香碱诱发的癫痫大鼠TLR4/NF-κB信号通路影响的研究

目的:探讨天麻素对毛果芸香碱诱发的癫痫大鼠的脑保护作用及其作用机制。方法:72只大鼠随机分为空白组、模型组、阳性药组、天麻素组、Toll样受体4 (TLR4)激活剂脂多糖(LPS)组、天麻素+LPS组,每组12只。采用腹腔注射毛果芸香碱建立癫痫大鼠模型。双抗体夹心酶联免疫法检测大鼠血清和海马组织肿瘤坏死因子-α(TNF-α)与白细胞介素-1β(IL-1β)水平,TUNEL染色法检测海马组织细胞凋亡数,Western blot分别检测海马组织活化半胱氨酸基天冬氨酸酶-3 (CleavedCaspase-3)、B淋巴细胞瘤-2 (Bcl-2)、Bcl-2相关X蛋白(Bax)、TLR4、兔抗大鼠p-核因子-κB亚基p65亲和肽(p-NF-κBp65)和磷酸化核因子κB抑制蛋白α(p-IκB-α)表达。结果:与空白组比较,模型组大鼠脑电频率显著降低(P<0.05),波幅显著升高(P<0.05);血清和海马组织TNF-α和IL-1β水平,海马组织细胞凋亡率、Cleaved-Caspase-3、Bax、TLR4和p-NF-κBp65蛋白明显升高(P<0.05);IL-10水平,Bcl-2和p-IκB-α蛋白表达显著降低(P<0.05)。与模型组比较,阳性药组、天麻素组和天麻素+LPS大鼠脑电频率显著升高(P<0.05),波幅显著降低(P<0.05),血清和海马组织TNF-α和IL-1β水平,海马组织细胞凋亡率、Cleaved-Caspase-3、Bax、TLR4和p-NF-κBp65蛋白明显降低(P<0.05),IL-10水平,Bcl-2和p-IκB-α蛋白表达显著升高(P<0.05);LPS组大鼠脑电频率、血清及海马组织IL-10水平、Bcl-2蛋白显著降低(P<0.05),血清与海马组织TNF-α与IL-1β水平、海马组织细胞凋亡率、Cleaved-Caspase-3、Bax、TLR4、p-NF-κBp65和p-IκB-α蛋白表达明显升高(P<0.05)。与LPS组比较,天麻素+LPS组大鼠脑电频率显著升高(P<0.05),波幅显著降低(P<0.05);血清及海马组织TNF-α与IL-1β水平、海马组织细胞凋亡率、Cleaved-Caspase-3、Bax、TLR4、p-NF-κBp65蛋白表达均明显降低(P<0.05),IL-10水平、Bcl-2及p-IκB-α蛋白表达明显升高(P<0.05)。结论:天麻素可通过抑制TLR4/NF-κB信号通路对毛果芸香碱诱发的癫痫大鼠发挥脑保护作用。     

何首乌水提物大鼠连续灌胃给药28 d肝毒性研究——胆汁淤积相关机制探讨

目的:考察何首乌水提物(AEPM)对大鼠肝脏中胆汁酸合成、代谢、转运相关分子影响,探讨何首乌肝毒性相关机制。 方法:SD大鼠分别灌胃AEPM 60,30 g·kg^-1,每天1次,连续28 d。28 d后解剖取肝脏,分别采用荧光定量PCR和Western blot检测肝脏MRP3,MRP2,BSEP,FXR,CYP7A1等分子的mRNA和蛋白表达水平。 结果:与正常组相比,AEPM高剂量组雄性大鼠肝脏中MRP3和BSEP的mRNA表达均显著升高(P<0.05),而AEPM高、低剂量组肝脏FXR的mRNA表达均显著降低(P<0.05);AEPM高、低剂量组雌性大鼠肝脏 MRP3,MRP2,BSEP,CYP7A1的mRNA表达均显著升高(P<0.05)。Western blot检测结果显示,AEPM高、低剂量给药组雄性和雌性大鼠肝脏中MRP3,MRP2,BSEP,FXR,CYP7A1蛋白表达水平与mRNA变化基本一致,但均未达统计学显著差异。 结论:AEPM大鼠灌胃给药28 d对肝脏胆汁酸合成、转运、排泄相关蛋白的表达具有一定的影响,在mRNA表达水平既具有胆汁淤积分子特征,同时也可见促进胆汁酸排泄的分子特征。     

二苯乙烯苷扰乱胆汁酸平衡并抑制磷脂分泌诱导肝损伤

何首乌是中国传统的中草药之一,被报道具有生发乌发、改善血脂、抗炎及抗衰老等生理活性,然而近年来何首乌诱发的肝损伤引起了广泛关注。二苯乙烯苷(2,3,5,4'-tetrahydroxystibane-2-O-β-D-glucoside,TSG)为何首乌主要成分之一,但其在何首乌肝损伤中的作用尚不清楚。该研究主要考察二苯乙烯苷诱发的肝损伤及其对胆汁酸平衡和磷脂分泌的影响。C57BL/6J小鼠腹腔注射400 mg·kg^-1TSG,检测肝损伤生化指标及磷脂含量的变化并采用LC-MS/MS技术检测胆汁酸成分的变化。结果显示,腹腔注射400 mg·kg^-1TSG显著升高血清总胆汁酸(total bile acid,TBA)、碱性磷酸酶(alkaline phosphatase,ALP)水平并显著升高游离胆汁酸含量,包括血清中的β-鼠胆酸(β-muricholic acid,β-MCA)、熊去氧胆酸(ursodeoxycholic acid,UDCA)、猪去氧胆酸(hyodeoxycholic acid,HDCA)、鹅去氧胆酸(chenodeoxycholic acid,CDCA)、去氧胆酸(deoxcholic acid,DCA)及肝脏中的β-MCA和CDCA;相关机制研究表明TSG可显著抑制法尼醇X受体(farnesoid X receptor,FXR)及下游胆酸盐外排转运蛋白(bile salt export pump,BSEP)表达导致胆汁酸蓄积,同时TSG还可抑制胆固醇25α7羟化酶(25-hydroxycholesterol-7alpha-hydroxylase,CYP7B1)干扰胆汁酸合成替代途径。另外,腹腔注射400 mg·kg^-1TSG还显著降低胆汁中磷脂含量,进一步研究发现TSG显著抑制磷脂转运蛋白多药耐药蛋白2(multidrug resistance protein 2,MDR2)的表达并破坏肝脏胆管膜上MDR2的规则分布;体外结果显示TSG在HepG2细胞上的半抑制浓度约为1500μmol·L^-1,在HepG2细胞上500μmol·L^-1TSG作用24 h可破坏胆管膜上MDR2的规则分布。综上,TSG可通过扰乱胆汁酸稳态和抑制磷脂分泌诱导肝损伤。     

The mechanism of hepatoprotective effect of sesquiterpene rich fraction from Cichorum glandulosum Boiss. et Huet on immune reaction-induced liver injury in mice

Ethnopharmacological relevance

Cichorum glandulosum Boiss. et Huet is a traditional Uygur herbal medicine that has been used as a cholagogic and diuretic agent to improve liver function. However, the mechanism is not known for the liver-protective function. We investigated the antioxidant effects of plant extraction (CGE60) in vitro and in vivo, and find the mechanism of liver protection in Bacille Calmette-Guerin vaccine (BCG)+Lipopolysaccharides (LPS) induced liver injury in mice.

Materials and methods

CGE60 was made, and the antioxidant activity was investigated by comparing the ability of scavenging 1,1-diphenyl-2-picrylhydrazyl (DPPH), and 2,2-azinobis(3-ehtylbenzothiazolin-6-sulfnicAcid) diammonium salt (ABTS) free radicals in vitro. Then, CGE60 was administrated in mice of liver damage model which was induced in mice using the BCG+LPS protocol. The CGE 60 extract was tested at three dosages: 50 mg/kg, 100 mg/kg, and 200 mg/kg. Product of lipid peroxidation (MDA), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-PX,), nitric oxide (NO), nitric oxide synthetase (NOS), hydroxyproline and alkaline phosphatase (ALP) contents were evaluated in liver to determine the CGE60 activity in the hepatic injury model. Tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and transforming growth factor-β (TGF-β) proteins were determined in the liver tissues using ELSIA. The signaling activities were evaluated in Western blot.

Results

CGE60 exhibited strong antioxidant ability in vitro. With oral administration, CGE60 significantly increased the activity of CAT, SOD, GSH-PX, and decreased the level of NO, NO synthase, hydroxyproline, ALP and lipid peroxidation liver of in the BDG+ LPS model. CGE60 attenuated hepatic inflammation via down- regulation of TNF-α, IL-6 and TGF-β. CGE60 blocked protein expression of cytochrome P450 2E1 (CYP2E1), nuclear factor kappa-B (NF-κB), phosphorylation of extracellular signal-regulated kinase (p-ERK1/2), and cyclooxygenase-2 (COX-2),but activated the expression of p-P38 MAPK.

Conclusion

This study suggests that CGE60 possesses antioxidant activity and this activity associates with hepatoprotective effect in the mice of BCG +LPS model, and the mechanisms underlying these effects may involve antioxidant actions and anti-inflammation activities.     

高效快速降脂动物模型研究

探讨斑马鱼作为降脂药物筛选模型的可行性以及盐酸小檗碱(BBR)对高脂斑马鱼的影响。用含4%胆固醇饲料喂养3月龄斑马鱼0,2,4,8,14,20,25,30 d,检测血清总胆固醇含量的变化;测定对照组、高胆固醇组、0.01%辛伐他汀组、0.1%BBR组和0.2%BBR组斑马鱼血清总胆固醇(TC)、总甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-c)和高密度脂蛋白胆固醇(HDL-c)的水平;RT-PCR检测肝脏HMGCR,LDLR和CYP7A1a mRNA的表达水平;油红O染色检测肝脏中脂肪含量的变化。结果显示4%胆固醇喂养斑马鱼后血清TC水平升高且有时间依赖性,20 d基本达到稳定水平;BBR组斑马鱼血清TC,TG和LDL-c水平显著下降,肝脏HMGCR mRNA的表达下降,LDLR和CYP7A1a mRNA的表达升高,脂肪含量减少。4%胆固醇饲料喂养斑马鱼20 d即可建立高脂斑马鱼模型,研究结果为进一步开展降脂药物的筛选奠定基础。     

多反应监测方法评估丹参和人参对大鼠肝药酶的调控作用

目的建立新型特异性强的高通量细胞色素P450(CYP)丰度测定方法,以人参和丹参为工具药考察药物对肝药酶的调控作用。方法采用基于质谱多反应监测(MRM)模式方法,以串联配体法(QconCAT)表达合成的重标肽段作为内标对丹参与人参处理后,对9种大鼠肝脏CYP(CYP1A1、CYP1A2、CYP2B1、CYP2B2、CYP2C6、CYP2C11、CYP3A1、CYP3A2、CYP17A1)蛋白特异肽段进行定量,探讨丹参和人参对肝脏CYP酶的表达上调或者下调作用。结果 QconCAT重标合成内标肽段与肝脏样品肽段质谱表征一致,与对照组相比,丹参与人参作用后,对大鼠肝内CYP1A1、CYP2B2的表达均具有下调作用,对CYP1A2、CYP2B1的表达均具有上调作用。丹参同时下调CYP3A2、CYP2C11和CYP17A1的表达,而人参则上调这3种蛋白的表达。建立了MRM方法对大鼠肝脏CYP蛋白定量方法,定量方法相对标准偏差在5.9%以内,相对误差在6.8%以内,定量标准曲线的线性系数大于0.9。结论对9种大鼠肝脏CYP酶进行定量研究,其中CYP2B1、CYP2B2和CYP17A1是首次进行蛋白质定量研究。丹参与人参对CYP亚型产生的调控作用存在差异性,为药物配伍实践中提供临床参考价值,避免不良药物相互作用反应。     

Isoforskolin and forskolin attenuate lipopolysaccharide‐induced inflammation through TLR4/MyD88/NF‐κB cascades in human mononuclear leukocytes

The principal active component of isoforskolin (ISOF) is from the plant Coleus forskohlii, native to China, which has attracted much attention for its biological effects. We hypothesize that ISOF and forskolin (FSK) pretreatment attenuates inflammation induced by lipopolysaccharide (LPS) related to toll‐like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), and nuclear factor kappa B (NF‐κB) signaling. Mononuclear leukocytes (MLs) from healthy donors' blood samples were separated by using density gradient centrifugation. Protein levels of TLR4, MyD88, and NF‐κB were detected using western blot and inflammatory cytokines interleukin (IL) 1β, IL‐2, IL‐6, IL‐21, IL‐23, tumor necrosis factor (TNF) α, and TNF‐β were tested by enzyme‐linked immunosorbent assay and Quantibody array in MLs. Our results showed that LPS augmented the protein levels of TLR4, MyD88, and NF‐κB in MLs and the production of IL‐1β, IL‐2, IL‐6, IL‐21, IL‐23, TNF‐α, and TNF‐β in supernatants of MLs. Despite treatment with ISOF and FSK prior to LPS, the protein levels of TLR4, MyD88, NF‐κB, IL‐1β, IL‐2, IL‐6, IL‐21, IL‐23, TNF‐α, and TNF‐β in MLs were apparently decreased. roflumilast (RF) and dexamethasone (DM) had a similar effect on MLs with ISOF and FSK. Our results, for the first time, have shown that ISOF and FSK attenuate inflammation in MLs induced by LPS through down‐regulating protein levels of IL‐1β and TNF‐α, in which TLR4/MyD88/NF‐κB signal pathway could be involved.     

脑心通调控NLRP3/Caspase-1通路抑制脂多糖诱导的BV2细胞焦亡

目的:探索脑心通醇提物对脂多糖(LPS)诱导的小胶质细胞BV2焦亡的影响,并通过NOD样受体热蛋白结构域3(NLRP3)/半胱氨酸蛋白酶-1(Caspase-1)通路阐释其可能的作用机制。方法:LPS(1 mg·L^-1)体外诱导BV2细胞的同时,加入不同质量浓度脑心通醇提物(2,10,50 mg·L^-1)干预后,分别采用MTS法检测细胞活性,实时荧光定量PCR(Real-time PCR)检测白细胞介素-1β(IL-1β),肿瘤坏死因子-α(TNF-α)和NLRP3的mRNA水平;免疫荧光法检测NLRP3蛋白表达的情况;蛋白免疫印迹法(Western blot)检测NLRP3/Caspase-1通路关键蛋白表达。结果:与空白组比较,LPS(1 mg·L^-1)能明显降低BV2细胞活性,显著升高IL-1β,TNF-α及NLRP3 mRNA表达水平(P<0.01),且可提升NLRP3蛋白表达水平以及Caspase-1剪切体和前体蛋白的比值(Caspase-1 p20/Caspase-1)。与LPS组比较,脑心通醇提物(2,10,50 mg·L^-1)干预后,逆转了LPS导致的细胞活性降低(P<0.01),50 mg·L^-1脑心通醇提物可显著降低IL-1β,TNF-α及NLRP3 mRNA表达水平(P<0.05,P<0.01),显著降低LPS诱导的NLRP3的高表达以及Caspase-1 p20/Caspase-1(P<0.01)。结论:脑心通能明显抑制LPS诱导的小胶质细胞焦亡,其作用机制与NLRP3/Caspase-1通路密切相关。     

酒肝宁煎剂对酒精性肝损伤大鼠CYP2E1及白介素6、肿瘤坏死因子-α含量的影响

目的 观察中药酒肝宁煎剂对酒精性肝损伤大鼠CYP2E1及细胞因子白介素-6(IL-6),肿瘤坏死因子-α(TNF-α)含量的影响.方法 清洁级Wistar雄性大鼠72只,根据大鼠体重,采用分层抽样法将大鼠分为6组,空白对照组、酒肝宁毒性试验组、酒精性肝病模型组、益肝灵治疗组、酒肝宁低剂量组、酒肝宁高剂量组.空白对照组灌...