葛根素减轻LPS诱导的小鼠肾脏炎性反应

目的 探讨葛根素(PUE)是否减轻脂多糖(LPS)诱导的小鼠炎性反应。方法 选择30名健康体检者为对照(control)组,45例严重感染(SI)非肾损伤患者作为N-SI组,30例严重感染致肾损伤患者作SI组,收集患者外周血样本。将50只小鼠随机分为对照(Ctrl)组,LPS组,LPS+PUE(10、25、50 mg/kg·d, ip)组,PUE在给予LPS后1 h应用,24 h后处死小鼠,采集血液。ELISA检测血清Scr、BUN、IL-6和IL-33水平。结果 与对照组和N-SI组相比,SI组患者血清中BUN、Scr、IL-6和IL-33水平显著升高(P<0.05)。小鼠LPS组与对照组相比,血清中肾功能指标BUN和Scr均显著增高(P<0.05);与LPS组相比,PUE可显著降低BUN、Scr、MDA、IL-33和IL-6的表达水平,并使SOD的活性升高(P<0.05)。结论 PUE可减轻LPS诱导的小鼠炎性反应。     

Lactobacillus casei Zhang 对扑热息痛诱导的小鼠急性肝损伤的保护作用

目的:研究益生菌Lactobacillus casei Zhang(Lcz)对扑热息痛(APAP)所致小鼠急性肝损伤的保护作用及其机制。方法:C57BL/6 小鼠被随机分为空白组(Ctrl)、APAP 诱导急性肝损伤模型组(Acetaminophen ,APAP)、阳性药物组(N-Acetylcysteine,NAC)、Lcz 预防组(Lcz/ APAP)和Lcz 对照组(Lcz)。Lcz(1伊109 CFU/ ml)连续灌胃30 d 后,NAC 组在APAP 处理前1 h 腹腔注NAC(150 mg/ kg)。APAP、NAC 以及Lcz/ APAP 组均腹腔注射APAP(300 mg/ kg)。APAP 作用18 h 后,采集血液和收集肝脏组织,检测血清中谷丙转氨酶(ALT)和谷草转氨酶(AST)的水平。通过Western blot 检测肝脏组织中血红素氧化酶(HO-1)、超氧化物歧化酶2(SOD2)、Bcl-2 以及TLR4 的表达水平。结果:Lcz 能抑制APAP 诱导的急性肝损伤血清中ALT 和AST 水平。Lcz 提高了HO-1、SOD2 和Bcl-2 的蛋白表达水平,而降低了APAP 诱导的TLR4 的表达。结论:益生菌Lcz对APAP 诱导的小鼠急性肝损伤有保护作用,其保肝作用机制可能与其抗氧化和抗炎活性有关。     

环磷腺苷葡胺和氨茶碱对小鼠急性缺氧性脑损伤的保护作用

目的观察环磷腺苷葡氨（MCA）和氨茶碱联用对小鼠急性缺氧性脑损伤（HBI）的干预作用，初步探讨其可能的神经保护机制。方法90只小鼠分为正常对照组、HBI组、氨茶碱组、MCA组、两药合用组。缺氧后72h测各组脑组织含水量、超氧化物歧化酶活性（SOD）及氧化代谢产物丙二醛（MDA）含量；HE染色观察各组海马区结构及神经细胞的调亡情况。结果与HBI模型组相比，药物干预组脑组织含水量、MDA下降，SOD升高，病理变化减轻，细胞调亡减少，病理变化尤以两药合用组改善最显著。结论两药联用可有效抑制缺氧后脑组织内氧自由基生成，减轻脑水肿及神经细胞的调亡。     

红藤对LPS诱导流产小鼠的保胎作用及子宫巨噬细胞的影响

目的:探讨红藤对细菌脂多糖(LPS)诱导流产小鼠的保护作用及其免疫药理机制。方法:昆明种小鼠55只,随机分为对照组(A组,10只)和实验组。实验组又分为LPS处理组(B组)、红藤处理组(C组)、红藤及LPS双处理组(D组),每组各15只。观察妊娠结果,并用酶组织化学、免疫组化、ELISA方法检测子宫非特异性酯酶阳性(α-NAE+)、CD14+、CD204+巨噬细胞和TNF-α的变化。结果:小鼠流产率在B组为100%(P<0.01),胚胎吸收率为100%(P<0.01);而D组流产率降低为13.33%,胚胎吸收率降至12.56%;孕鼠子宫α-NAE+、CD14+巨噬细胞数量,B组子宫各部位与A组相比均显著增多(P<0.01);D组环肌外侧与A组相比显著增多(P<0.01),但环肌内侧、功能层无明显变化,C、D组CD204+巨噬细胞数量与A组、B组相比均显著增多(P<0.01);子宫TNF-α含量B组与A组相比显著增加(P<0.01),而D组接近正常水平。结论:红藤可以对抗LPS所致的小鼠流产,通过影响孕鼠子宫巨噬细胞的数量、分布和亚群,抑制TNF-α的分泌,是其保护胚胎正常发育的机理之一。     

模拟肽对LPS诱导CD14的表达

本文观察PMB及其模拟肽处理前后FITC-LPS与J774A1的结合能力及CD14表达,并检测培养上清中细胞因子TNF-α、IL-6的含量变化。结果发现①FITC-LPS分别与PMB、peptide 1孵育后,细胞膜平均通道荧光显著减少,LPS与J774A1的结合能力显著降低;②100 ng/ml LPS刺激J774A1 3 h后CD14阳性率明显增加;LPS分别与PMB和peptide 1预孵育后可显著降低LPS刺激J774A1细胞膜CD14表达;③LPS刺激J774A1分泌TNF-α和IL-6显著增加,LPS分别与PMB和petide 1预孵育后能显著减少细胞因子TNF-α和IL-6分泌。结果证实多粘菌素B及其模拟肽(peptide 1)可能通过下调J774A1 CD14表达,降低细胞因子水平来减少LPS诱导的炎症反应。     

葛根素对小鼠脑微循环血流量的影响

为了观察静脉注射葛根素对小鼠脑微循环的影响，应用ＪＩ ２００型激光微循环动态分析仪直接记录脑微循环血流量的变化。结果表明，正常小鼠静脉注射葛根素（０．２ｍｇ／ｋｇ）３ｍｉｎ后，脑微循环血流量显著增加（Ｐ＜０．０５），且持续３０ｍｉｎ；提高葛根素剂量（２ｍｇ／ｋｇ），其效应加强且持续６０ｍｉｎ。高分子右旋糖酐所致脑微循环障碍小鼠静脉注射葛根素后，脑微循环血流量有改善，但作用较弱。结果提示，葛根素可增加正常小鼠脑微循环血流量，改善脑微循环     

葛根素抑制乙醇诱导的小鼠骨髓细胞成脂基因表达

乙醇能够诱导骨髓基质细胞（marrow stromal cells,MSCs）大量分化为脂肪细胞,与乙醇性股骨头坏死密切有关。本研究旨在观察葛根素掏乙醇导致MSCs成脂变化的基因表达。     

LPS诱导的内毒素血症小鼠及重症联合免疫缺陷小鼠模型炎症反应的比较

目的:比较脂多糖(LPS)诱导的内毒素血症BALB/c小鼠及重症联合免疫缺陷(SCID)小鼠炎症反应的差异。方法:建立LPS诱导的BALB/c小鼠和SCID小鼠内毒素血症模型,观察小鼠的存活率。分别于诱导前、诱导后3h、6h、12h,取小鼠的血清及诱导后12h小鼠的肝脏、肺脏,用全自动生化分析仪检测两种小鼠血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、尿素氮(BUN)水平;HE染色评价肝脏、肺脏的炎症病理改变;用流式细胞术微球阵列法检测两种小鼠血清TNF-α、IFN-γ、IL-6及MCP-1的水平。结果:(1)LPS诱导内毒素血症后,SCID小鼠于12～24h均死亡(8/8),BALB/c小鼠仅1只死亡(1/8)。(2)LPS诱导后12h,BALB/c小鼠及SCID小鼠血清ALT、AST、BUN的水平均明显升高(P<0.05),SCID小鼠前两项均高于BALB/c小鼠(P<0.05),但BUN两种小鼠无显著差异。(3)肺脏,肝脏炎症盲法的病理评分,SCID小鼠均高于BALB/c小鼠(P<0.05)。(4)SCID小鼠和BALB/c小鼠LPS诱导后3h、6h、12h,血清TNF-α,IFN-γ的水平,诱导后12h,IL-6,MCP-1的水平均显著升高(P<0.05),SCID小鼠明显高于BALB/c小鼠(P<0.05)。结论:LPS刺激SCID小鼠后,可分泌过量的炎性细胞因子,导致更严重的内毒素血症和脏器损伤,是造成小鼠死亡的重要原因。结果提示,缺乏适应性免疫应答细胞调控的情况下,异常增强的固有免疫应答,可能是危及机体生命的重症全身炎症反应综合征发生的重要原因。     

姜黄素对癫痫小鼠海马脑损伤的保护作用

目的研究姜黄素对匹罗卡品诱导的癫痫小鼠海马新生神经元和细胞凋亡的影响。方法姜黄素预处理后,小鼠腹腔注射匹罗卡品建立小鼠癫痫模型,应用新生神经元标记物双皮层蛋白(doublecortin,DCX)免疫组织化学染色及TUNEL染色对造模后72h的模型小鼠海马进行检测。结果 DCX免疫组织化学染色结果表明,与对照组相比,模型组及姜黄素处理模型组小鼠海马齿状回DCX阳性细胞明显减少;与模型组相比,姜黄素处理模型组小鼠海马齿状回DCX阳性细胞明显增多。TUNEL染色结果表明,与对照组相比,模型组及姜黄素处理模型组小鼠海马齿状回TUNEL阳性细胞明显增多;与模型组相比,姜黄素处理模型组小鼠海马齿状回TUNEL阳性细胞明显减少。结论姜黄素可能对匹罗卡品诱导的癫痫小鼠海马神经元有保护作用。     

雷公藤甲素对LPS诱导的海马炎症过程中COX-2和iNOS表达的影响

选用健康成年雄性SD大鼠,用不同剂量(10μg·kg-1·d-1,25μg·kg-1·d-1,50μg·kg-1·d-1)的雷公藤甲素预处理5d后,立体定位海马注射脂多糖(LPS),采用免疫组织化学染色方法观察海马CA3区环氧化酶2(COX-2)和诱导型一氧化氮合酶(iNOS)的表达变化。结果发现:与注射生理盐水对照组比较,注射LPS可引起海马CA3区的COX-2和iNOS显著表达,而雷公藤甲素(50μg·kg-1·d-1)可明显下调LPS诱导的COX-2和iNOS的表达,其抑制程度与药物剂量呈正相关。本实验结果提示雷公藤甲素对中枢神经系统中LPS诱导的COX-2和iNOS的表达有明显的抑制作用。     

高压氧对小鼠脑缺血再灌注损伤的保护作用

目的探讨高压氧（HBO）对脑缺血再灌注小鼠神经元的保护作用。方法选取昆明小鼠27只，采用无创微动脉夹阻断双侧颈总动脉血流的方法建立脑缺血再灌注模型。采用HE染色、透射电镜观察脑缺血再灌注小鼠神经元的损伤及HBO干预后的改变，结果光镜显示3组小鼠额叶皮层及海马有不同程度的神经细胞变性、坏死；实验组与对照组异常细胞数比较羞异有统计学意义（P〈0．05）：同组小鼠的皮质和海马比较差异无统计学意义（P〉0．05）。电镜观察显示再灌注后额叶皮质及海马有不同程度的神经元变性、坏死，实验组损害明显减轻，与对照组比较差异有统计学意义（P〈0．05）；同组小鼠的皮质和海马损伤程度比较差异无统计学意义（P〉0．05）。结论HBO治疗可对小鼠脑缺血再灌注损伤具有保护作用．可促进损伤的修复。     

Chikusetsusaponin V attenuates lipopolysaccharide-induced liver injury in mice

Chikusetsusaponin V (CsV), a saponin from Panax japonicus, has been reported to inhibit inflammatory responses in lipopolysaccharide (LPS)-induced macrophage cells. However, whether CsV could alleviate LPS-induced liver injury in vivo and the potential mechanisms involved remain unclear. In the present study, we investigated the anti-inflammatory effects of CsV on LPS-induced acute liver injury in mice and further explored the potential mechanisms involved. Our results showed that CsV significantly attenuated elevation of alanine transaminase (ALT) and aspartate aminotransferase (AST) levels and improved liver histopathological changes in LPS-induced mice. In addition, CsV decreased serum tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) levels and inhibited mRNA expressions of inducible nitric oxide synthase (iNOS), TNF-α and IL-1β in LPS challenged mice. Furthermore, CsV inhibited nuclear factor kappa B (NF-κB) activation by downregulating phosphorylated NF-κB, IκB-α, ERK, c-Jun N-terminal kinase (JNK) and p38 levels in the liver tissue, which ultimately decreased nucleus NF-κB protein level. In conclusion, our data suggested that CsV could be a promising drug for preventing LPS challenged liver injury since it attenuated LPS-induced inflammatory responses, partly via inhibiting NF-κB and MAPK signaling pathways.     

胍丁胺对内毒素诱导的大鼠急性肺损伤的治疗效果

目的 :研究胍丁胺对内毒素(LPS)诱导的急性肺损伤治疗效果及其机制。方法 :采用LPS诱导大鼠急性肺损伤模型并予以胍丁胺治疗,分为对照组,单纯模型组,低、高剂量胍丁胺治疗组。绘制Kaplan-Meier生存曲线观察胍丁胺治疗后大鼠生存率差异,并检测肺组织湿干比及组织炎症情况;检测肺组织中抗氧化酶(SOD)活性、丙二醛(MDA)含量;收集肺泡灌洗液并提取肺组织总蛋白,检测肺组织及肺泡灌洗液中炎症因子白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)含量。结果 :胍丁胺可提高LPS诱导的大鼠急性肺损伤模型的生存率、显著降低肺组织的湿重/干重比。H-E染色结果显示胍丁胺可改善肺组织炎症状况。与单纯模型组相比,低、高剂量胍丁胺治疗组的肺组织MDA含量下降、SOD活性上调,差异具有统计学意义。ELISA结果表明,低、高剂量胍丁胺治疗组的肺组织及肺泡灌洗液中IL-6、TNF-α分泌降低。结论 :胍丁胺可抑制肺部过度氧化应激及炎症反应,发挥对急性肺损伤的保护作用。     

Prophylaxis with α-lipoic acid against lipopolysaccharide-induced brain injury in rats

Introduction  Lipopolysaccharide (LPS) stimulates the synthesis and release of reactive oxygen species that play an important role in the pathogenesis of tissue injuries. In this study the effect of early administration of the antioxidant α-lipoic acid (α-LA) on brain lipid peroxidation, brain hydrogen peroxide (H₂O₂) concentration, and brain total sulfhydryl group (-SH group) content was evaluated in rats with endotoxic shock induced by administration of LPS (Escherichia coli 026:B6, 30 mg/kg i.v.) Materials and Methods  Rats were treated intravenously with normal saline or α-LA (60 mg/kg) 30 min after LPS injection. After 5 h of observation, the animals were killed and their brains were isolated for the measurements. Results  Injection of LPS alone resulted in the development of shock and oxidative stress, the latter indicated by a significant increase in brain concentrations of thiobarbituric acid-reacting substances (TBARS) and H₂O₂ and a decrease in total brain -SH group content. Administration of α-LA after the LPS challenge resulted in an increase in total -SH group content and a decrease in TBARS and H₂O₂ concentration in the brain tissue compared with the LPS group. Conclusion  The results indicate that α-LA treatment effectively protected the brain tissue against endotoxin-induced oxidative stress. Administration of LA could be a useful adjunct to clinical application in the management of septic shock.     

Protective effect of gossypol on lipopolysaccharide-induced acute lung injury in mice

Objective

Gossypol has been reported to have anti-inflammatory properties. The purpose of this study was to evaluate the effect of gossypol on acute lung injury (ALI) induced by lipopolysaccharide (LPS) in mice.

Methods

Male BALB/c mice were pretreated with gossypol 1 h before intranasal instillation of LPS. Then, 7 h after LPS administration, the myeloperoxidase in histology of lungs, lung wet/dry ratio and inflammatory cells in the bronchoalveolar lavage fluid (BALF) were determined. The levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) in the BALF were measured by ELISA. The extent of phosphorylation of IκB-α, p65 NF-κB, p46–p54 JNK, p42–p44 ERK, and p38 were detected by western blot.

Results

Gossypol markedly attenuated the LPS-induced histological alterations in the lung and inhibited the production of TNF-α, IL-1β and IL-6. Additionally, gossypol reduced the inflammatory cells in BALF, decreased the wet/dry ratio of lungs and inhibited the phosphorylation of IκB-α, p65 NF-κB, p46–p54 JNK, p42–p44 ERK, and p38 caused by LPS.

Conclusion

The data suggest that anti-inflammatory effects of gossypol against the LPS-induced ALI may be due to its ability of inhibition of the NF-κB and MAPKs signaling pathways. Gossypol may be a promising potential therapeutic reagent for ALI treatment.     

藤黄酸减轻脂多糖诱导的小鼠急性肺损伤

目的探讨藤黄酸(GA)对脂多糖(LPS)所致小鼠急性肺损伤的保护作用及其机制。方法采用尾静脉注射LPS(4 mg/kg)建立小鼠急性肺损伤模型。实验将小鼠随机分为对照组(control组)、模型组(model组)、藤黄酸组(GA组)和藤黄酸预处理组(GA+LPS组),6 h后测定肺湿/干重比值(W/D);检测髓过氧化物酶(MPO)活性;检测肺泡灌洗液(BALF)中蛋白含量和白细胞计数;ELISA检测肺匀浆中白介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)含量。结果模型组小鼠肺W/D、MPO活性、BALF中蛋白含量和白细胞数量均增加,肺组织IL-1β和TNF-α水平升高(均P0.01);藤黄酸预处理可减轻LPS引起的以上指标变化(均P0.05)。结论 GA可减轻LPS诱导的急性肺损伤,其机制可能与降低肺组织IL-1β和TNF-α的含量、抑制中性粒细胞在肺部的聚集和减轻肺部水肿相关。     

Breastfeeding Might Have Protective Effects on Atopy in Children With the CD14C-159T CT/CC Genotype

Breastfeeding is widely recommended to reduce risk of sensitization, eczema and asthma. However, the role of breastfeeding in prevention of allergic diseases is uncertain. We aimed to investigate whether the relationship between breastfeeding and sensitization to aeroallergens is modified by cluster of differentiation 14 (CD14) genotype. This study included 1,828 school children aged 9-12. We administered a detailed questionnaire and genotyped the CD14C-159T polymorphism. Skin prick tests for 12 aeroallergens were performed. School children who had been breastfed were less likely sensitized to aeroallergens (adjusted odds ratio [aOR] 0.712, 95% confidence interval [CI]: 0.555-0.914). There was no significant association between CD14C-159T genotype and atopy. Breastfeeding was associated with a decreased risk of atopic sensitization in children with CT/CC genotype (aOR 0.667, 95% CI: 0.463-0.960). Our data might identify the gene-environment interaction between the CD14C-159T polymorphism and breastfeeding in relation to aeroallergen sensitization.     

Expression of scavenger receptor class A and CD14 in lipopolysaccharide-induced lung injury

CD14 and macrophage scavenger receptor class A type I and II (MSR-A) are receptors for lipopolysaccharide (LPS). In this study, the expressions of both receptors in the lung after administration of LPS in aerosol to mice with a nebulizer were observed. Bronchiolar epithelial cells and alveolar macrophages immediately incorporated LPS and expressed CD14. CD14-positive neutrophils then appeared in the alveolar space followed by the appearance of MSR-A-expressing cells in the vascular lumen, pulmonary interstitium, and alveolar space. Numbers of apoptotic cells increased after 1 day, and MSR-A-expressing macrophages actively incorporated apoptotic bodies. Daily administration of macrophage colony stimulating factor (M-CSF) to the mice resulted in increased levels of MSR-A expression and reduced levels of CD14 as well as several cytokine expressions, leading to shortening of the inflammatory process. The numbers of apoptotic cells were reduced in M-CSF injected mice. These findings imply that CD14 acts as an immediate expressing receptor for LPS and MSR-A exerts a protective function by scavenging LPS and apoptotic cells in LPS-induced lung injury.     

中药四毒清抑制LPS性肾脏损伤的机制研究

目的：研究中药复方四毒清防治内毒素性肾功能衰竭的作用机制。 方法： 将小鼠随机分成对照组、LPS组、四毒清防治组和四毒清组，用水（0.2 mL/10 g BW）或四毒清（1 000 g/L, 0.2 mL/10 g BW）灌胃3 d，每天2次, 第3 d灌胃后2 h，腹腔注射LPS(30 mg/kg，0.2 mL/10 g BW)或生理盐水（0.2 mL/10 g BW），腹腔注射后2 h，再用水或四毒清（0.2 mL/10 g）灌胃1次。测定各组小鼠血清肌酐（Cr）和尿素氮（BUN）的含量，观察肾脏超微结构，肾组织丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性的变化， 并用半定量RT-PCR方法测定肾组织细胞间黏附分子-1(ICAM-1) mRNA的表达。 结果： LPS引起小鼠血清Cr和BUN含量明显升高，肾脏近曲小管出现明显病理改变。四毒清有效降低LPS攻击小鼠血清Cr和BUN的含量，明显减轻近曲小管的损伤。LPS组小鼠肾组织MDA含量和ICAM-1 mRNA的表达显著高于对照组，而四毒清防治组肾组织MDA含量和ICAM-1 mRNA的表达明显低于LPS组，四毒清处理能显著升高肾组织SOD的活性。 结论： 中药四毒清防治内毒素性肾功能衰竭的作用机制与其升高肾组织SOD的活性、减轻肾组织氧化损伤并抑制肾脏ICAM-1 mRNA的表达有关。