嗜酸乳酸杆菌对自主活动受限型应激小鼠肠道黏膜免疫状态的影响

目的：探讨嗜酸乳酸杆菌（Lactobacillus acidophilus,LA）对自主活动受限型应激小鼠肠道黏膜免疫状态的影响。方法：以健康雌性BALB/c小鼠为受试动物,分为常规饲养组（NC组）、常规饲养条件灌胃LA组（NC＋LA组）、应激组（S组）、应激条件灌胃LA组（S＋LA组）,小鼠的LA灌胃剂量为108 CFU/d;应激组小鼠每天被限制自主活动3 h;15 d后取材,采用流式细胞术、酶联免疫吸附（enzyme-linked immunosorbent assay,ELISA）等方法,对肠系膜淋巴结（mesenteric lymph nodes,MLN）中CD4＋T细胞、CD8＋T细胞和CD11c＋树突细胞（CD11c＋ dendritic cell,CD11c＋ DC）比例,结肠组织白细胞介素-10（interleukin-10,IL-10）和白细胞介素-17（interleukin-17,IL-17）水平,以及小肠总分泌型免疫球蛋白A（secreted immunoglobulin A,sIgA）水平进行检测。结果：常规饲养条件下,LA能极显著提高小鼠MLN中CD4＋T细胞比例（P＜0.01）,显著提高MLN中CD11c＋ DC比例、小肠总sIgA水平和结肠组织IL-10水平（P＜0.05）,显著降低结肠组织IL-17水平（P＜0.05）,而对MLN中CD8＋T细胞比例无显著影响（P＞0.05）;应激条件能极显著降低小鼠MLN中CD4＋T细胞比例和CD11c＋ DC比例（P＜0.01）,显著降低CD8＋T细胞比例（P＜0.05）,极显著降低小鼠结肠组织IL-10水平和小肠总sIgA水平（P＜0.01）,极显著提高结肠组织IL-17水平（P＜0.01）;应激条件下,LA能显著提高小鼠MLN中CD4＋T细胞比例和CD11c＋ DC比例（P＜0.05）,显著提高小鼠结肠组织IL-10水平和小肠总sIgA水平（P＜0.05）,并显著降低结肠组织IL-17水平（P＜0.05）,而对小鼠MLN中CD8＋T细胞比例无显著影响（P＞0.05）。结论：给予自主活动受限型应激小鼠LA干预,可提高小鼠MLN中CD4＋T细胞和CD11c＋ DC比例,上调结肠IL-10和小肠总sIgA分泌水平,下调结肠IL-17分泌水平,调节应激小鼠的肠道黏膜免疫状态。     

槐花多糖对免疫抑制小鼠免疫功能的影响研究

研究槐花多糖对免疫低下小鼠的免疫功能的调节作用。采用腹腔注射环磷酰胺制造免疫抑制小鼠模型,考察槐花多糖对免疫抑制小鼠脏器指数、巨噬细胞吞噬功能、淋巴细胞增殖活性、血清IL-2、IL-4、IL-6、TNF-α、TNF-γ水平、溶血素水平和溶血空斑形成数量的影响。结果表明槐花多糖能明显提高免疫抑制小鼠的胸腺指数、脾脏指数、巨噬细胞吞噬能力、脾淋巴细胞的增殖能力、溶血素含量和溶血空斑形成数量,促进细胞因子IL-2、IL-4、IL-6、TNF-α、TNF-γ分泌。槐花多糖具有免疫增强活性,具有良好的开发应用潜力。     

液体发酵的桑黄多糖对小鼠免疫功能的影响

研究液体发酵分离纯化得到的桑黄多糖对小鼠免疫功能的影响。研究桑黄多糖对小鼠的免疫器官、碳廓清率以及IL-1、IL-2、IL-4、IL-6、IFN-γ、TNF-α细胞因子的影响,发现桑黄多糖能显著提高小鼠胸腺重量和脾脏重量,增强小鼠碳廓清率,增加IL-2和IFN一γ的含量,降低IL-1、IL-6、TNF-α的含量。证实桑黄多糖能显著增强小鼠的机体免疫力,有一定的抗炎作用。     

含酸奶饲料对衰老小鼠某些免疫功能指标的影响

用含Lactobacillus bulgaricus和Streptococcus Thermophilus活菌酸奶30％的饲料饲喂衰老小鼠3d，测定其胸腺重量、脾淋巴细胞增殖能力、IL-2（Interleukin，IL）产生量、脾T淋巴细胞中CD4^＋／CD8^＋细胞比例等指标，发现饲喂酸奶显著增加了胸腺重和IL-2的产生量,提高了脾T淋巴细胞的增殖能力，但未达到年轻小鼠的水平。此外还观察到酸奶处理提高了T淋巴细胞中CD4^＋／CD8^＋细胞的比例。这些结果表明，酸奶对于老年小鼠的免疫功能具有一定恢复作用。     

仿刺参雌性生殖腺皂苷的免疫增强活性研究

目的:以仿刺参雌性生殖腺皂苷（gonad saponins from female of Apostichopus japonicus,AGS）为研究对象,探究其免疫增强活性。方法:通过对巨噬细胞吞噬能力和细胞因子的测定,研究AGS的体外免疫调节活性;通过对免疫低下小鼠胸腺、脾脏指数,碳廓清指数与吞噬指数,腹腔巨噬细胞增殖活性,脾细胞增殖活性,自然杀伤细胞（natural killer cell,NK cell）活力,T淋巴细胞亚群水平及小鼠血清细胞因子的测定,分析AGS在小鼠体内的免疫调节活性。结果:体外实验表明,AGS能提高小鼠体外巨噬细胞吞噬能力,促进TNF-α、IL-6、IFN-γ的分泌,且各组与正常组相比差异显著（P<0.05）;体内实验表明,与模型组相比,中剂量组和高剂量组AGS均能显著提高小鼠胸腺、脾脏指数、碳廓清指数和吞噬指数（P<0.05）,显著增强腹腔巨噬细胞和脾细胞增殖活性（P<0.05）,显著增强NK细胞的活性（P<0.05）,显著提高CD4+/CD8+水平（P<0.05）,显著促进TNF-α、IL-6、IFN-γ的分泌（P<0.05）。结论:AGS在体外和体内都表现出增强免疫的活性。     

枸杞粉及其多糖对环磷酰胺致免疫低下小鼠免疫及肠道菌群的调节作用

枸杞是我国传统的药食两用资源,其鲜果喷雾干燥粉作为食品或功能食品原料被广泛应用,但其功效作用研究较少。本研究以环磷酰胺（cyclophosphamide,CTX）诱导免疫低下的小鼠为模型,探究枸杞粉（Lycium barbarum L. powder,LB）、枸杞多糖（Lycium barbarum L. polysaccharides,LBP）及复配物（LB＋LBP）对小鼠免疫力及肠道菌群稳态的影响。结果表明,LB、LBP及LB＋LBP均可不同程度地恢复免疫抑制小鼠的体质量和采食量,改善小鼠的脾脏、胸腺和结肠组织损伤,有效促进免疫抑制小鼠肿瘤坏死因子（tumor necrosis factor-α,TNF-α）、白细胞介素（interleukin,IL）-1β、干扰素-γ（interferon-γ,IFN-γ）和免疫球蛋白A（immunoglobulin A,IgA）的分泌,调节结肠中免疫相关基因的表达,促进短链脂肪酸（short-chain fatty acids,SCFAs）生成,显著调节Muribaculaceae_unclassified、Muribaculum、Mucispirillum、Helicobacter、Bilophila、Clostridiales等相对丰度（P＜0.05）,逆转CTX诱导的小鼠肠道菌群代谢功能异常,并使其趋于正常。相较于LBP,LB、LB＋LBP还可降低Clostridiales相对丰度,增加有益菌Duncaniella的相对丰度,LB＋LBP还可降低厌氧菌Anaerotruncus的相对丰度。本研究结果将为枸杞精深加工产业中功效针对性健康产品研发和多元化产品开发提供理论依据。     

黑灵芝多糖对免疫抑制小鼠肠道黏膜形态及肠道黏膜免疫的影响

目的：探究黑灵芝多糖对免疫抑制小鼠肠道黏膜形态及肠道黏膜免疫的影响。方法：腹腔注射环磷酰胺构建免疫抑制小鼠模型,将小鼠分为6 组：正常对照组、模型组、阳性对照组以及黑灵芝多糖低、中、高剂量（25、50、100 mg/（kg·d））组,灌胃期间每天记录小鼠体质量并观察其生活状态,空肠组织进行切片病理学观察,酶联免疫吸附实验测定肠道白细胞介素（interlrukin,IL）-6、IL-10、肿瘤坏死因子-α（tumor necrosis factor-α,TNF-α）及干扰素-γ（interferon-γ,IFN-γ）的含量,反转录定量聚合酶链式反应分析T-bet、GATA-3 mRNA相对表达水平。结果：与模型组相比,低、中、高剂量黑灵芝多糖有助于体质量回升,改善肠道黏膜形态结构,提高IL-6、IL-10、TNF-α、IFN-γ细胞因子含量,增强T-bet、GATA-3 mRNA相对表达水平,使Th1/Th2平衡向Th1偏移。结论：黑灵芝多糖可改善免疫抑制小鼠的肠道黏膜形态,调节免疫抑制小鼠的肠道黏膜免疫。     

杨树菇凝集素诱导C57BL/6小鼠肝损伤和活化CD8+T细胞

该研究探究了食用蘑菇杨树菇凝集素AAL(Agrocybe aegerita lectin)对小鼠肝损伤的作用机制。通过小鼠尾静脉注射AAL的方式,根据不同的处理时间分0 h、6 h和9 h三个实验组,分别检测其血清AST(天门冬氨酸氨基转移酶)和ALT(丙氨酸氨基转移酶),对肝脏组织切片进行HE染色,RT-PCR检测肝脏细胞因子表达,流式细胞术对肝脏细胞进行分析。与0h组相比,对小鼠尾静脉注射AAL 6 h和9 h后,小鼠血清ALT和AST显著升高(p0.001);肝组织切片HE染色发现小鼠肝脏呈现明显的肝损伤,可见坏死区和免疫细胞浸润。注射AAL6h后肝脏细胞因子IL-10、IL-33、IL-27、IFN-γ和TNF-α的m RNA表达分别升高到25.14倍、4.62倍、5.14倍、16.51倍和28.10倍。6 h组小鼠肝脏中的T细胞、NKT细胞、CD8+T细胞比例分别由27.92%显著升高到45.95%(p0.05)、2.46%显著升高到6.36%(p0.05)、17%显著升高到27.15%(p0.05)。活化的CD69+T细胞和CD69+CD8+T细胞的比例分别由10.02%显著升高到18.28%(p0.01)和5.96%显著升高到16.20%(p0.01)。结果表明食用菌杨树菇AAL可诱导小鼠肝损伤,活化肝脏CD8+T细胞,显著上调细胞因子IL-10、IL-33、IL-27、IFN-γ和TNF-α的表达,提示食用菌凝集素成分是需要考虑安全使用剂量的。     

川陈皮素对LPS诱导的RAW264.7细胞损伤的保护作用

研究川陈皮素对脂多糖（LPS）刺激的RAW 264.7巨噬细胞损伤的保护作用。MTT法检测川陈皮素的安全用药范围；乳酸脱氢酶试剂盒检测乳酸脱氢酶（LDH）的释放水平；一氧化氮试剂盒检测细胞上清液中一氧化氮（NO）的释放水平；Real-time PCR法检测Toll样受体4（TLR4）、内毒素受体14（CD14）、诱导型一氧化氮合酶（iNOS）、白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）的mRNA表达水平；Western Blot检测核转录因子-κB（NF-κB）的核蛋白表达水平。与正常组比较，LPS刺激组LDH活力和NO的释放量分别升高了4.12倍和3.21倍（p<0.01），iNOS、IL-1β、TNF-α、TLR4和CD14 mRNA表达水平分别升高了2.73倍、4.91倍、10倍、1.84倍和3.53倍（p<0.01）；同时，NF-κB的核表达水平升高了2.50倍（p<0.01）；给予川陈皮素干预后，与LPS刺激组比较，川陈皮素各用药组均能明显降低LDH活力和NO的释放量，减少TLR4、CD14、IL-1β、TNF-α和iNOS的mRNA表达水平和NF-κB的核表达水平（p<0.05或p<0.01）。川陈皮素可减轻LPS刺激RAW 264.7细胞的损伤，机制可能与抑制TLR4-NF-κB信号通路有关。     

蛹虫草多糖调节小鼠巨噬细胞RAW264.7免疫活性的分子机制

多种真菌多糖近年来因其免疫调节活性成为保健食品领域研究的热点。本实验以食药用真菌蛹虫草提取物蛹虫草多糖（Cordyceps militaris polysaccharides,CMP）为研究材料,初步探究CMP对小鼠巨噬细胞RAW264.7免疫活性的调节机制。噻唑蓝实验结果显示CMP无细胞毒性,且100、200 μg/mL CMP可以明显增强RAW264.7细胞活性;中性红法、Griess法和酶联免疫吸附检测结果表明25～200 μg/mL的CMP以剂量依赖方式增强RAW264.7细胞吞噬活性并增加一氧化氮（nitric oxide,NO）和白介素-1β（interleukin-1β,IL-1β）分泌,肿瘤坏死因子-α（tumor necrosis factor-α,TNF-α）的分泌呈先升高后降低的趋势,在100 μg/mL时达到最高值;抑制剂中和实验结果显示当Toll样受体4（Toll-like receptors 4,TLR4）和甘露糖受体（mannose receptor,MR）受到抑制时,CMP诱导的RAW264.7免疫反应显著降低,这表明TLR4和MR均为CMP激活巨噬细胞的受体;此外,Western blot结果显示丝裂原活化蛋白激酶（mitogen-activated protein kinases,MAPK）信号通路也参与CMP诱导的RAW264.7细胞分泌NO、TNF-α和IL-1β。表明TLR4和MR/MAPK信号传导途径在CMP诱导巨噬细胞RAW264.7产生免疫应答时发挥了重要作用。     

Short communication: Pheromonicin-SA affects mRNA expression of toll-like receptors, cytokines, and lactoferrin by Staphylococcus aureus-infected bovine mammary epithelial cells

Pheromonicin-SA (Ph-SA) is a newly developed, engineered multidomain peptide that has a bactericidal effect against Staphylococcus aureus. The objective of this study was to characterize innate immune responses by Staph. aureus-stimulated bovine mammary epithelial cells (BMEC) following treatment with Ph-SA. Primary BMEC from one lactating Holstein cow were isolated and exposed to Staph. aureus for 2 h, and then treated with rifampicin or Ph-SA. Total RNA was isolated from BMEC at 0, 2, 6, 12, and 24 h postinfection, and the mRNA expression of selected genes, including toll-like receptor (TLR)2 and TLR4, IL-1β, IL-6, IL-8, tumor necrosis factor α (TNF-α), and lactoferrin, was quantified by real-time PCR. In the rifampicin group, increases in the expression of mRNA for TNF-α, IL-1β, IL-6, IL-8, and lactoferrin were observed at 6 h postinfection and in the expression of mRNA for TLR2 but not for TLR4 at 12 h postinfection. In the Ph-SA group, increases in the mRNA expression of TLR2, TNF-α, IL-1β, IL-6, IL-8, and lactoferrin were observed at 6 h postinfection, and an increase in TLR4 mRNA expression was observed at 24 h postinfection. At 24 h postinfection, the mRNA expression of TLR4, TNF-α, IL-1β, IL-6, IL-8, and lactoferrin was higher in the Ph-SA group than in the rifampicin group. In conclusion, Ph-SA might promote the expression of mRNA for TLR2, TLR4, the pro-in?ammatory cytokines IL-1, IL-6, and TNF-α, the chemotactic factor IL-8, and lactoferrin in Staph. aureus-infected BMEC. Moreover, Ph-SA may be of value as an antibiotic in promoting innate immune responses by Staph. aureus-infected bovine mammary epithelial cells.     

黑米花色苷矢车菊素-3-O-β-葡萄糖苷对脂多糖诱导的人THP-1单核细胞炎症损伤的保护作用

单核细胞介导的炎症反应在动脉粥样硬化发生和发展过程中起关键作用。花色苷是一种具有多种生物学活性的多酚类黄酮化合物,富含于各种深色的蔬菜、水果及谷类中,其中矢车菊素-3-O-β-葡萄糖苷（cyanidin-3-O-β-glucoside,Cy-3-g）是花色苷中重要的单体,本研究旨在探讨黑米来源的Cy-3-g对脂多糖（lipopolysaccharide,LPS）诱导的人单核白血病细胞（Tohoku hospital pediatrics-1,THP-1）炎症损伤的作用及可能的分子机制。采用乙醇-盐酸溶液浸提、大孔树脂吸附洗脱等步骤得到黑米花色苷粗提物,然后用中压液相层析联合紫外检测提取得到纯化的Cy-3-g（纯度>96.5%）。用不同质量浓度（0、0.10、0.25μg/mL和0.50μg/mL）Cy-3-g与THP-1细胞共孵育4 h,然后加入LPS（质量浓度50 ng/mL）与细胞继续孵育48 h,用酶联免疫吸附试验法测定培养液中白细胞介素-1β(interleukin-1β,IL-1β)、IL-6、IL-8、IL-10和肿瘤坏死因子-α(tumor necrosis fac...     

Serum concentration and mRNA expression in milk somatic cells of toll-like receptor 2, toll-like receptor 4, and cytokines in dairy cows following intramammary inoculation with Escherichia coli

The objective of the current study was to investigate the toll-like receptors (TLR), including the soluble forms sTLR2 and sTLR4, involved in innate immune responses of dairy cows to experimentally induced Escherichia coli mastitis. Six clinically healthy Holstein dairy cows received an intramammary inoculation of E. coli O111:K58 between 63 and 83 d postpartum. Concentrations of sTLR2 and sTLR4, the proinflammatory cytokines IL-6 and tumor necrosis factor-α (TNF-α), and acute phase proteins serum amyloid A (SAA) and haptoglobin (Hp) in blood were measured by ELISA. Furthermore, 10 mL of milk was collected from challenged quarters immediately before inoculation and at 6, 12, 24, 48, and 72 h after inoculation, and mRNA expression of selected genes, including TLR2, TLR4, IL-1β, IL-6, TNF-α, and IL-8, was quantified by real-time PCR. Escherichia coli intramammary infection elicited a decrease in the circulating levels of leukocytes. Rectal temperature was elevated at 6 h postinoculation (PI). Similarly, the serum concentrations of TNF-α, IL-6, and SAA increased at 6 h PI. However, serum concentrations of sTLR2, sTLR4, and Hp did not differ after challenge. The mRNA expression of TLR2, IL-1β, and IL-8 in milk somatic cells increased at 12 h PI, whereas a decreased IL-6 mRNA expression was detected from 6 to 48 h PI. In conclusion, we found that TLR2 mRNA expression increased in milk somatic cells collected from infected quarters of cows challenged with E. coli, whereas the concentrations of sTLR2 and sTLR4 remained unchanged after challenge. Thus, sTLR2 and sTLR4 may protect the host by sequestrating pathogen-associated molecular patterns during E. coli mastitis.     

茯苓多糖通过NF-κB和NLRP3信号通路调节脂多糖引起的焦虑和抑郁样行为

目的：探讨茯苓多糖（Poria cocos polysaccharide,PPS）对脂多糖（LPS）引起的焦虑和抑郁样行为的影响,并分析其机制。方法：BV-2细胞分成对照组、LPS组、LPS+氟西汀组、LPS+PPS(PPS分别为4、8、16μmol/L),处理24 h,二氯二氢荧光素二氢乙酸酯（DCFH-DA）荧光探针检测细胞内活性氧（ROS）水平,Griess法检测培养上清液NO水平,酶联免疫吸附试验（ELISA）检测培养上清液中TNF-α、IL-1β水平,Western blot实验检测CD206、CD16/32、NF-κB p65水平。动物实验将小鼠分为对照组、模型组、LPS+氟西汀组、LPS+PPS低剂量组（20 mg/kg）、LPS+PPS高剂量组（80 mg/kg）,每组10只,测试抑郁样行为,检测海马组织TNF-α、IL-1β、IL-18的浓度,分析海马组织CD206、CD16/32、NF-κB p65、NLRP3、ASC、Cleaved caspase-1蛋白水平。结果：与LPS组比较,4、8、16μmol/L PPS能极显著降低ROS荧光相对强度（P<0.01...     

Immunomodulatory effect of Ganoderma atrum polysaccharide on CT26 tumor-bearing mice

Ganoderma atrum has attracted great attention for its antitumor activity. However, the mechanism remains unclear. A G. atrum polysaccharide (PSG-1) showed pronounced antitumor activity in this study. PSG-1 did not kill CT26 cells directly, but inhibited the proliferation of CT26 cells via the activation of peritoneal macrophages (MΦ). In vivo, PSG-1 significantly suppressed the tumor growth in CT26 tumor-bearing mice. The treatment caused a significant increase in the immune organ index and the phagocytosis of macrophages. The production of TNF-α, IL-1β and nitric oxide also increased. Furthermore, we found that PSG-1 acted on Toll-like receptor (TLR) 4, signaled through p38 MAPK pathway, then activated NF-κB and stimulated TNF-α production. We further found that PSG-1 increased the expression of TLR4 and NF-κB, the degradation of IκBα and the phosphorylation of p38 MAPK. In summary, we have demonstrated that PSG-1 could activate macrophages via TLR4-dependent signaling pathways, improve immunity and inhibit tumor growth.