天盾液对免疫低下小鼠免疫功能的影响

目的研究天盾液对免疫低下模型小鼠免疫功能的调节作用。方法小鼠ip环磷酰胺,造成免疫功能低下的模型,观察天盾液对小鼠免疫器官重量、碳粒廓清能力、迟发型过敏反应、血清半数溶血素值(HC50)及脾细胞形成抗体功能的影响,盐酸左旋咪唑作为阳性对照。结果天盾液能显著增加免疫低下小鼠脾脏的重量,明显增强巨噬细胞的吞噬功能,提高绵羊红细胞所致的迟发型过敏反应和HC50水平;提升脾细胞分泌抗体的能力。结论天盾液能明显增强免疫功能低下小鼠的免疫功能。     

天螺霜抗乙型肝炎病毒作用的研究

目的观察天螺霜体内、体外抗乙型肝炎病毒作用。方法采用实时荧光定量PCR(FQ-PCR)体外抑制试验及鸭乙型肝炎动物模型体内试验观察天螺霜对乙型肝炎病毒复制过程的影响。结果FQ-PCR试验显示,蒸馏水组与天螺霜组DNA拷贝中位数分别为6.9×108/ml与2.6×104/ml,差异有统计学意义(u=2.76,P<0.01)。体内试验25mg.kg-1.d-1和100mg.kg-1.d-1剂量组在用药5d、10d及停药3d时均能显著降低鸭乙肝病毒(DHBV)感染模型动物血清DHBV-DNA水平(P<0.05或P<0.01),两剂量组用药5d、10d及停药3d时DHBV-DNA抑制率分别为51.84%,55.32%,36.88%和81.08%,87.10%,61.80%。结论天螺霜体内、体外均具有抗乙型肝炎病毒复制的生物学作用。     

猫爪草多糖对环磷酰胺致小鼠免疫低下模型免疫功能的影响

目的 探讨猫爪草多糖对环磷酰胺致小鼠免疫抑制模型免疫功能的作用特点。方法 应用环磷酰胺建立免疫抑制模型,观察大、中、小剂量猫爪草多糖对模型小鼠腹腔巨噬细胞吞噬功能、溶血素和溶血空斑形成以及淋巴细胞转化的影响。结果 大、中剂量猫爪草多糖可显著提高免疫抑制小鼠吞噬百分率和吞噬指数;中剂量猫爪草多糖和香菇多糖组可显著促进溶血素的形成;大、中、小剂量猫爪草多糖和香菇多糖均可显著促进溶血空斑的形成、促进淋巴细胞转化。结论 猫爪草多糖可明显改善环磷酰胺致免疫抑制小鼠免疫功能。     

玄参多糖对正常生理小鼠和免疫低下状态小鼠免疫功能的影响

目的:考察玄参多糖对正常生理小鼠和免疫低下状态小鼠免疫功能的影响。方法:将80只小鼠随机分为正常组,模型组,治疗高、中、低剂量组和生理高、中、低剂量组,每组10只。正常组和模型组小鼠ig蒸馏水(10 mL/kg),治疗高、中、低剂量组和生理高、中、低剂量组均按0.16、0.08、0.04 g/kg ig给药,每天1次,连续7 d;于给药第3天开始,模型组和治疗高、中、低剂量组小鼠分别ip环磷酰胺(100 mg/kg),连续4 d,复制免疫低下模型。采用碳廓清实验测定小鼠碳廓清指数和脏器(胸、脾)指数。另取80只小鼠分组、给药同上,采用绵羊红细胞致敏后测定其血清半数溶血值(HC_(50))。再另取80只小鼠分组、给药同上,采用1%二硝基氟苯法诱导小鼠迟发型变态反应测定其耳肿胀度,测定其血清中白细胞介素(IL)-2、IL-4、血清免疫球蛋白G(IgG)、免疫球蛋白M(IgM)、γ-干扰素(IFN-γ)含量,并考察脾淋巴细胞的体外增殖情况。结果:高剂量玄参多糖可促进两种免疫状态下小鼠免疫器官及碳廓清指数的增长(P<0.05);增强小鼠迟发型变态反应的强度(HC_(50)值升高)(P<0.01)及改善模型小鼠血清溶血素含量的降低(P<0.01);促进脾淋巴细胞增殖(P<0.05)及升高血清中IL-2、IL-4、IgG、IgM、IFN-γ含量(P<0.05或P<0.01)。结论:玄参多糖可增强正常生理和环磷酰胺所致免疫低下两种状态下小鼠的免疫功能。     

小叶黑柴胡总多糖对小鼠免疫功能的影响

目的研究小叶黑柴胡总多糖对小鼠免疫功能的影响。方法将小鼠随机分为空白对照组、阳性药对照组、小叶黑柴胡总多糖2个剂量（40、20mg·kg^-1）组,灌胃给药。检测药物对绵羊红细胞诱导的小鼠迟发型变态反应的影响;测定小鼠T、B淋巴细胞转化功能;检测药物对小鼠血清溶血素水平的影响;中性红比色法测定小鼠腹腔巨噬细胞胞饮功能,硝酸还原酶法测定巨噬细胞培养上清液中一氧化氮水平。结果小叶黑柴胡总多糖40mg·kg^-1显著抑制迟发型变态反应（P〈0．05）;小叶黑柴胡总多糖20mg·kg^-1显著增强腹腔巨噬细胞胞饮能力（P〈0．05）;显著升高血清溶血素水平（P〈0．001）。小叶黑柴胡总多糖刺激淋巴细胞增殖,并协同伴刀豆球蛋白A促进小鼠T淋巴细胞增殖（P〈0．05）。结论小叶黑柴胡总多糖对小鼠免疫功能有调节作用。     

海参提取物对免疫功能低下模型小鼠免疫功能的调节作用

目的:研究海参提取物对免疫功能低下模型小鼠免疫功能的调节作用。方法:50只ICR小鼠随机均分为正常对照(等容生理氯化钠溶液)组、模型(等容生理氯化钠溶液)组、左旋咪唑(40 mg/kg)组与海参提取物高、低剂量[500、250 mg(生药)/kg]组,灌胃给药,每天1次,连续10 d。给药7 d后,腹腔注射环磷酰胺(40 mg/kg),每天1次,连续3 d以复制小鼠免疫功能低下模型。小鼠尾静脉注射0.5%刚果红溶液(0.1 ml/10 g),采血,分光光度计测定其光密度以评价单核巨噬细胞吞噬能力;细胞计数仪读取白细胞数;酶联免疫吸附(ELISA)法检测血清免疫球蛋白(Ig)G水平;称定小鼠胸腺、脾脏与体质量以计算免疫脏器指数;2(-2-甲氧基-4-硝基苯基)-3-(4-硝基苯基)-5-(2,4-二磺酸苯)-2H-四唑单钠盐(CCK-8)法检测淋巴细胞增殖能力。结果:与正常对照组比较,模型组小鼠吞噬指数(κ)、校正吞噬指数(α)差异无统计学意义(P>0.05);白细胞数减少,血清Ig G含量减少,胸腺指数、脾脏指数降低,脂多糖(LPS)刺激增殖率、刀豆蛋白A(Con A)刺激增殖率降低,差异均有统计学意义(P<0.01)。与模型组比较,海参提取物高、低剂量组小鼠α升高,血清Ig G含量增加;海参提取物高剂量组κ升高、白细胞数增加,差异均有统计学意义(P<0.01或P<0.05);海参提取物高、低剂量组胸腺指数、脾脏指数、LPS刺激增殖率、Con A刺激增殖率差异无统计学意义(P>0.05)。结论:海参提取物对化疗药物环磷酰胺所致小鼠免疫功能低下具有一定的调节改善作用。该研究为海参调节免疫功能的临床应用提供了实验依据。     

尼美舒利对正常小鼠免疫功能的影响

目的：研究尼美舒利对免疫功能的影响。方法：采用碳粒廓清法、二硝基氟苯诱导迟发型变态反应试验法、抗体形成测定法及免疫器官重量法，评价尼美舒利对正常小鼠免疫功能的作用。结果：尼美舒利能增强二硝基氟苯诱导的迟发型变态反应，并且提高抗体水平增加脾脏重量。结论：尼美舒利能增强细胞免疫和体液的免疫功能。     

六堡茶对小鼠免疫功能的影响

目的探讨六堡茶对实验小鼠免疫功能的影响。方法采用KM小鼠,分为对照组和不同剂量组,分别经口给予低、中、高剂量的六堡茶30d后,观察不同剂量的六堡茶对小鼠迟发型变态反应(DTH)、溶血空斑形成的影响。结果六堡茶中、高剂量可提高小鼠溶血空斑数,但对小鼠DTH无明显影响。结论六堡茶能增强小鼠体液免疫功能。     

淫羊藿多糖对免疫功能低下小鼠免疫功能的影响

采用环磷酰胺所致免疫低下小鼠及荷瘤 (S180 )小鼠观察了淫羊藿多糖 (EPS)对免疫功能的影响 .结果表明 :腹腔注射环磷酰胺 80mg/kg ,可以使正常小鼠的胸腺指数、血清溶血素水平、空斑形成细胞溶血能力和外周血中WBC总数下降 .腹腔注射EPS 5 0、2 5mg/kg× 7,可使脾指数上升 ,但对胸腺指数无明显影响 ;5 0、2 5、12 5mg/kg× 7,使下降的溶血素值及空斑形成细胞的溶血能力回升 ;2 5、12 5mg/kg× 7,使下降的白细胞总数上升 .小鼠荷瘤后免疫功能低下 ,表现为胸腺指数下降、血清溶血素水平、空斑形成细胞的溶血能力及迟发性超敏反应能力降低 .皮下注射EPS5 0mg/kg× 8,可对抗荷瘤引起的胸腺指数下降 ;5 0、2 5mg/kg使荷瘤小鼠的脾指数升高 ;5 0、2 5mg/kg× 8,可以使荷瘤小鼠的血清溶血素、空斑形成细胞的溶血能力及迟发性超敏反应能力有所提高 .     

小分子阿胶对小鼠免疫功能的影响

目的 研究小分子阿胶对小鼠免疫功能的调节作用。方法 将ICR小鼠随机分为对照组、模型组、生血丸3.00 g/kg组和小分子阿胶3.00、1.50、0.75 g/kg组,ig给药14 d,ip环磷酰胺或氢化可的松建立免疫低下模型,用鸡红细胞诱导小鼠溶血素,测定小鼠血清溶血素水平,观察小分子阿胶对体液免疫的影响;采用二硝基氯苯诱导小鼠耳廓肿胀,检测小鼠耳廓肿胀度,观察小分子阿胶对细胞免疫的影响;利用流式细胞术对小鼠淋巴细胞亚群进行分析,观察小分子阿胶对小鼠淋巴细胞分型的影响。结果 与模型组比较,小分子阿胶3.00、1.50、0.75 g/kg显著提高免疫功能低下小鼠血清溶血素含量（P<0.01）;3.00、1.50 g/kg显著提高小鼠耳廓肿胀度（P<0.01、0.05）;3.00 g/kg显著提高小鼠T淋巴细胞（CD3⁺）、辅助性T细胞和迟发超敏性T细胞（CD3⁺CD4⁺）占淋巴细胞的比例（P<0.01）。结论 小分子阿胶能提高小鼠体液免疫和细胞免疫功能,对免疫功能有正向调节作用。     

绞股蓝总苷对免疫低下小鼠非特异性免疫功能的影响

目的 探讨绞股蓝总苷免疫增强的药理作用，观察剂量效果关系。方法 用绞股蓝总苷作为免疫增强剂治疗免疫低下小鼠模型，通过碳粒廓清实验观察非特异性免疫功能的变化。小鼠随机分为六组，模型组皮下注射环磷酰胺40mg／kg，制造免疫低下模型，与各对照组和药物高、中、低剂量组分别作碳粒廓清实验，观察各组非特异性免疫功能变化。结果 模型小鼠廓清指数明显下降，阳性对照组，绞股蓝高、中剂量组非特异性免疫功能指标明显增强，作用强度相似。绞股蓝总苷的作用呈剂量依赖性。结论 绞股蓝总苷能非常明显地增强非特异性免疫功能。     

Influence of MHC background on the antibody response to detergent enzymes in the mouse intranasal test.

The mouse intranasal test (MINT) was developed to assess the immunogenic potential of detergent enzymes. The BDF1 mouse (H-2(b/d)), a cross of C57Bl/6 (H-2(b)) x DBA/2 (H-2(d)) has been used for most of the development work. Preliminary data in the CB6F1(H-2(d/b)), a cross of Balb/c (H-2(d)) x C57Bl/6 showed that this strain was similar to the BDF1 in its response to enzymes. These data also showed that the parental strains responded differently to the enzymes. To understand better the influence the major histocompatibility complex (MHC) background has on immune responses to enzymes, 3 different enzymes were tested in 4 inbred strains (C57Bl/6, DBA/2, Balb/c, and C57Bl/10), 2 hybrid strains (BDF1 and CB6F1), and 2 congenic strains (Balb.B10 and B10.D2). BDF1 mice rank enzymes the same as the guinea pig, which in turn correlates with sensitization in occupationally exposed humans. The ranking is based upon the dose of enzyme needed to give one-half maximal IgG1 antibody response (ED(50)) where Termamyl is more potent than Alcalase, which is equipotent to Savinase. The H-2(d) strains ranked the enzymes the same as the BDF1 but generated ED(50)s for the proteases that were one order of magnitude greater than the BDF1 ED(50)s. The response to Termamyl was the same in the two F1 strains and the H-2(d) strains. The H-2(b) strains did not rank the enzymes the same as BDF1 but the ED(50)s for the proteases were similar to the ED(50)s in the F1 strains. The response to Termamyl in the H-2(b) strains was lower than the response in the F1 and H-2(d) strains. Initial data show that the inbred strains will make enzyme-specific IgE antibody to high doses of enzyme with DBA/2 > Balb/c > C57Bl/6 in terms of the robustness of the response. The IgG1 responses in the congenic strains were similar to the responses in the H-2 matched strains. In addition, the antibody response to enzymes was consistent regardless of the source of BDF1 mice. The responses to these enzymes are clearly MHC linked with a role for Class II I-E molecules indicated. The current data strongly support the use of the F1 hybrid as an appropriate strain for evaluating allergic responses to enzymes.     

重型乙型肝炎血清HBV DNA水平与肝功能及免疫指标的关系研究

目的 探讨重型乙型肝炎患者血清HBV DNA水平与肝损害程度以及血清免疫指标之间的关系.方法 应用荧光定量聚合酶链反应法和酶联免疫吸附法（ELISA）分别检测197例乙肝患者血清HBVDNA水平与乙型肝炎病毒血清标志物（HBVM）.结果 重型乙型肝炎患者血清HBV DNA水平显著低于慢乙肝重度,慢乙肝轻、中度及急性乙肝（P＜0.01）;重型乙型肝炎患者HBeAg阴性率最高（P＜0.05）;重型乙肝患者血清HBV DNA水平与ALT、TBil无相关性,与C3、C4呈显著正相关（P＜0.01）,与PT、IgG、IgA、IgM呈显著负相关（P＜0.05～0.01）;重型乙型肝炎死亡患者血清HBV DNA水平显著低于存活者（P＜0.05）.结论 重型乙型肝炎血清HBV DNA复制水平低,HBeAg阴性率高,与病毒变异有关;重型乙型肝炎血清HBV DNA水平与肝功能及免疫指标有良好相关性;定量检测重型乙型肝炎患者血清HBV DNA对判断病情危重程度、指导抗病毒治疗和估计预后有重要意义.     

Effects of microsomal enzyme inducers in vivo and inhibitors in vitro on the covalent binding of benzo[a]pyrene metabolites to DNA catalyzed by liver microsomes from genetically responsive and nonresponsive mice.

The in vitro DNA binding of benzo[a]pyrene metabolites generated by mouse liver microsomes can be resolved into at least nine distinct peaks by elution of a Sephadex LH20 column with a water-methanol gradient. These peaks, representing metabolite-nucleoside complexes, are named A (most polar) through I (least polar). 3-Methylcholanthrene, 2,3,7,8-tetrachlorodibenzo-p-dioxin, phenobarbital, Aroclor 1254, pregnenolone-16α-carbonitrile, or ethanol was administered in vivo to genetically “responsive” C57BL/6N or “nonresponsive” DBA/2N mice, in an attempt to understand and identify increases or decreases in reactive BP intermediates that bind to DNA. Rises or falls in these peaks are also noted when liver microsomes from control or 3-methylcholanthrene-treated C57BL/6N or DBA/2N mice were incubated in vitro with [³H]benzo[a]pyrene and microsomal enzyme inhibitors such as α-naphthoflavone, metyrapone or cyclohexene oxide. All of our interpretations concerning the binding of metabolites to DNA are consistent with non-K-region oxygenation of benzo[a]pyrene being mediated predominantly by cytochrome(s) P₁-450 and K-region oxygenation of benzo[a]pyrene being catalysed predominantly by form(s) of P-450 other than P₁-450. All of the biological perturbations are consistent with the following assignments. The major reactive intermediate of benzo[a]pyrene contributing to each peak is suggested to be: peaks A and C, an unknown dihydrodiol oxide; peaks B, D, F and I, quinones oxygenated further (or quinone-derived free radicals); peak E, both cis- and tans-7,8-diol-9,10-epoxides; peak F′, the 7.8-oxide; peak G, the 4.5-oxide; and peak H, an unknown phenol oxide. The DNA nucleosides are not identified in this study. Of the ten peaks listed here, it is of interest that the major metabolite(s) contributing to eight of the peaks (all except peaks F' and G) involve(s) more than a single mono-oxygenation by forms of cytochrome P-450. All peaks, with the exception of peak G, appear to be predominantly associated with benzo[a]pyrene metabolism mediated by P₁-450 and, therefore, controlled by the Ah locus. The use of these microsomal enzyme inducers or inhibitors—combined with the underlying genetic predisposition of the individual, tissue, or cell culture system under study—demonstrates that the balance between P-450 and epoxide hydrase, and the ratio of each form of P-450 to the other forms of P-450, can influence markedly the quantity and quality of reactive intermediates of benzo[a]pyrene that bind to DNA.     

Contribution of CYP2C9, CYP2A6, and CYP2B6 to valproic acid metabolism in hepatic microsomes from individuals with the CYP2C9*1/*1 genotype.

The present study investigated the role of specific human cytochrome P450 (CYP) enzymes in the in vitro metabolism of valproic acid (VPA) by a complementary approach that used individual cDNA-expressed CYP enzymes, chemical inhibitors of specific CYP enzymes, CYP-specific inhibitory monoclonal antibodies (MAbs), individual human hepatic microsomes, and correlational analysis. cDNA-expressed CYP2C9*1, CYP2A6, and CYP2B6 were the most active catalysts of 4-ene-VPA, 4-OH-VPA, and 5-OH-VPA formation. The extent of 4-OH-VPA and 5-OH-VPA formation by CYP1A1, CYP1A2, CYP1B1, CYP2C8, CYP2C19, CYP2D6, CYP2E1, CYP4A11, CYP4F2, CYP4F3A, and CYP4F3B was only 1-8% of the levels by CYP2C9*1. CYP2A6 was the most active in catalyzing VPA 3-hydroxylation, whereas CYP1A1, CYP2B6, CYP4F2, and CYP4F3B were less active. Correlational analyses of VPA metabolism with CYP enzyme-selective activities suggested a potential role for hepatic microsomal CYP2A6 and CYP2C9. Chemical inhibition experiments with coumarin (CYP2A6 inhibitor), triethylenethiophosphoramide (CYP2B6 inhibitor), and sulfaphenazole (CYP2C9 inhibitor) and immunoinhibition experiments (including combinatorial analysis) with MAb-2A6, MAb-2B6, and MAb-2C9 indicated that the CYP2C9 inhibitors reduced the formation of 4-ene-VPA, 4-OH-VPA, and 5-OH-VPA by 75-80% in a panel of hepatic microsomes from donors with the CYP2C9*1/*1 genotype, whereas the CYP2A6 and CYP2B6 inhibitors had a small effect. Only the CYP2A6 inhibitors reduced VPA 3-hydroxylation (by approximately 50%). The extent of inhibition correlated with the catalytic capacity of these enzymes in each microsome sample. Overall, our novel findings indicate that in human hepatic microsomes, CYP2C9*1 is the predominant catalyst in the formation of 4-ene-VPA, 4-OH-VPA, and 5-OH-VPA, whereas CYP2A6 contributes partially to 3-OH-VPA formation.     

Signal transduction pathways implicated in the decrease in CYP1A1, 1A2 and 3A6 activity produced by serum from rabbits and humans with an inflammatory reaction

Incubation of serum from rabbits with a turpentine-induced inflammatory reaction and from humans with an upper respiratory viral infection with hepatocytes from rabbits with a turpentine-induced inflammatory reaction for 4h reduces total cytochrome P450 content and activity of cytochrome P450 isoforms CYP1A1/1A2 and 3A6 without affecting the expression of these proteins. To document the signal transduction pathways implicated in the decrease in CYP1A1/1A2 and 3A6 activity, hepatocytes from rabbits with a turpentine-induced inflammatory reaction were incubated with serum from rabbits with a turpentine-induced inflammatory reaction, serum from individuals with a viral infection and interleukin-6 for 4h in presence of inhibitors of protein kinases. The sera-induced decrease in CYP1A1/1A2 and 3A6 activity was partially prevented by the inhibition of Janus-associated protein tyrosine kinase, double-stranded RNA-dependent protein kinase, protein kinase C, and p42/44 mitogen-activated protein kinase. The serum from rabbits with a turpentine-induced inflammatory reaction increased the phosphorylation of Erk1/2, effect prevented by PD98059 but not by bis-indolylmaleimide, a specific inhibitor of protein kinase C. The results demonstrated that the decrease in total cytochrome P450 content and in CYP1A1/1A2 and 3A6 activity by sera and interleukin-6 involves the activation of protein tyrosine kinases, p42/44 mitogen-activated protein kinase and protein kinase C. Indirect evidence supported that nitric oxide is implicated in the decrease in activity of these enzymes.