柳茶提取物免疫调节作用的实验研究

对服用柳茶提取物２ｗｋ的小鼠作了免疫机能和形态学方面的实验研究。结果表明：①柳茶的二种提取物均可显著促进淋巴细胞转化；增加正常小鼠抗绵羊红细胞抗体水平；改善腹腔巨噬细胞吞噬功能；水提取物尚能促进ＩＬ－２的产生。②对胸腺重量无影响，但明显增加脾重；光、电镜下见胸腺、脾脏内大量淋巴细胞增生、细胞代谢活跃等改变。提示：柳茶可能是一对免疫功能具多种作用的调节剂。     

戴氏虫草和粉被虫草多糖对巨噬细胞等活性的影响

目的 研究虫草属新种-戴氏虫草菌丝体胞外水溶性多糖（CDP1）和粉被虫草菌丝体多糖（CPP1）在体外对小鼠腹腔巨噬细胞（Mφ）吞噬功能和脾细胞免疫功能的影响。方法 MTT比色法测定淋巴细胞增殖,孔雀绿比色法测定小鼠腹腔巨噬细胞吞噬功能,中性红比色法测定细胞毒淋巴细胞（CTL）活性,结果 CDP1在正常情况下不能促进小鼠腹腔巨噬细胞的吞噬功能,而CPP1却有促进作用;在免疫抑制的情况下,一定浓度的戴氏虫草菌丝体胞外水溶性多糖和粉被虫草多糖能恢复提高吞噬细胞的吞噬功能,而CPP1却有促进作用;在免疫抑制的情况下,一定浓度的戴氏虫草菌丝体细外水溶性多糖和粉被虫草多糖能恢复提高吞噬功能,它们不仅能促进正常脾活化T淋巴细胞的增殖,而且能恢复环磷酰胺和氢化可的松抑制免疫小鼠脾活化T淋巴细胞的增殖,在合适的浓度下能增高小鼠脾CTL活性,同时在一定浓度下能恢复免疫抑制小鼠脾CTL活性。结论 两者具有增强免疫功能的作用。     

淫羊藿多糖对Mφ吞噬功能和Lc自发细胞毒作用的影响

本文观察了淫羊藿多糖对小白鼠腹腔巨噬细胞吞噬功能和脾脏淋巴细胞自发细胞毒作用的影响。淫羊藿多糖(40mg/kg/d×5d,po)组小鼠巨噬细胞吞噬功能比对照增强约1倍;淋巴细胞自发细胞毒作用增强50％(P<0.01)。结果表明淫羊藿多糖对巨噬细胞和淋巴细胞免疫功能具有增强作用。     

锁阳提取物对衰老模型鼠免疫功能及自由基代谢的影响

目的:通过实验探索锁阳多糖、水提取物咀嚼片延缓衰老的效应及其作用机制.方法:提取锁阳多糖、水提取物并制备其咀嚼片;制备衰老动物模型,观察该制剂对衰老模型动物免疫吞噬功能、脾淋巴细胞对刀豆蛋白A(ConA)刺激的增殖活性等的影响;对衰老模型动物脑超氧化物歧化酶(SOD)活性、丙二醛(MDA)、一氧化氮(NO)含量的影响.结果:锁阳咀嚼片治疗组吞噬指数、吞噬百分率、脾脏淋巴细胞转化能力较衰老模型动物升高;锁阳咀嚼片能提高衰老模型动物脑组织SOD活性,降低MDA、NO的含量,且锁阳多糖组在改善自由基水平方面优于锁阳水提物组.结论:锁阳多糖、水提取物咀嚼片可通过改善衰老模型动物免疫功能以及影响脂质过氧化物作用发挥抗衰老作用.     

灵芝孢子粉碱提多糖对小鼠巨噬细胞的免疫调节作用

目的 :研究灵芝孢子粉碱提多糖 (LZSBS)对小鼠巨噬细胞的激活作用。方法 :用灵芝孢子粉碱提多糖刺激体外培养的小鼠巨噬细胞 ,用ELISA法检测巨噬细胞分泌至培养基中的TNF α和IL 1β的含量 ;用Griess法检测培养上清中NO的含量。小鼠巨噬细胞对乳胶颗粒的吞噬率用显微镜计数。结果 :经灵芝孢子粉碱提多糖刺激后 ,小鼠巨噬细胞变大 ,颜色加深 ,并能显著刺激巨噬细胞分泌TNF α和IL 1β ,并产生大量的NO。小鼠巨噬细胞对乳胶颗粒的吞噬功能也明显的增强。结论 :灵芝孢子粉碱提多糖对小鼠巨噬细胞具有明显的激活作用     

淫羊藿多糖对MΦ吞噬功能和Lc自发细胞毒作用的影响

本文观察了淫羊藿多糖对小白鼠腹腔巨吞噬细胞吞噬功能和脾脏淋巴细胞白发细胞毒作用的影响。淫羊藿多糖(40mg/kg/d×5d,PO)组小鼠巨噬细胞吞噬功能比对照增强约1倍(P<0.001);淋巴细胞自发细胞毒作用增强50％(P<0.01)。结果表明淫羊藿多糖对巨噬细胞和淋巴细胞免疫功能具有增强作用。     

蔓越莓水提物促进淋巴细胞增殖并增强其对膀胱癌细胞的杀伤能力

目的研究蔓越莓水提物对淋巴细胞的作用以及经蔓越莓处理的淋巴细胞对膀胱癌的抑制作用。方法运用CCK8细胞活性检测法、BrdU细胞增殖检测法以及细胞杀伤实验等方法研究蔓越莓水提物在体外对淋巴细胞的促增殖作用和淋巴细胞对膀胱癌细胞的抑制杀伤作用。在此基础上,以UPⅡ-SV40T转基因膀胱癌小鼠为模型,喂养蔓越莓水提物,通过HE染色,Ki67免疫组化及细胞杀伤等试验研究蔓越莓水提物对于淋巴细胞的影响及其对膀胱癌的抑制作用。结果本研究用蔓越莓水提物在体外处理小鼠淋巴细胞,发现蔓越莓水提物可以促进淋巴细胞增殖,并提高淋巴细胞对膀胱癌细胞的杀伤作用。体内试验证实,自由摄取含蔓越莓水提物饮水的实验组,小鼠的膀胱质量显著低于普通灭菌水喂养的对照组,同时膀胱癌上皮细胞癌变情况优于对照组。进一步试验发现,实验组血液和脾脏中自然杀伤细胞比例增大,实验组脾脏淋巴细胞对膀胱癌细胞的杀伤作用显著高于对照组。结论蔓越莓水提物促进淋巴细胞的增殖,提高机体杀伤肿瘤细胞的能力,在体内抑制膀胱上皮细胞的癌变和肿瘤的生长,可能对膀胱癌具有预防和治疗作用。     

SAP基因注射干预小鼠SLE样综合征的发生

目的：研究SAP基因注射对SLE发病的干预作用，并进一步探讨SAP发挥作用的可能机制。方法：通过RT-PCR方法克隆SAP基因，构建其真核表达质粒pcDNA3-SAP，观察SAP基因注射对活化淋巴细胞免疫诱导小鼠产生的SLE样综合征的干预作用，以ELISA方法检测抗dsDNA抗体的产生情况，以免疫荧光法检测肾脏免疫复合物沉积；通过巨噬细胞吞噬实验观察SAP与DNA结合后对DNA清除的影响；用增殖实验检测巨噬细胞吞噬SAP结合的DNA后对预致敏淋巴细胞活化的影响。结果：SAP基因注射可有效干预小鼠SLE样综合征的发生，SAP与活化淋巴细胞DNA结合后可明显促进巨噬细胞对DNA的吞噬，并且巨噬细胞吞噬SAP结合的DNA后不引起预致敏淋巴细胞的增殖。结论：提示SAP通过促进DNA等自身抗原的有效清除干预SLE疾病的发生。     

灵芝活性多糖GLIS对正常和荷瘤小鼠骨髓巨噬细胞的激活作用

为研究灵芝活性多糖GLIS对正常和荷瘤小鼠巨噬细胞的激活作用,用灵芝活性多糖GLIS刺激体外培养的正常和荷瘤小鼠的巨噬细胞,检测GLIS刺激后巨噬细胞分泌至培养基中的TNF-α、IL-1β和NO的含量,以及小鼠巨噬细胞对乳胶颗粒的吞噬率的变化,并检测GLIS刺激的巨噬细胞对肿瘤细胞的抑制作用,同时研究灵芝活性多糖GLIS组成的糖和蛋白部分对活性的影响。结果显示经灵芝活性多糖GLIS刺激后,能显著刺激巨噬细胞分泌TNF-α和IL-1β,并产生大量的NO。小鼠巨噬细胞对乳胶颗粒的吞噬功能也明显的增强,且荷瘤小鼠的巨噬细胞对GLIS的敏感度要优于正常小鼠,GLIS中的糖部分对它的活性作用起主要作用。该研究表明,灵芝活性多糖GLIS对正常和荷瘤小鼠巨噬细胞均具有明显的激活作用。     

回阳生肌膏提取物对正常皮肤及慢性溃疡创缘成纤维细胞增殖的影响

目的：比较回阳生肌膏提取物对人正常皮肤成纤维细胞（nHFB）及慢性溃疡创缘成纤维细胞（uHFB）增殖的影响。方法:〖HTSS〗组织块法体外培养nHFB及uHFB；采用 Alarmar blue摄入法观察不同浓度回阳生肌膏水提取物、醇提取物对两种细胞增殖的影响；并采用流式细胞仪检测其水提取物、醇提取物对两种细胞细胞周期的影响。结果:回阳生肌膏水提取物、醇提取物分别作用上述两种细胞24 h后，细胞增殖速度均增加。回阳生肌膏醇提物分别在0.027-0.425 mg/L和0.027-0.213 mg/L范围对nHFB和uHFB的增殖有促进作用（P<0.05或P<0.01）；回阳生肌膏水提物分别在0.075-2.400 mg/L和0.150-1.200 mg/L范围对nHFB和uHFB增殖有明显促进（P<0.05或P<0.01）；流式细胞仪检测结果显示，对于nHFB，水提物作用24 h后 S期比例明显上升（Pμ<0.01），G2/M期比例无明显升高，细胞增殖指数（PrI）明显增加（P<0.01），醇提物作用24 h后S期、G2/M期比例均上升（P<0.05），细胞增殖指数（PrI）明显增加（P<0.01）；而对于uHFB，水提物作用24 h后 S期、G2/M比例上升均不显著，但PrI指数有增加（P<0.05），醇提物作用24 h后 S期比例明显上升（P<0.01），G2/M期比例无明显升高，PrI明显增加（P<0.01）。结论：回阳生肌膏无论水提取物、醇提取物皆可不同程度促进nHFB及uHFB的增殖，其醇提物作用更迅速；这种促增殖作用可能是通过调节细胞周期使更多细胞进入S期而实现的。     

茉莉花、茉莉花茶提取液对部分免疫效应的影响

目的: 观察茉莉花茶提取液和茉莉花提取组分对脾淋巴细胞转化及中性粒细胞增殖活性的影响,为茉莉花茶和茉莉花的应用提供前期研究数据。方法: 采用水提法制作茉莉花茶浸出液,采用石油醚浸泡制备茉莉花脱脑油并进行柱层析分离。采用脾淋巴细胞转化实验与中性粒细胞吞噬实验检测茉莉花茶浸出液的免疫调节效应,另外采用淋巴细胞增殖法(MTT法)测定茉莉花脱脑油和茉莉花提取组分B-Ⅱ的免疫激活作用。结果: (1)茉莉花茶浸出液能明显提高小鼠淋巴细胞转化率,但对中性粒细胞吞噬率无明显影响。(2)MTT证实茉莉花脱脑油和B-Ⅱ能促进小鼠淋巴细胞生长。结论: 茉莉花茶浸出液、茉莉花脱脑油和B-Ⅱ具有一定的免疫促进效应。     

B cell stimulating activity of seaweed extracts

The activity of seaweed extracts on murine and human lymphocytes was studied in vitro. The extracts of some kinds of seaweed, such as Hizikia fusiformis and Meristotheca papulosa, stimulated normal mouse spleen cells to proliferate. The responder cells are B cells, because the response was depleted by the treatment of spleen cells with anti-immunoglobulin (Ig) antibody and complement and being passed through a nylon wool column. This response is not due to lipopolysaccharide (LPS) contamination, because seaweed extracts can stimulate spleen cells of C3H/HeJ mice which are LPS low responders. Seaweed extracts also enhanced Ig production by B cells and tumor necrosis factor (TNF) production by macrophages. Furthermore, seaweed extracts stimulated human lymphocytes to proliferate. All these B cell stimulating activities of seaweed extracts associated with glycoproteins whose molecular weights resided in 100 kD. These results suggest that seaweed extracts have stimulating activity on B cells and macrophages and this ability could be used clinically for the modulation of immune responses.     

红车轴草提取物对小鼠淋巴细胞[Ca2+]i及腹腔巨噬细胞NO分泌和吞噬的影响

目的探讨红车轴草提取物(Trifoliumpratense Leguminosae extract,TLE)体外对小鼠淋巴细胞[Ca2+]i及腹腔巨噬细胞NO分泌和吞噬微球的影响。方法无菌条件下制备小鼠淋巴细胞悬液及小鼠腹腔巨噬细胞悬液;MTT法检测药物对细胞悬液的毒性情况;Fluo-4/AM染色结合流式细胞术分析TLE对小鼠淋巴细胞[Ca2+]i的影响;Griess反应系统检测TLE对脂多糖(LPS)刺激的小鼠腹腔巨噬细胞NO分泌的影响;1μm与2μm直径的荧光微球结合流式细胞术分析TLE对小鼠腹腔巨噬细胞吞噬作用的影响。结果终质量浓度为20、40mg/L的TLE对细胞的毒性小;TLE促进了淋巴细胞的Ca2+内流;TLE抑制了巨噬细胞的NO分泌与吞噬作用,与非TLE组比较P<0.01。结论对淋巴细胞[Ca2+]i及巨噬细胞的NO分泌和吞噬的作用可能是TLE调节小鼠免疫系统的途径。     

积雪草提取物对小鼠脾淋巴细胞凋亡的影响

 目的：分离纯化积雪草活性成份,并观察其对小鼠脾淋巴细胞凋亡的影响。方法：采用乙醇浸提、极性萃取、硅胶柱层析和Sephadex LH-20凝胶柱层析分离纯化积雪草活性成份,通过评价不同阶段积雪草提取物清除1,1-二苯基-2-苦基苯肼自由基（DPPH·）和抑制羟自由基（OH·）能力,筛选积雪草活性成份。采用流式细胞术观察积雪草提取物对小鼠脾淋巴细胞凋亡的影响。结果：(1)积雪草分离纯化后得到提取物B1、B2、B3和B4,其中提取物B4清除DPPH·及抑制OH·的能力最显著。经HPLC分析,积雪草提取物B4主要成分为槲皮素。(2)与对照组相比,积雪草提取物B4作用组小鼠脾淋巴细胞凋亡率显著增加。结论：积雪草提取物B4能诱导小鼠脾淋巴细胞凋亡,其主要成份可能为槲皮素。     

Stimulating activity of Chinese medicinal herbs on human lymphocytes in vitro

The effects of eight kinds of Chinese medicinal herbs (CMH) on human lymphocytes was studied in vitro. The extract of Cinnamomum cassia presl markedly stimulated human lymphocytes to proliferate. Codonopsis pilosula, Oldenlandia diffusa and Rhizoma typhonii weakly stimulated. These extracts enhanced cytotoxic T-lymphocyte (CTL) activity, but failed to enhance natural killer (NK)-cell activity. The extracts of these CMHs have stimulatory effect on immunoglobulin (Ig) production by B-cells and interleukin(IL)-1 production by monocytes. These activities of Cinnamomum cassia presl extract are associated with glycoproteins, whose molecular weight was about 100 KDa. These results suggest that CMH extracts have a stimulating activity on human lymphocytes and these abilities could be used clinically for the treatment of diseases such as cancer.     

中药芦笋促T细胞有丝分裂活性的观察

低浓度芦笋原汁(0.1～1.0％)可明显促进正常小鼠胸腺细胞的增殖,也可刺激正常小鼠脾脏细胞的增殖,但对裸鼠脾脏细胞没有任何作用。由此提示,芦笋具有促T淋巴细胞有丝分裂的活性,而对B淋巴细胞没有作用。与常用的T细胞促有丝分裂素植物血凝素(PHA)及刀豆蛋白A(ConA)不同,芦笋原汁对人、绵羊、豚鼠和鸡的红细胞均无凝集作用。     

Optimization of processing technology of Rhizoma Pinelliae Praeparatum and its anti-tumor effect

BackgroundRhizoma Pinelliae is the dried tuber of Pinellia ternata (Thunb.) Breit. Modern pharmacological studies have shown that Rhizoma Pinelliae has antitussive, antiemetic, glandular secretion inhibiting and antitumor effects.ObjectivesTo optimize the processing technology of Rhizoma Pinelliae Praeparatum, and to study its anti-tumor effect.MethodsOrthogonal design method was applied to analyze the effects of factors such as licorice concentration volume, soaking time and processing temperature on processing technology of Rhizoma Pinelliae Praeparatum; MTT assay and flow cytometry were used to determine the inhibitory effect of Rhizoma Pinelliae Praeparatum on Bel-7402 cells.ResultsDuring the processing of Rhizoma Pinelliae Praeparatum, the size of influence of licorice concentration volume, soaking time and processing temperature on processing results of Rhizoma Pinelliae was: B>C>A in descending order, i.e. soaking time>processing temperature>licorice concentration volume, different concentrations of Rhizoma Pinelliae Praeparatum ethanol extracts could all exert inhibitory effect on the growth and proliferation of Bel-7402 cells, and with the increase of drug concentration and the extension of culture time, the cell proliferation inhibitory effect of Rhizoma Pinelliae Praeparatum ethanol extract became more and more evident. Apoptotic rate of 1.5 mg/ml Rhizoma Pinelliae Praeparatum ethanol extract group reached 13.53%, the difference was extremely significant compared with the control group. In conclusion the factor most influential to the processing technology of Rhizoma Pinelliae Praeparatum was soaking time, followed by processing temperature, the factor least influential was licorice concentration volume.ConclusionRhizoma Pinelliae Praeparatum has inhibitory effect on growth and proliferation of Bel-7402 cells.     

Dithiothreitol-Dependent Antilisterial Activity of Lysates from Normal Macrophages Exposed In Vitro to Culture Fluids from Spleen Cells of BCG-Immunized Mice

The effect of culture fluids from mouse lymphocytes on the antilisterial activity of normal macrophages in vitro was determined. Titers of lysates of macrophages incubated with or without culture fluids from lymphocytes from control mice had titers of 10 to 270 with or without a reducing agent, dithiothreitol. Lysates of macrophages after incubation with culture fluids of spleen lymphocytes from BCG-immunized mice had antilisterial titers with and without dithiothreitol of 197,000 and 810, respectively. This may indicate a dithiothreitol-dependent antilisterial system inherent in activated macrophages of cell-mediated immunity. Lysates of incubated spleen cells and their culture fluids also had slight antilisterial activity.     

Immunomodulating activity of seaweed extract on human lymphocytes in vitro.

Effect of eight kinds of seaweed extract (SWE) on human lymphocytes was studied in vitro. The extracts of Hizikiafusiformis and Meristotheca papulosa (green) markedly stimulated human lymphocytes to proliferate, whereas Eucheuma muricatum and Meristotheca papulosa (red) weakly stimulated proliferation. The responder cells are T cells, because T cells purified by sheep red blood cell (SRBC) rosette-formation were significantly stimulated with SWE, but B cells were not. These extracts enhanced the induction of cytotoxic T lymphocyte (CTL) activity, but failed to enhance natural killer (NK) cell activity. These extracts had a stimulatory effect on immunoglobulin (Ig) production by B cells and tumor necrosis factor (TNF) production by monocytes. The activity of Hizikia fusiformis associated with polysaccharides which were extracted with ethanol and purified by ion-exchange and gel filtration chromatography, whose molecular weight was about 100 kDa. These results suggest that SWE has an immunomodulating activity on human lymphocytes and this ability might be useful for clinical application to treat several diseases such as tumors.     

Effect of anti-kupffer cell serum on phagocytosis and humoral antibody formation

To ascertain the effects of selective impairment of the fixed macrophage compartment on host defense, rabbit anti-rat Kupffer cell serum was raised and tested for its ability to depress phagocytosis and to serve as an immunosuppressant. The ability of the anti-Kupffer cell serum (AKS) to depress intravascular clearance rates of gelatinized RE test lipid emulsion indicated that the antiserum can functionally impair overt phagocytic activity. The phagocytic impairment was manifested by a significantly decreased liver phagocytosis of the lipid emulsion, whereas only a slight decrease in phagocytosis by the spleen was noted. When titered in vitro, AKS was cytotoxic to hepatic and splenic macrophages. Although AKS induced some degree of immunosuppressive activity, it was not as effective as antilymphocytic serum in suppressing the humoral immune response of rats to sheep red blood cells. Selectivity in the cytotoxic activity of AKS was manifested by toxicity for macrophages but not toxicity for thymocytes or splenic lymphocytes. It is suggested that AKS as a specific antimacrophage serum may be useful for assessing the contribution of the reticuloendothelial system to host defense physiology and metabolism.