云芝糖肽对健康人外周血淋巴细胞的体外免疫调节作用

目的探讨云芝糖肽在体外对细胞免疫和体液免疫功能调节的可能机制。方法分离健康人外周血单个核细胞(peripheral blood mononuclear cells,PBMCs),分为PBMCs空白组、云芝糖肽20μL与PBMCs共培养组和云芝糖肽40μL与PBMCs共培养组,3组分别于培养24 h、48 h、72 h采用流式细胞仪检测淋巴细胞亚群的比例变化及T淋巴细胞表面CD3+/HLA-DR+、B淋巴细胞表面CD19+/CD69+表达情况,ELISA方法检测上清液中Ig G水平。结果云芝糖肽与PBMCs共培养组CD4+T细胞比例明显上升(P0.05),CD4+/CD8+细胞比值明显增高(P0.05);云芝糖肽20μL与PBMCs共培养组T细胞表面CD3+/HLA-DR+,72 h时表达水平高于24 h(P0.05);B细胞表面CD19+/CD69+的表达显示从培养24 h到48 h表达升高而在72 h表达减少的趋势;PBMCs与云芝糖肽共培养2组上清Ig G水平均明显高于PBMCs空白组(P均0.05)。结论云芝糖肽在体外能促进CD4+T细胞比例上升及CD8+T细胞比例下降,提高CD4+/CD8+比值,并增加Ig G分泌,具有一定的免疫调节作用。     

The effect of Chinese medicinal herb Zingiberis rhizoma extract on cytokine secretion by human peripheral blood mononuclear cells

The ethanolic extract of the Chinese medicinal herb Zingiberis rhizoma, the rhizome of Zingiber officinale Roscoe (Zingiberaceae), was found to show biphasic effects on secretion of cytokines by human peripheral blood mononuclear cells in vitro. In this study, the augmentative effect of Zingiberis rhizoma extract on cytokine secretion was shown to be time dependent. No significant secretion of cytokine was noted when the reaction time was 1 or 3 h. Secretion of interleukin-1β (IL-1β), interleukin-6 (IL-6), and granulocyte-macrophage colony-stimulating factor (GM-CSF) by the mononuclear cells was markedly increased in the presence of a low concentration of Zingiberis rhizoma extract, varying from 10–30 mg/ml, when the reaction time was 18 or 24 h. A higher concentration of the herbal extract did not show similar or stronger augmentative effect as did low concentration of the herbal extract.     

氟伐他汀对人外周血T淋巴细胞活性的影响

 目的　研究氟伐他汀对人外周血T淋巴细胞活性的影响。方法　采用流式细胞术分析T细胞活化和增殖情况,ELISA法检测细胞因子分泌量,TransAM^TMELISA法测定活化T细胞核因子 (NFAT)的活化状态。结果　氟伐他汀能抑制T细胞表面CD69的表达,显著抑制T细胞增殖,对CD25的表达无影响。能抑制NFAT的活化,显著降低IL-2的产生,对IFN-γ和IL-10的分泌几无影响,但明显诱导IL-4的分泌。结论　氟伐他汀具有免疫抑制作用。     

香加皮羽扇豆烷乙酸酯对人外周血淋巴细胞免疫调节功能的影响

目的 研究香加皮中提取的羽扇豆烷乙酸酯(CPLA)对正常人外周血淋巴细胞和吞噬细胞免疫调节功能的影响.方法 无菌分离人外周血淋巴细胞和巨噬细胞,加入PHA和不同质量浓度的CPLA,MTT法检测其对淋巴细胞增殖及巨噬细.胞对食管癌细胞株ECA-109生长的影响;RT-PCR法检测CPLA对γ-干扰素(INF-γ)mRNA表达的影响;ELISA法检测CPLA作用后淋巴细胞和巨噬细胞培养上清液中肿瘤坏死因子-α(TNF-α)、白细胞介素-2(IL-2)等细胞因子分泌的变化.结果 5～40μg/mL CPLA能明显增强PHA活化的人外周血淋巴细胞的增殖(P<0.01);20、40 μg/mL的CPLA能使淋巴细胞IFN-γ mRNA的表达水平明显升高(P<0.01);用CPLA处理后的巨噬细胞对肿瘤细胞的杀伤功能明显增强(P<0.01),并证实了该作用可能与CPLA促进巨噬细胞分泌TNF-α增多有关;CPLA能促进淋巴细胞产生IL-2和TNF-α,并呈现浓度依赖性(P<0.01).结论 CPLA能增强人外周血淋巴细胞和巨噬细胞的免疫功能,发挥抗肿瘤作用.     

虎杖对人外周血单个核细胞炎症因子基因表达的影响

目的:观察虎杖提取物对人外周血单个核细胞炎症因子基因的影响,以期阐明虎杖提取物抗痛风性关节炎的作用机制。方法:分离与培养人外周血单个核细胞,台盼蓝测细胞活力为95%以上,RT-PCR检测人外周血单个核细胞IL-1βmRNA、IL-8mRNA的表达。结果:虎杖提取物1000mg/L,100mg/L组IL-1βmRNA和IL-8 mRNA表达与空白对照组比较有显著性差异(P﹤0.01)。地塞米松磷酸钠组表达与空白对照组比较有显著性差异(P﹤0.01)。结论:虎杖提取物能抑制人外周血单核细胞炎症因子基因的表达,可能是其治疗急性痛风性关节炎的作用机制之一。     

巴特日七味丸联合金嗓散结丸治疗声带小结的疗效分析

目的:探讨巴特日七味丸和金嗓散结丸联合治疗声带小结的疗效。方法:选择声带小结的患者109例,并将其随机分为治疗组和对照组,治疗组使用巴特日七味丸和金嗓散结丸联合用药治疗声带小结,对照组则单独使用金嗓散结丸。观察和比较两组患者的总有效率、声带小结消失时间、复发率。结果:所有病人经门诊随访半年以上。治疗组总有效率为88.5%,对照组总有效率为71.9%,两组间差异有统计学意义(P0.05)。治疗组声带小结消失时间为(5.8±0.9)周,而对照组为(6.7±1.4)周,两组间差异有统计学意义(P0.05)。复发率比较,治疗组为1.9%,对照组为3.5%,两组间差异无统计学意义(P0.05)。结论:巴特日七味丸和金嗓散结丸联合用药治疗声带小结的临床效果明显优于单独使用金嗓散结丸,既提高了临床疗效,也缩短了治疗时间,值得临床推广应用。     

用寡核苷酸芯片研究血虚小鼠造血相关细胞因子基因表达谱

目的 应用细胞因子寡核苷酸表达谱芯片检测血虚小鼠不同脏器组织内造血相关细胞因子差异表达情况，寻找差异表达基因，为血虚证治疗药物机制研究及治疗药物筛选奠定基础。方法 采用5．5Gy^60Co-γ射线照射Balb／c小鼠，制备血虚模型。在不同时间点提取正常和血虚小鼠不同组织总RNA，反转录成不同荧光标记的cDNA探针，与表达谱芯片进行杂交，对扫描数据进行分析获得血虚小鼠造血相关细胞因子基因的差异表达情况。结果 在各组织中共发现21个差异表达的基因。这些基因的功能大致分为3类：促细胞生长或增殖，免疫调节，诱导血细胞形成或促造血祖细胞形成集落。结论 照射后上述细胞因子基因的下调，使机体的造血功能下降，造成血虚，证明细胞因子间形成造血调节网络，整体调节机体造血。这与中医的全局理论是一致的。实验证明应用芯片技术，从分子水平研究血虚形成的机制是可行的，可为进一步的血虚证治疗药物的筛选提供依据。     

Auraptene from Ferula szowitsiana protects human peripheral lymphocytes against oxidative stress

The antigenotoxicity effects of auraptene on DNA damage in human peripheral lymphocytes were studied using alkaline single cell gel electrophoresis. Auraptene at concentrations of 5, 10, 25, 50, 100, 200 and 400 µM was tested under simultaneous treatment with 25 µM H₂O₂. The data are expressed as % tail DNA and compared with ascorbic acid at concentrations of 25, 50, 100, 200 and 400 µM . Auraptene significantly reduced the genotoxicity of H₂O₂ at concentrations higher than 25 µM (p < 0.001). Interestingly, the antigenotoxicity activity of auraptene was higher than ascorbic acid (p < 0.01), however, at some concentrations (25, 50 and 200 µM ) there was no significant difference between auraptene and ascorbic acid (p > 0.05). It seems that the significant antigenotoxicity effects of auraptene may be due to the prenyl moiety and also the suppression of superoxide anion (O₂^?) generation. This study suggests that the antigenotoxic property of auraptene is of great pharmacological importance and might be beneficial for cancer prevention. Copyright © 2009 John Wiley & Sons, Ltd.     

灵芝提取物对人外周血细胞免疫调节作用的研究

 目的 研究灵芝子实体的水提取物、醇提取物单独及与诱导物一起使用对人外周血吞噬细胞和淋巴细胞的免疫调解作用。方法 以吞噬细胞化学发光、淋巴细胞化学发光和淋巴细胞增殖的方法,观察灵芝提取物的免疫调节作用。结果 单独灵芝提取物不能引起细胞免疫反应,与诱导物一起使用后,低浓度的灵芝提取物对细胞免疫功能有激活作用,高浓度则有免疫抑制作用。结论 与诱导物一起使用后,灵芝水提取物和醇提取物对人外周血吞噬细胞和淋巴细胞均具有双向调节作用。     

The effect of shark liver oil on the tumor infiltrating lymphocytes and cytokine pattern in mice

Ethnopharmacological relevance

Shark Liver Oil (SLO) is a traditional medicine that has been widely used in Scandinavian folk to augment the immune response in some immune-related diseases, especially as an anti-cancer agent.

Aim of the study

The object of this project was to confirm the anti-cancer effect of SLO and the possible involving mechanisms.

Materials and methods

Using delayed-type hypersensitivity (DTH) response in normal mice, the optimal dose for stimulation of cellular immunity was obtained and injected intraperitoneally to the tumor-bearing mice. Cytokine pattern of splenic MNCs was tested by ELISA. The percentage of CD₄⁺ and CD₈⁺ lymphocytes in tumor-infiltrating lymphocytes was determined by flow cytometry. Also the rate of increase in tumor volume measured.

Results

Our findings indicated that SLO highly augments delayed-type hypersensitivity response against sheep Red Blood Cell (sRBC) in mice. Furthermore, intraperitoneal injection of SLO to tumor-bearing mice could increase T-cell infiltration into the tumor and lower the increasing rate of tumor's volume. Also, it changes the cytokine pattern of the splenic Mononuclear cells (MNCs) to Th1.

Conclusion

Increase in IFN-γ (resulting in enhanced cellular immunity) and increase in especially CD₈⁺ lymphocytes accompanied by a decrease in tumor size are among the signs of its anti-tumor effect. Accordingly, we suppose that SLO is a good candidate for further studies in cancer therapy.     

超抗原SEA对人外周血单个核细胞杀伤乳腺癌细胞效力的诱导作用

[目的]研究超抗原葡萄球菌肠毒素A(SEA)对外周血单个核细胞(PBMC)杀伤人乳腺癌细胞(MCF-7)效力的增强作用.[方法]淋巴细胞分离液梯度离心法从人外周血中分离出PBMC,在其中加入终浓度分别为0.01、0.1、1、10mg/L的SEA,与MCF-7在96孔板中共同培养.24、48、72h后用酶标仪检测吸光度值.计算杀伤率.[结果]SEA的促杀伤作用随着时间的增加而增强.SEA与PBMC相互作用72 h后,SEA浓度10 mg/L时,可达到最大杀伤效率.[结论]SEA能够提高PBMC对癌细胞的杀伤作用.     

姜黄素对人淋巴细胞低密度脂蛋白受体表达影响的研究

 目的建立一种简便的直接检测人类低密度脂蛋白受体(lowdensitylipoproteinrecepor,LDL-R)活性并筛选降脂中药的细胞模型:人B淋巴细胞永生化细胞系研究不同浓度姜黄素对人类LDL-R在人淋巴细胞中表达的影响,在细胞和受体水平上探讨姜黄素的作用机制。方法EBV转化技术建立的人B淋巴细胞永生化细胞系;以荧光试剂标记配体法,利用流式细胞仪技术和激光扫描共聚焦显微镜技术研究姜黄素对人淋巴细胞LDL-R表达的影响。结果姜黄素在5～50μmol·L^-1内可以增强人淋巴细胞LDL-R的表达,并且具有明显的量效关系。结论利用荧光试剂标记配体法检测人淋巴细胞LDL-R活性是一个简单有效的方法;姜黄素是一个非常强的LDL-R基因表达促进剂,可能通过增加人淋巴细胞LDL-R的表达起到降血脂和抗动脉粥样硬化的作用。     

四物汤对放射线致血虚证小鼠骨髓Epo和G-CSF基因表达的影响

目的:考察四物汤对放射线致血虚证小鼠的生血作用及对骨髓Epo,G-CSF基因表达的影响,为阐明四物汤补血作用的分子机制提供依据。方法:采用3.5Gy⁶⁰Coγ射线全身1次性照射,制备小鼠血虚证模型;检测外周血象;骨髓集落培养观察并计数CFU-GM,BFU-E,CFU-E,CFUmix;RTPCR检测骨髓Epo和G-CSF基因表达。结果:四物汤显著升高放射线致血虚证小鼠外周血白细胞数量和骨髓CFUGM,BFUE,CFUE,CFUmix数量,上调骨髓Epo和GCSF基因表达。结论:四物汤促进骨髓细胞Epo和G-CSF基因的表达可能是其生血作用的机制之一。     

山楂提取物抑制六价铬对人外周血淋巴细胞致突变作用研究

目的:探讨山楂提取物抑制六价铬的致突变作用.方法:采用人外周血淋巴细胞程序外DNA合成(UDS)试验检测了0.03g、0.015g和0.0075g/ml山楂提取物对重铬酸钾(含Cr6 0.31μg/ml)所致淋巴细胞程序外DNA合成的影响.同时检测山楂提取物与重铬酸钾的加入顺序对UDS的影响.结果:阳性对照(重铬酸钾,含Cr6 0.31μg/ml)组CPM值显著高于阴性对照(蒸馏水)组,P＜0.01.山楂提取物与重铬酸钾同时加入以及两者间隔2h先后加入均显示山楂提取物浓度与CPM值呈显著负相关(P＜0.05),在上述不同加入方法中,山楂提取物的各浓度均显示CPM值显著低于阳性对照.结论:山楂提取物具有抑制Cr6 的致突变作用.     

Hericium erinaceum induces maturation of dendritic cells derived from human peripheral blood monocytes

Hericium erinaceum, a medicinal mushroom, has long been used as a therapeutic due to its immuno‐regulating potentials eliciting anticarcinogenic and antimicrobial efficacies. Since maturation of dendritic cells (DC) is an important process in the initiation and regulation of immune responses, the ability of water‐soluble components from H. erinaceum (WEHE) to regulate DC maturation was investigated. Immature DC were prepared by differentiating human peripheral blood CD14‐positive cells with GM‐CSF and IL‐4. DC were stimulated with WEHE at 2–20 µg/mL for 48 h and subjected to flow cytometric analysis to determine the expression of indicative maturation markers. The endocytic capacity of WEHE‐stimulated DC was examined by a Dextran‐FITC uptake assay. An enzyme‐linked immunosorbent assay was performed to examine the secretion of TNF‐α and IL‐12p40. DC stimulated with WEHE showed representative features upon DC maturation: enhanced expression of CD80, CD83 and CD86, and both MHC class I and II molecules, decreased endocytic capacity of DC, increased expression of CD205, and decreased expression of CD206. However, interestingly, WEHE could not induce the production of TNF‐α and IL‐12p40, whereas lipopolysaccharide substantially increased the production of both cytokines. Collectively, these results suggest that H. erinaceum induces the maturation of human DC, which might reinforce the host innate immune system. Copyright © 2009 John Wiley & Sons, Ltd.     

Effect of Kuibitang on lipopolysaccharide-induced cytokine production in peripheral blood mononuclear cells of chronic fatigue syndrome patients

Kuibitang (KBT) is clinically used to treat patients suffering from chronic fatigue syndrome (CFS) in South Korea. However, its effect has not been investigated experimentally. Recent reports have shown that CFS patients display an altered cytokine production. We examined the effect of KBT on lipopolysaccharide (LPS)-induced various cytokines production in peripheral blood mononuclear cells (PBMC) of CFS patients and healthy controls. KBT (1 mg/ml) significantly inhibited LPS-induced tumor necrosis factor-, interleukin-10, and transforming growth factor-β1 production in PBMC of CFS patients. However, LPS-induced interferon-γ production was significantly increased by KBT (0.01 mg/ml). These results provide evidence of a novel activity of the KBT that regulate cytokines production related with CFS.     

环孢素对心脏移植大鼠外周血淋巴细胞多基因mRNA表达的影响

 目的 对大鼠心脏移植前后不同时间点外周血淋巴细胞mRNA差异表达的研究,探讨环孢素对免疫排斥反应过程中基因表达的影响.方法 心脏移植前后以15 mg·kg^-1·d^-1环孢素灌胃的大鼠,分别在移植前、移植后24,72 h,7,10,12d取血,提取总RNA,用mRNA差异显示法找出差异表达条带,克隆测序,用灰度扫描法计算差异表达基因的比密度。结果 对照组移植心脏存活率于移植后的10d内降至为0%,而此阶段环孢素组的移植心脏存活率始终保持在50％.移植后第10天,对照组与凋亡信号传导和线粒体蛋白构象重构与蛋白转运相关基因Ribosomal—12S和S25,MHC—Ⅲ(Hst-70)的表达水平显著高于移植前,与移植心脏存活率最低点同步.其2,3-bisphosphoglycerate和gig18基因表达水平的变化无统计学意义。环孢素组与细胞凋亡有关的gig18和Thbsl,以及T细胞受体(TCR)等基因的表达水平于移植后10 d内持续低于移植前,2,3-bisphosphoglycerate基因的表达水平于移植后24 h降低,只有钠通道β-1基因的表达水平于移植后10 d内高于移植前。结论 环孢素可负调心脏移植后大鼠外周血淋巴细胞凋亡相关基因gig18和Thbsl,正调钠通道基因的转录。     

紫茜胶囊对ITP模型小鼠外周血细胞因子IL-2及其受体的影响

目的:观察紫茜胶囊对特发性血小板减少性紫癜(ITP)模型小鼠外周血细胞因子IL-2及其受体的变化,以及细胞因子IL-2与ITP发病的关系.方法:采用放免法测定IL-2及其受体.结果:ITP模型小鼠外周血细胞因子IL-2水平降低,IL-2受体水平升高,与模型组相比有明显差异(P<0.05).结论:紫茜胶囊对ITP模型小鼠外周血细胞因子具有明显的调节作用.