含白花蛇舌草血清对大鼠淋巴细胞和混合培养淋巴细胞增殖影响的实验研究

目的:研究含白花蛇舌草(Spreading Hedvotis Herb,简称SH)血清对植物血凝素(PHA)诱导大鼠脾淋巴细胞增殖、混合淋巴细胞培养(简称MLR)淋巴细胞增殖及对淋巴细胞分泌白细胞介素-2(IL-2)和γ-干扰素(IFN-γ)的影响。方法:大鼠20只,随机分为5组,每组4只,分为SH低剂量(4 g/kg)、中剂量(16 g/kg)、高剂量(24 g/kg)组,含环孢素A(CsA)血清组,按相应药物灌胃,空白血清组用等量生理盐水灌胃。MTT法测定PHA诱导的大鼠脾淋巴细胞和MLR淋巴细胞增殖,ELISA法测定PHA诱导淋巴细胞分泌IL-2、IFN-γ含量。结果:SH高剂量组可抑制PHA诱导大鼠脾淋巴细胞增殖,SH高剂量组、含CsA血清组与空白血清组比较,差异均有显著性意义(P0.05)。双向MLR体系中,各剂量组含SH血清均可抑制淋巴细胞增殖,其中SH高剂量组与空白血清组比较,差异有显著性意义(P0.05);在单向MLR体系中,各剂量组含SH血清可调节淋巴细胞增殖。SH中剂量组、SH高剂量组血清抑制脾淋巴细胞增殖,与空白血清组比较,差异均有显著性意义(P0.05)。含CsA血清组血清可抑制单向、双向MLR体系中淋巴细胞增殖,与空白血清组比较,差异均有显著性意义(P0.05)。SH中剂量组、SH高剂量组、含CsA血清组血清均可显著抑制大鼠脾淋巴细胞分泌IL-2、IFN-γ含量,与空白血清组比较,差异均有显著性意义(P0.05)。结论:SH能抑制淋巴细胞和混合培养淋巴细胞增殖,其作用机制可能与抑制淋巴细胞分泌IL-2、IFN-γ有关。     

拯阳汤的含药血清对阳虚小鼠免疫功能的影响

目的：采用血清药理学实验方法，观察拯阳汤的含药血清对阳虚小鼠免疫功能的影响。方法：采用氢化可的松（HC）阳虚小鼠模型，用MTT法检测腹腔巨噬细胞（pMФ）的活化作用及其分泌IL-1的活性，脾脏T淋巴细胞的增殖活性及其分泌的IL-2的活性，并用原位末端标记方法（TUNEL）检测胸腺细胞凋亡情况。结果：拯阳汤含药血清对阳虚小鼠腹腔巨噬细胞活化作用及其分泌的IL-1活性在一定剂量范围内均具有促进作用，但大剂量的促进作用有减弱趋势；对脾脏T淋巴细胞增殖活性及其分泌IL-2活性在一定剂量范围内亦均具有促进作用，大剂量的促进作用亦有减弱趋势；对胸腺细胞凋亡的抑制作用在一定剂量范围内成正比，剂量越大，作用越强。结论：拯阳汤含药血清对阳虚小鼠腹腔MФ的活化及其分泌的IL-1活性、脾脏T淋巴细胞增殖及其分泌的IL-2在一定剂量范围内具有促进作用，对阳虚小鼠的胸腺细胞凋亡的抑制作用在一定剂量的范围内呈量一效关系。     

玉屏风散发酵液对小鼠腹腔巨噬细胞免疫功能影响的血清药理学研究

目的:采用血清药理学方法,观察玉屏风散煎剂及发酵液对小鼠腹腔巨噬细胞的免疫功能的影响.方法:3组小鼠按37.0g/kg体重分别用玉屏风散发酵液及水煎剂连续灌胃3d,空白对照组用自来水灌胃.于末次给药后lh眼球采血制备含药血清及空白对照血清,观察含药血清对小鼠腹腔巨噬细胞吞噬功能和NO生成量的影响.结果:玉屏风散发酵液与煎剂都可增强小鼠腹腔巨噬细胞的吞噬功能及NO的生成量,且玉屏风散发酵液的作用强于煎剂.结论:玉屏风散发酵液可明显增强小鼠的免疫功能.     

鹅不食草挥发油抗变态反应血清药理学研究

目的:用血清药理学方法研究鹅不食草挥发油对抗原诱导RBL-2H3细胞释放组胺、β-氨基已糖苷酶,对刀豆蛋白(Con-A)诱导小鼠脾细胞增殖、诱导脾淋巴细胞分泌IL-4的影响,探讨其抗变态反应的可能机制。方法:ELISA法测定鹅不食草挥发油含药血清对抗原诱导RBL-2H3释放组胺、β-氨基已糖苷酶的影响;MTT法测定鹅不食草挥发油含药血清对Con-A诱导脾细胞增殖的影响,ELISA法测定鹅不食草挥发油含药血清对Con-A诱导脾淋巴细胞分泌IL-4的影响。结果:与空白血清相比,鹅不食草挥发油0.5～2h含药血清(9g/kg生药量,10%含药血清)均具有抑制抗原诱导RBL-2H3释放组胺、β-氨基已糖苷酶,抑制Con-A诱导小鼠脾细胞增殖、分泌IL-4的作用。结论:鹅不食草挥发油抗变态反应作用机制可能与抑制炎症介质释放,抑制细胞免疫功能有关。     

四倍体黄芩新品系D20与甘肃地产二倍体黄芩的抗炎作用比较

目的:四倍体黄芩新品系D20水提物与甘肃地产二倍体黄芩水提物的抗炎作用比较。方法:采用二甲苯致小鼠耳廓肿胀、小鼠棉球肉芽肿、小鼠腹腔毛细血管通透性、LPS刺激的巨噬细胞急性炎症及大鼠角叉菜胶致足肿胀实验,观察四倍体黄芩新品系D20水提物与甘肃地产二倍体黄芩水提物的抗炎作用。实验设空白组、地塞米松组(30mg/kg)、双黄连粉针剂组(45mg/kg)、二倍体黄芩低剂量(5g/kg)、二倍体黄芩高剂量组(10g/kg)、四倍体黄芩低剂量组(5g/kg)、四倍体黄芩高剂量组(10g/kg)。黄芩水提物等体积经口灌胃给药,地塞米松和双黄连粉针剂腹腔注射给药。结果:四倍体黄芩新品系D20水提物与甘肃地产二倍体黄芩水提物均可不同程度减轻二甲苯致小鼠耳廓肿胀度和小鼠棉球肉芽肿,降低小鼠腹腔毛细血管通透性和LPS刺激的巨噬细胞急性炎症,抑制大鼠角叉菜胶所致的足肿胀,二者的作用强度基本无差异。结论:四倍体黄芩新品系D20抗炎作用与甘肃地产二倍体黄芩水提物无差异。     

大黄牡丹汤含药血清对巨噬细胞释放炎症因子的影响

目的研究大黄牡丹汤临床治疗炎症性疾病的部分机制。方法常规制备大黄牡丹汤水煎剂,分大、中、小剂量灌胃干预小鼠并制备含药血清,在96孔板中将含药血清与小鼠腹腔巨噬细胞共孵,收集培养上清,采用Griess比色法检测巨噬细胞NO分泌量、ELISA法检测巨噬细胞TNF-α、IL-1β、IL-6分泌量。结果与正常血清对照组比较,大黄牡丹汤含药血清能剂量依赖性地促进正常巨噬细胞分泌NO、TNF-α、IL-1β和IL-6(P0.01);能抑制LPS活化的巨噬细胞分泌NO、TNF-α和IL-1β(P0.05或P0.01),但对LPS活化的巨噬细胞分泌IL-6无显著影响(P0.05)。结论大黄牡丹汤对活化前后的巨噬细胞分泌炎症因子的干预作用,可能是该方调控炎症反应的主要机制之一。     

傣药嘿盖贯不同提取物对小鼠耳廓肿胀抑制作用的研究

目的研究傣药嘿盖贯不同提取物(水提物、醇提物)对小鼠耳廓肿胀的抑制作用。方法通过二甲苯致小鼠耳廓肿胀急性炎症动物模型,评价傣药嘿盖贯提取物的抗炎效果。结果与模型对照组比较,嘿盖贯水提物高剂量组、水提物低剂量组、醇提物高剂量组的肿胀度具有极显著差异(P0.01),醇提物低剂量组差异显著(P0.05)。与阳性对照组比较,嘿盖贯水提物高剂量组、醇提物高剂量组的肿胀抑制率相近。结论说明傣药嘿盖贯水提物、醇提物具有明显的抑制炎症的作用。     

当归饮子含药血清对致敏小鼠脾淋巴细胞释放白三烯B4的影响研究

目的:观察当归饮子含药血清对致敏小鼠脾淋巴细胞释放白三烯B4(LTB4)的影响。方法:BALB/c小鼠(SPF级)80只,其中10只用于制备致敏小鼠脾淋巴细胞悬液。其余随机分为空白对照组、当归饮子高剂量组、当归饮子低剂量组、咪唑斯汀组、孟鲁司特组、氯雷他定组、地塞米松组。分离致敏后小鼠的脾淋巴细胞,置于细胞培养板中。分别加入各组含药血清作预处理,再加入致敏原进行刺激。以ELISA法检测上清液白三烯B4含量。进行各组间比较。结果:与空白对照组比较,当归饮子高剂量组、当归饮子低剂量组、咪唑斯汀组、地塞米松组均可显著抑制小鼠脾淋巴细胞释放LTB4(P<0.01);当归饮子高剂量组、当归饮子低剂量组、咪唑斯汀组的抑制作用明显低于地塞米松组(P<0.05);同时当归饮子高剂量组的抑制作用强于当归饮子低剂量组、咪唑斯汀组(P<0.05)。结论:当归饮子含药血清对致敏小鼠脾淋巴细胞释放白三烯B4具有抑制作用,其抑制作用表现出一定的剂量依赖性。     

荭草提取物抗炎镇痛作用实验研究

目的探讨荭草提取物的抗炎镇痛作用。方法采用二甲苯致小鼠耳肿胀、蛋清致大鼠足跖肿胀2种急性炎症模型以及大鼠棉球肉芽肿慢性炎症模型,研究荭草水提物和醇提物ig给药的抗炎作用。采用热板法和醋酸扭体法致痛,研究荭草水提物和醇提物ig给药的镇痛作用。结果与模型组相比,荭草水提物和醇提物高、低剂量(生药7.5、3.75 g/kg)组显著抑制二甲苯所致小鼠耳肿胀(P<0.01),明显抑制蛋清所致大鼠足跖肿胀(P<0.05、0.01),明显抑制大鼠棉球肉芽肿(P<0.01)。荭草水提物和醇提物高、低剂量组给药后明显延长热板所致小鼠的痛阈值,与模型组比较差异显著(P<0.01);荭草水提物和醇提物高、低剂量组明显减少醋酸所致小鼠扭体反应次数,与模型组比较差异显著(P<0.01)。荭草水提物高、低剂量组给药后与相同剂量醇提物相比,各项指标差异显著(P<0.05、0.01)。结论荭草提取物ig给药具有抗炎、镇痛作用,且水提物作用强于醇提物。     

玉屏风散对小鼠腹腔巨噬细胞活化作用的血清药理学研究

目的：应用中药血清药理学方法，观察玉屏风散对小鼠腹腔巨噬细胞活化功能的影响。方法：9组小鼠按0.1ml/10g体重用玉屏风散水煎液灌胃，剂量相当于48.0g饮片／kg体重，于给药后15min、30min、1h、1.5h、2h、2.5h、3h、4h分别在无菌条件下取血，分离血清进行腹腔巨噬细胞活化作用检测。另设蒸馏水灌胃后的空白血清作对照组。结果：30min-90min含药血清对小鼠腹腔巨噬细胞均具有活化作用（P＜0．01）。结论：玉屏风散煎剂对腹腔巨噬细胞具有不同程度的活化作用。     

Stimulation of lymphocyte proliferation and inhibition of nitric oxide production by aqueous Urtica dioica extract

The immunomodulatory and antiinflammatory activities of aqueous Urtica dioica extract were investigated for their effect on the mitogenic response of murine splenocytes and nitric oxide production by murine peritoneal macrophages in vitro. It was found that this extract stimulated the proliferation of T-lymphocytes and suppressed NO production in lipopolysaccharide-stimulated macrophages without affecting cell viability.     

Screening of Chinese herbal drug extracts for inhibitory activity on nitric oxide production and identification of an active compound of Zanthoxylum bungeanum

Sixty-eight water and methanol extracts from 34 Chinese herbal drugs, most of which are used for inflammatory diseases, were screened for their inhibitory effects on nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated J774.1 macrophages and in LPS/interferon (IFN)-gamma-stimulated mouse peritoneal exudate macrophages. Among the extracts, methanol extracts of Myristica fragrans, Plantago asiatica, Rubia cordifolia, and Zanthoxylum bungeanum showed significant inhibition in J774.1 macrophages, while in mouse peritoneal exudate macrophages, water extracts of Ru. cordifolia and Scutellaria baicalensis and methanol extracts of Angelica megaphylla, My. fragrans, and Z. bungeanum inhibited the NO production. Among them, inhibition of water extract of Sc. baicalensis was found to be mainly due to direct scavenging of NO radicals, through an examination of its scavenging activity on PAPA NONOate-generated NO radicals, while water extract of Ru. cordifolia and methanol extracts of An. megaphylla, My. fragrans, P. asiatica, and Z. bungeanum showed inhibition on iNOS mRNA expression. At last, an inhibitory compound on iNOS mRNA expression was isolated from a methanol extract of Z. bungeanum and identified as 4-O-beta-D-glucopyranosyldihydroferulic acid by NMR spectral analyses and chemical synthesis.     

碎米花杜鹃原花青素A-1对小鼠免疫细胞的影响

目的:研究碎米花杜鹃提取物乙酸乙酯部分分离得到的化合物原花青素A-1(Proanthocyanidin A-1,简称PAA-1)的免疫调节活性。方法:采用MTT法,通过PAA-1体外刺激,研究了其对小鼠脾淋巴细胞和腹腔巨噬细胞的增殖、NK细胞杀伤活性及对巨噬细胞吞噬活性和释放效应分子NO的影响。结果:PAA-1在浓度为5～100 mg/L时能刺激脾细胞增殖,增加腹腔巨噬细胞能量代谢能力,增强巨噬细胞对中性红的吞噬能力及NO的分泌量,激活自然杀伤细胞(NK细胞)对YAC-1细胞的杀伤能力。结论:PAA-1能增强小鼠的免疫功能。     

Acanthopanax senticosus inhibits nitric oxide production in murine macrophages in vitro and in vivo

Excess nitric oxide (NO) production has been implicated in inflammatory diseases. The present study investigated the inhibitory effect of the stem bark extract of Acanthopanax senticosus (A. senticosus) on NO production in murine macrophages in vitro and in vivo. In vitro exposure of RAW264.7 cells to 1, 10, 50, 100, 250, 500 and 1000 microg/mL of A. senticosus extract significantly suppressed NO production induced by lipopolysaccharide (LPS) and interferon gamma (IFN-gamma) in a dose-dependent manner. In vitro exposure of mouse resident peritoneal macrophages to 1, 10, 100 and 1000 microg/mL of A. senticosus extract significantly suppressed NO production induced by LPS and IFN-gamma in a dose-dependent manner. In vivo administration of A. senticosus extract (50, 100 and 200 mg/kg) to KM mice dose-dependently inhibited LPS and IFN-gamma induced production of NO in isolated mouse peritoneal macrophages ex vivo. Exposure to A. senticosus extract had no effect on cell viability and systemic toxicity. The results demonstrated that the stem bark extract of A. senticosus extract inhibits NO production in murine macrophages in vitro and in vivo.     

罗红霉素对迟发型变态反应的抑制作用

目的：探讨罗红霉素对迟发型变态反应及其相关细胞的影响。方法：整体实验：以绵羊红细胞（SRBC）诱导小鼠迟发型变态反应（DTH）。离体实验：分离培养小鼠脾细胞和大鼠腹腔巨噬细胞，采用M3T比色法测定细胞数；以Gness法测定细胞释放的NO水平。结果：罗红霉素10和20mg／kg于效应相经口给药显著地抑制了SRBC诱导的迟发型变态反应，并使因SRBC免疫增高的淋巴细胞体外存活率恢复至正常水平；罗红霉素在体外能浓度依赖性地抑制ConA所致的脾淋巴细胞增殖及腹腔巨噬细胞释放NO；体内给药显著地抑制了脾淋巴细胞对ConA的增殖反应能力。结论：罗红霉素对SRBC-DTH有显著的抑制作用，其抑制效应可能与降低脾淋巴细胞的存活、增殖及巨噬细胞释放NO等有关。     

Production of nitric oxide in mouse peritoneal macrophages after priming with interferon-gamma by the stem of Sinomenium acutum.

The present study demonstrates that the aqueous extract of Sinomenium acutum stem (SSAE) produces nitric oxide (NO) upon treatment with recombinant interferon gamma (rIFN-gamma) in mouse peritoneal macrophages. Apparently SSAE has no effect on NO production by itself. This production is dependent on L-arginine and can be inhibited by the L-arginine analogue N(G)-monomethyl-L-arginine. The increased production of NO from rIFN-gamma plus SSAE-stimulated cells was decreased by the treatment of protein kinase C inhibitor. Tumor necrosis factor-alpha (TNF-alpha) has been shown to stimulate the oxidative metabolism of L-arginine to produce NO. Mouse peritoneal macrophages secrete high levels of TNF-alpha after incubation with rIFN-gamma plus SSAE. In addition, SSAE-induced NO production is progressively inhibited by anti-murine TNF-alpha neutralizing antibody. These results show that the capacity of SSAE to increase NO production from rIFN-gamma-primed mouse peritoneal macrophages is the result of SSAE-induced TNF-alpha secretion.     

亮菌多糖IPS-B2对小鼠腹腔巨噬细胞活性及其相关基因转录的影响

目的：观察亮菌多糖IPS-B2对巨噬细胞分泌细胞因子、一氧化氮(NO)生成以及巨噬细胞中白介素-1β(IL-1β)，白介素-6(IL-6)，肿瘤坏死因子-α(TNF-α)和NO合成关键酶诱导型一氧化氮合酶(iNOS)基因转录的影响。方法：采用ELISA和Griess法检测IPS-B2对小鼠腹腔巨噬细胞生成细胞因子IL-1β，IL-6，TNF-α和细胞毒效应分子NO的影响；采用SYBR Green Ⅰ指示的Real-time RT-PCR技术研究IPS-B2对小鼠腹腔巨噬细胞中IL-1β，IL-6，TNF-α和NO合成关键酶iNOS基因在mRNA转录水平上的作用。结果：IPS-B2能显著提高小鼠腹腔巨噬细胞培养上清液中IL-1β，IL-6，TNF-α和NO的含量，增强IL-1β，IL-6，TNF-α和NO合成关键酶iNOS基因的转录水平。结论：IPS-B2可能是通过提高巨噬细胞分泌生物活性物质，并促进此类生物活性物质的基因转录，从而实现其抗肿瘤的重要作用。     

Absolute stereostructures of new arborinane-type triterpenoids and inhibitors of nitric oxide production from Rubia yunnanensis

The aqueous acetone extract from the roots of a Chinese herbal medicine, Rubia yunnanensis, showed a potent inhibitory effect on nitric oxide production in lipopolysaccharide-activated macrophages. Five new arborinane-type triterpenes, rubianols-a (1), -b (2), -c (3), -d (4), and -e (5), and a new arborinane-type triterpene glycoside, rubianoside I (6), were isolated from the herbal crude extract together with 10 known compounds. The absolute stereostructures of 1-6 were determined on the basis of chemical and physicochemical evidence, including the application of the modified Mosher's method. The effects of the isolated constituents on nitric oxide production in lipopolysaccharide-activated macrophages were examined, and several triterpenes were found to show inhibitory activity.     

当归、丹参和川芎嗪注射液对腹膜透析腹腔巨噬细胞功能的干预作用

目的：研究当归、丹参和川芎嗪注射液（简称中药）对腹膜透析（PD）腹腔巨噬细胞（MC）功能的影响。方法：（1）将MC置于含当归、丹参和川芎嗪的腹膜透析液（CDS）的培养液中培养24h，测定3个中药组、CDS组和对照组MC的一氧化氮（NO）含量和对四甲基偶氮唑盐（MTT）还原能力。（2）MC分别置于含2、10、100μg/ml的3个中药的CDS中培养24h，观察不同浓度中药对MC吞噬能力、NO含量和MTT还原能力的影响，对照组为等量培养液代替CDS。结果：CDS组MC的NO含量和MTT还原能力均明显低于对照组（P＜0．01）；与CDS组比较，川芎嗪组的MC NO含量和3个中药组MC MTT还原能力均有明显提高（P＜0．01），并随着培养液中的中药浓度的提高，3个中药组MC吞噬能力和NO含量均有显著增加。结论：在CDS中加中药能改善腹膜腔MC的防御功能，降低腹膜炎的发生率，对提高PD疗效有重要意义。     

Acanthopanax senticosus suppresses reactive oxygen species production by mouse peritoneal macrophages in vitro and in vivo

Excess production of reactive oxygen species by macrophages has been implicated in many inflammatory diseases. The present study investigated the inhibitory effect of the stem bark extract of Acanthopanax senticosus on the production of superoxide anion and hydrogen peroxide in mouse peritoneal macrophages in vitro and in vivo. Exposure of mouse peritoneal macrophages to A. senticosus extract significantly suppressed superoxide anion production induced by zymosan in a dose-dependent manner. Similarly, exposure of mouse peritoneal macrophages to A. senticosus extract significantly inhibited hydrogen peroxide production induced by phorbol 12-myristate 13-acetate (PMA) in a dose-dependent manner. Intraperitoneal administration of A. senticosus extract to KM mice reduced the ex vivo production of zymosan induced-superoxide anion and PMA-induced hydrogen peroxide by their peritoneal macrophages. Exposure to A. senticosus extract did not affect the cell viability or systemic toxicity. A. senticosus inhibited reactive oxygen species production by mouse peritoneal macrophages in vitro and in vivo and may be partly responsible for the antiinflammatory function.