香栓菌提取物的止咳化痰作用及耐缺氧药理活性研究

本文分别采用浓氨水引咳法、酚红排秘法、常压耐缺氧法、对抗亚硝酸钠中毒性缺氧法,心肌缺血性缺氧法,探讨香栓菌子实体不同极性提取物的止咳化痰及其耐缺氧药理活性。将雄性小鼠随机分组（每组10只）,分别为阴性对照组（NCG）、阳性对照组、香栓菌石油醚提取物低TSPEE(L) 0.025g/kg、中TSPEE(M) 0.05g/kg、高TSPEE(H) 0.1g/kg剂量组、香栓菌水提物低TSWE(L) 0.5g/kg、中TSWE(M) 1.0g/kg、高TSWE(H) 2.0g/kg剂量组。小鼠的检测指标包括咳嗽潜伏期、咳嗽次数、咳嗽抑制率、止咳活性（%）、酚红排泌量、化痰率（%）、常压条件下耐缺氧存活时间、亚硝酸钠中毒存活时间、急性脑缺血性存活时间及血清中SOD、MDA、IL-2、IL-4、IgG含量。结果表明,TSPEE(L)无明显效果;TSPEE(M)具有显著化痰作用（P<0.05）;TSPEE(H)可延长急性脑缺氧存活时间（P<0.05）,并提高SOD、IL-2、IL-4、IgG含量,降低MDA含量（P<0.05）;TSWE(L)可增强咳嗽抑制率并增强止咳活性,同时延长常压条件下缺氧存活时间（P<0.05）;TSWE(M)除中毒性脑缺氧试验外均具有显著性作用（P<0.05）;TSWE(H)可增强咳嗽抑制率、提高止咳活性、增加亚硝酸钠中毒存活时间（P<0.05）,同时增加常压条件和急性脑缺血条件下缺氧存活时间（P<0.01）并且提高SOD、IL-2、IL-4、IgG含量,降低MDA含量（P<0.01）。综上所述,香栓菌子实体石油醚提取物和水提取物具有止咳化痰及耐缺氧作用。     

6种“桑黄”石油醚提取物的体内抗肿瘤活性

对6种＂桑黄＂的石油醚提取物进行了抗肿瘤体内试验,测定其对H22荷瘤小鼠抑瘤率、免疫器官指数、免疫因子含量和生存率的影响。试验结果表明：粗毛纤孔菌、鲍姆木层孔菌、火木层孔菌和瓦宁木层孔菌的石油醚提取物高、低剂量（100,50 mg/kg）以及黑壳目层孔菌石油醚提取物高剂量（100 mg/kg）对肿瘤均具有一定抑制作用,抑瘤率均〉40%;瓦宁木层孔菌石油醚提取物低剂量组（50 mg/kg）的抑瘤效果最佳,为76.82%,其脾指数、胸腺指数均明显高于阳性组（P〈0.01）,IL 2的含量也明显高于对照组和阳性组,能延长小鼠的生存期;椭圆嗜蓝孢孔菌石油醚提取物也具有一定的抑瘤效果,高、低剂量（100,50 mg/kg）抑瘤率分别为39.30%和36.17%。     

裂盖马鞍菌提取物体内抗氧化活性研究

以新疆特色食用菌裂盖马鞍菌为材料,研究裂盖马鞍菌乙醇提取物、乙酸乙酯、氯仿、石油醚萃取物对小鼠体内的抗氧化活性,分高（2g/kg）、中（1g／kg）、低（0．2g／kg）三个剂量对小鼠灌胃给药测定小鼠肝脏和血清中丙二醛（MDA）含量和超氧化物岐化酶（SOD）活力。结果表明,与阴性对照（生理盐水）相比裂盖马鞍菌提取物能明显提高小鼠肝脏和血清中SOD的活力,降低MDA的含量（P〈0．01）;与阳性对照维生素E（VE）（1g/kg）相比乙醇提取物、其他溶剂萃取物高浓度（2g／kg）作用极显著（P〈0．01）;与维生素C（VC,1g／kg）相比乙醇提取物高浓度（2g/kg）作用极显著（P〈0．01）,提示裂盖马鞍菌提取物具有抗氧化活性。     

山野木层孔菌子实体中抑制H22荷瘤小鼠肿瘤的活性成分研究

采用梯度提取法对山野木层孔菌子实体进行提取,得到石油醚层、甲醇层及水层3 种提取物及石油醚层中获得化合物4,6,8(14),22(23)-四烯-3-酮-麦角甾烷,并采用H22 荷瘤小鼠进行体内抗肿瘤活性研究,以抑瘤率、免疫器官指数、免疫因子为指标检测抗肿瘤活性。结果表明,石油醚高剂量组（100mg/kg）、单体化合物中剂量组（7.5mg/kg）抑制率分别为62.21%、57.67%;脾指数、胸腺指数均高于对照组和环磷酰胺（CTX）组,白介素-2（IL-2）的含量明显高于对照组和环磷酰胺（CTX）组（P＜0.01）;肿瘤坏死因子-α（TNF-α）含量明显低于对照组（P＜0.01）。因此认为上述石油醚提取物和单体化合物对H22荷瘤小鼠肿瘤有抑制作用,并且均能改善小鼠的免疫功能。     

哮喘豚鼠IFN-γ／IL-4失衡与IgE水平相关性研究

采用ELISA法、放免法和化学发光法分别观察哮喘豚鼠血清及肺组织匀浆中IFN-γ,IL-4,IgE的含量变化,以及IFN-γ/IL-4与IgE的相关性在哮喘发病机制中的关系和作用。结果表明:哮喘组血清及肺组织一浆中IL-4,IgE含量明显高于对照组(P<0.01),IFN-γ水平明显低于对照组(血清P<0.01;肺组织 P<0.05)。直线相关分析结果表明:IFN-γ,IL-4和IFN-γ/IL-4与IgE呈显著相关性,其中IFN-γ/IL-4与IgE呈显著负相关(血清中 P<0.05;肺组织匀浆中 P<0.01),提示IFN-γ/IL-4的失衡及IgE水平升高在哮喘发病中可能发挥重要作用。     

白囊耙齿菌子实体提取物对小鼠慢性肾小球肾炎的预防与治疗

本文对白囊耙齿菌子实体不同提取物防治慢性肾小球肾炎进行研究,采用腺嘌呤灌胃法建立小鼠慢性肾小球肾炎模型,探究白囊耙齿菌子实体水提取物和醇提取物的高、中、低剂量组对慢性肾小球肾炎的预防作用及不同提取物的高剂量组对慢性肾小球肾炎的治疗作用,并对小鼠体重进行测量,检测尿蛋白(UP)、尿素氮(BUN)、肌酐(Cr)、总胆固醇(TC)、超氧歧化酶(SOD)、一氧化氮(NO)、丙二醛(MDA)、白介素-6(IL-6)、白介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)指标,分析肾、肝、脾、胸腺的病理变化。结果表明,水提取物中剂量(240mg/kg)能显著降低小鼠尿蛋白含量和肾组织中炎症因子含量,使血清生化指标维持正常,并对免疫器官有保护作用;醇提取物高剂量(200mg/kg)对慢性肾小球肾炎有治疗作用,但对免疫器官保护作用较弱。研究表明,白囊耙齿菌子实体水提取物中剂量具有预防慢性肾小球肾炎的作用且效果优于阳性药肾炎康胶囊,具有开发利用潜力,机制有待于进一步研究。     

四种多孔菌子实体粗多糖抗肿瘤活性的比较研究

对粗毛纤孔菌、椭圆嗜蓝孢孔菌、火木层孔菌、木蹄层孔菌4种多孔菌子实体粗多糖成分的含量及其体内抗肿瘤活性进行了比较研究。结果表明粗毛纤孔菌子实体中的粗多糖含量为4.1%,高于其他3种多孔菌;同时,4种多孔菌子实体粗多糖对H22荷瘤小鼠均显示出一定的抗肿瘤活性,除木蹄层孔菌外,其他3种多孔菌给药剂量为500mg/mL和1 000mg/mL时抑瘤率均大于40%,其中粗毛纤孔菌子实体粗多糖抑瘤率最高,低剂量组（500mg/kg）为58.12%,高剂量组（1 000mg/kg）为47.75%。     

毛喉鞘蕊花提取物对大鼠哮喘模型炎症介质的影响及机制研究

为探讨毛喉鞘蕊花提取物(Coleus Forskohlii extract,CFE)对哮喘的治疗作用及机制,采用卵清蛋白(ovalbumin,OVA)结合佐剂氢氧化铝建立大鼠哮喘模型,SPF级SD雄性大鼠60只,随机分为5组,即空白组、模型组、地塞米松组、CFE高剂量组(12.8g/kg)和CFE低剂量组(6.4g/kg),分别用生理盐水、地塞米松及CFE每天灌胃1次,连续14天,灌胃容积为1mL/100g。通过HE染色观察肺组织病理学形态学变化;ELISA法测定大鼠血清(serum)及肺泡灌洗液(BALF)中IFN-γ、IL-4和IL-17A含量变化;Western blot检测MAPK通路相关蛋白的表达变化。HE结果显示,与模型组相比,CFE高、低剂量组能显著减轻OVA刺激后大鼠肺组织病理性损伤,气道轮廓清晰,上皮细胞脱落明显减少;ELISA结果显示,与模型组相比,CFE高、低剂量组大鼠血清及BALF中IL-4、IL-17A的含量明显下降,IFN-γ的含量明显上升(P<0.05或P<0.01);WB结果显示,CFE高剂量组可显著降低大鼠肺组织p-ERK、p-JNK及p-p38蛋白表达(P<0.05或P<0.01)。综上,毛喉鞘蕊花提取物可有效缓解OVA诱导的大鼠哮喘气道炎症反应,其机制与调节MAPK信号传导通路有关。     

特种甘薯TSP-1提取物对环磷酰胺致血小板减少症小鼠的初步研究

ICR小鼠连续3d腹腔注射100mg/kg环磷酰胺,建立血小板减少症模型。将造模成功小鼠分别灌胃6g/kg和2g/kg剂量TSP 1水提取物、6g/kg和2g/kg剂量TSP 1石油醚提取物、2g/kg剂量TSP 1正丁醇提取物和1g/kg剂量TSP 1氯仿提取物,造模前、造模第3天和造模第6天尾部取血测定血小板和白细胞数量。结果表明：2g/kg TSP 1石油醚提取物、1g/kg TSP 1氯仿提取物可以显著升高环磷酰胺模型小鼠的血小板,但各提取物对环磷酰胺所致白细胞减少无治疗作用,提示特种甘薯氯仿与石油醚提取物具有进一步用于治疗血小板减少症的潜在价值。     

灰树花孔菌超微粉体内抗肿瘤作用研究

通过研究肿瘤抑制率、脏器指数、肿瘤切片的观察以及血清中影响因子干扰素-γ（IFN-γ）、白细胞介素-2（IL-2）、血管内皮细胞生长因子（VEGF）、过氧化氢酶（CAT）、谷胱甘肽-过氧化物酶（GSH-PX）和超氧化物歧化酶（SOD）等指标的变化对比分析灰树花孔菌 Grifola frondosa子实体粗粉及灰树花孔菌子实体超微粉对Hep-A-22荷瘤小鼠体内抗肿瘤和免疫调节作用。结果表明,各剂量组分均具有抑制肿瘤生长作用,其中灰树花孔菌超微粉高剂量组（1 500mg/kg）抑瘤效果最佳,与模型组比较有极显著性差异（ P<0.01）。通过HE染色后的肿瘤细胞可观察到坏死面积增大,同时该组血清中IFN-γ、IL-2、VEGF、GSH-PX和SOD含量显著增加（ P<0.01或 P<0.05）,并与抑制肿瘤具有一定相关性。     

香栓菌的驯化栽培及生物学发育特征

为了筛选香栓菌Trametes suaveolens的最佳出菇配方,同时对子实体不同生长阶段的生物学发育特征进行研究。本文用杨树、柳树、秸秆木屑作为培养料设置了8种配比,每种分为对照A、测试组B共计16个配方进行筛选,对不同时期的菌丝发育状况进行显微观察。结果柳树木屑、杨树木屑的配方均能成功栽培出香栓菌子实体,开袋后在90%-95%空气湿度、一定散射光、13-18℃的低温刺激下28d原基发育为菇蕾,继续培养28d后子实体成熟。其中配料为柳树木屑97%、蔗糖1%、石灰1%、石膏1%、含水量65%左右的配方出菇率最高,配料为柳树木屑的配方开袋后子实体长势最佳,采用边缘开袋方式最佳。子实体发育过程中,先分化出白色蜂窝状原基,原基依靠孔状结构吸收营养后,在其上层包被形成黑色菇蕾,同时基部开始形成子实层;菇蕾内部生殖菌丝不断分化和发育,体积逐渐膨大,颜色由深变浅,菌丝分化出担子并产生担孢子,最终形成子实体。     

Therapeutic efficacy of Hypericum perforatum L. extract for mice infected with an influenza A virus

Hypericum perforatum L., a plant used in Chinese herbal medicine, has been proven effective against many viral diseases. In the present study, the therapeutic efficacy of an extract of H.?perforatum (HPE) against influenza A virus (IAV) was investigated in mice. Whether HPE would be a promising agent for influenza treatment was evaluated by measuring the protection rate, mean survival days, lung index, and viral titer, as well as the secretion of IL-6, interleukin-10 (IL-10), tumour necrosis factor-α (TNF-α), and interferon-gamma (IFN-γ) in lung tissue and serum on days 3 and 5 post-infection. The results showed that HPE could reduce the lung index and viral titer of mice infected with IAV, decrease mortality, and prolong the mean survival time. HPE decreased the concentration of IL-6 and TNF-α in lung tissue and serum on day 5 post-infection. In contrast, HPE enhanced the lung and serum levels of IL-10 and IFN-γ on the days 3 and 5 post-infection. Our study indicates that HPE has significant therapeutic efficacy for mice infected with IAV. The possible reasons for these results were concluded to be pertaining to up-regulating the expression of IL-10 and IFN-γ, and down-regulating the secretion of IL-6 and TNF-α in lung and serum.     

应用体内外实验筛选可降低牛乳β-乳球蛋白过敏的乳杆菌

【目的】通过体内外实验评估5种乳杆菌缓解牛乳β-乳球蛋白(BLG)过敏的作用,为今后筛选具有抗过敏活性的乳杆菌提供参考。【方法】首先体外分析5种活的/热致死的乳杆菌促进小鼠原代淋巴细胞分泌细胞因子(CK)IFN-γ和IL-4的水平,随后应用小鼠BLG过敏模型评估这5种乳杆菌抑制过敏的能力。将实验动物随机分为空白组、BLG致敏组和5种活的/热致死乳杆菌组。采用ELISA法检测各组小鼠淋巴细胞分泌Thl/Th2型CK的水平,并测定小鼠血清中总IgE和BLG特异性IgE的含量。【结果】在体外可促进淋巴细胞分泌IFN-γ、抑制IL-4,使其IFN-γ/IL-4比值(代表Thl/Th2细胞平衡)显著高于正常对照组(P<0.05)的乳杆菌,在体内实验中也能有效提高致敏小鼠淋巴细胞的IFN-γ/IL-4分泌率,并显著降低致敏小鼠血清中总IgE和BLG特异性IgE的水平(P<0.05)。相反,在体外的IFN-γ/IL-4比值较低的乳杆菌,不能缓解特异性IgE抗体介导的食物过敏反应。【结论】基于乳杆菌体外刺激小鼠原代淋巴细胞分泌Th1/Th2型CK的结果,可以预测菌株在体内具有可通过纠正Th2占优势的Th1/Th2细胞失衡,下调抗体分泌量,缓解小鼠BLG过敏症状的能力。     

Effects of Citrus unshiu Powder on the Cytokine Balance in Peripheral Blood Mononuclear Cells of Patients with Seasonal Allergic Rhinitis to Pollen

We evaluated the effects of a 50% methanol extract of Citrus unshiu powder (MEC) on cytokines in peripheral blood mononuclear cells (PBMCs) obtained from patients with seasonal allergic rhinitis to cedar pollen. The levels of cytokines, such as TNF-α, IFN-γ, IL-2, IL-4, IL-5, IL-10, IL-12 (p70), IL-13, and GM-CSF, produced by pollen-stimulated PBMC were measured. We found that MEC suppressed pollen-induced TNF-α release and increased IFN-γ release from PBMCs. The results suggest that Citrus unshiu powder has an immunomodulatory effect in vitro and that its use could improve seasonal allergic rhinitis symptoms.     

朱红栓菌提取物抗氧化、抗肿瘤活性评价及相关化学成分分析

采用石油醚、乙酸乙酯、乙醇浸提朱红栓菌 Trametes cinnabarina 子实体干粉,得到不同极性提取物;采用清除DPPH 自由基、羟自由基、超氧阴离子自由基能力,测定提取物的体外抗氧化活性;MTT法检测提取物对人肝癌细胞株HepG2细胞增殖的抑制作用。结果表明,朱红栓菌石油醚、乙酸乙酯、乙醇提取物均具有一定的抗氧化、抗肿瘤活性;各提取物在浓度为4-5mg/mL时,对DPPH自由基、羟自由基和超氧阴离子自由基清除能力大小依次为乙酸乙酯提取物>乙醇提取物>石油醚提取物;乙酸乙酯提取物对3种自由基的最高清除率分别为60.23%、74.49%、63.84%。各提取物对人肝癌细胞株HepG2细胞增殖抑制作用大小依次为乙酸乙酯提取物>乙醇提取物>石油醚提取物;乙酸乙酯提取物的抑制率最高达55.93%。采用硅胶和凝胶等柱色谱方法结合核磁、波谱和质谱等技术对乙酸乙酯提取物的化学组分进行分析,共分离纯化出11种化合物,分别鉴定为：麦角甾醇（1）,邻苯二甲酸二丁酯（2）,对羟基苯甲酸甲酯（3）,麦角甾-7,22,二烯-3-酮（4）,1-[(12E,16E)-12,16-二十碳二烯酰基]-2-[(E,E)-7,11-十八碳二烯酰基]-3-硬脂酰基甘油（5）,cinnabarin（6）,过氧麦角甾醇（7）,尿嘧啶（8）,甘露醇（9）,腺嘌呤核苷（10）,豆甾-7,22-二烯-3β,5α,6β-三醇（11）。除化合物6外均为首次从朱红栓菌子实体中分离得到。研究结果为开发利用朱红栓菌子实体提供了科学依据。     

乳杆菌对牛乳β-乳球蛋白致敏小鼠淋巴细胞分泌Th1/Th2型细胞因子和抗体的体外影响

为了分析乳杆菌对致敏小鼠脾淋巴细胞分泌Th1/Th2细胞因子及抗体的体外影响,用牛乳β-乳球蛋白腹腔注射BALB/c小鼠建立过敏症模型,造模成功后,分离致敏小鼠的脾淋巴细胞并与4种活/死乳杆菌(107 CFU/mL)体外共同孵育,ELISA法检测细胞上清液中细胞因子(IL-12、IFN-γ和IL-4)和抗体(总IgE、β-Lg特异性IgE和总IgG)含量。4种活/死乳杆菌均可体外调节致敏小鼠脾淋巴细胞分泌细胞因子和抗体的水平,特别是热致死的发酵乳杆菌和嗜酸乳杆菌可提高淋巴细胞IL-12和IFN-γ的分泌,抑制IL-4的分泌,使其IFN-γ/IL-4比值(代表Th1/Th2细胞平衡)高于活菌,与空白对照组比较差异显著(P<0.05)。同时,这两株热致死菌还可显著下调细胞上清液中总IgE、特异性IgE和总IgG抗体的浓度(P<0.05)。试验结果表明乳杆菌可提高牛乳β-乳球蛋白致敏小鼠脾淋巴细胞的IFN-γ/IL-4比值,进而纠正Th2占优势的Th1/Th2失衡,下调抗体分泌量,且具有菌株特异性。     

Crude extracts of Caenorhabditis elegans suppress airway inflammation in a murine model of allergic asthma

Epidemiological studies suggest an inverse relationship between helminth infections and allergic disease, and several helminth-derived products have been shown to suppress allergic responses in animals. This study was undertaken to evaluate the effect of a crude extract of Caenorhabditis elegans on allergic airway inflammation in a murine model of asthma. Allergic airway inflammation was induced in BALB/c mice by sensitization with ovalbumin. The effect of the C. elegans crude extract on the development of asthma and on established asthma was evaluated by analyzing airway hyperresponsiveness, serum antibody titers, lung histology and cell counts and cytokine levels in the bronchoalveolar lavage fluid. The role of IFN-γ in the suppression of asthma by the C. elegans crude extract was investigated in IFN-γ knockout and wild-type mice. When mice were sensitized with ovalbumin together with the crude extract of C. elegans, cellular infiltration into the lung was dramatically reduced in comparison with the ovalbumin-treated group. Treatment of mice with the C. elegans crude extract significantly decreased methacholine-induced airway hyperresponsiveness and the total cell counts and levels of IL-4, IL-5 and IL-13 in the bronchoalveolar lavage fluid but increased the levels of IFN-γ and IL-12. Sensitization with the C. elegans crude extract significantly diminished the IgE and IgG1 responses but provoked elevated IgG2a levels. However, the suppressive effect of the C. elegans crude extract was abolished in IFN-γ knockout mice, and the Th2 responses in these mice were as strong as those in wild-type mice sensitized with ovalbumin. The crude extract of C. elegans also suppressed the airway inflammation associated with established asthma. This study provides new insights into immune modulation by the C. elegans crude extract, which suppressed airway inflammation in mice not only during the development of asthma but also after its establishment by skewing allergen-induced Th2 responses to Th1 responses.     

PAS-1, a protein from Ascaris suum,modulates allergic inflammation via IL-10 and IFN-γ, but not IL-12

Helminths and their products have a profound immunomodulatory effect upon the inductive and effector phases of inflammatory responses, including allergy. We have demonstrated that PAS-1, a protein isolated from Ascaris suum worms, has an inhibitory effect on lung allergic inflammation due to its ability to down-regulate eosinophilic inflammation, Th2 cytokine release and IgE antibody production. Here, we investigated the role of IL-12, IFN-γ and IL-10 in the PAS-1-induced inhibitory mechanism using a murine model of asthma. Wild type C57BL/6, IL-12^−/−, IFN-γ^−/− and IL-10^−/− mice were immunized with PAS-1 and/or OVA and challenged with the same antigens intranasally. The suppressive effect of PAS-1 was demonstrated on the cellular influx into airways, with reduction of eosinophil number and eosinophil peroxidase activity in OVA + PAS-1-immunized wild type mice. This effect well correlated with a significant reduction in the levels of IL-4, IL-5, IL-13 and eotaxin in BAL fluid. Levels of IgE and IgG1 antibodies were also impaired in serum from these mice. The inhibitory activity of PAS-1 was also observed in IL-12^−/− mice, but not in IFN-γ^−/− and IL-10^−/− animals. These data show that IFN-γ and IL-10, but not IL-12, play an important role in the PAS-1 modulatory effect.     

Therapeutic potential of Lactobacillus plantarum CJLP133 for house-dust mite-induced dermatitis in NC/Nga mice

Lactobacillus plantarum CJLP133 was isolated from Kimchi, a Korean fermented food, and its potential to improve mouse atopic dermatitis after onset was studied. Dermatitis was developed through house dust-mite extract application onto NC/Nga mice, and then CJLP133 feeding was started. CJLP133 suppressed dermatitis-like skin lesions and decreased high serum IgE levels through balancing between IL-4 and IFN-γ in serum. CJLP133 diminished skin thickening, mast cell accumulation into inflamed site, and lymph node enlargement. In lymph node cells, CJLP133 repressed secretion of T cell cytokines such as IFN-γ, IL-4, IL-5, and IL-10. However, CJLP133 decreased ratios of IFN-γ and IL-5 to IL-10 in lymph node cells, while it did not decrease ratios of IL-4 and IL-5 to IFN-γ. Conclusively, CJLP133 exhibited therapeutic potential for atopic dermatitis in mice through orderly increment of type 1 helper T cell activation and regulatory T cell activation. These results suggest that CJLP133 could treat human atopic dermatitis.