清开灵注射液诱发类过敏反应作用及其机制研究

目的 研究清开灵注射液诱发类过敏反应的作用及其机制。方法 采用BALB/c小鼠和BN大鼠,观察给予清开灵注射液后小鼠皮肤血管渗透性以及大鼠平均动脉血压等类过敏指标的变化;利用RBL-2H3大鼠嗜碱性粒细胞白血病细胞,观察清开灵注射液作用后磷脂酰肌醇3-激酶（PI3K）活性以及Rac1、PAK1、LIMK1和人肌动蛋白素（Cofilin）蛋白含量的变化。结果 5 mL·kg^-1清开灵注射液即可引起小鼠以血管通透性增高为主要表现的较为轻微的类过敏反应,而7.5 mL·kg^-1和15 mL·kg^-1清开灵注射液均可引起较为严重的类过敏反应,其反应呈剂量依赖性;另外,5,7.5和15 mL·kg^-1清开灵注射液可在一定时间范围内剂量依赖性地引起大鼠平均动脉血压降低。清开灵注射液体外可诱导RBL-2H3细胞发生类过敏反应,PI3K激酶活性增加,Rac1、PAK1、LIMK1和Cofilin蛋白含量增高,且具有剂量依赖性。结论 小鼠和大鼠体内类过敏反应检查法和RBL-2H3细胞中PI3K激酶及Rac1、PAK1、LIMK1、Cofilin蛋白含量检查法可作为类过敏反应检查方法。初步认定清开灵注射液诱导类过敏反应的发生可能是基于PI3K-Rac1细胞运动信号转导通路。     

桑寄生抗炎作用的初步实验研究

目的 研究桑寄生水提物（简称SJS）对急性、亚急性及慢性炎症的影响。方法 采用冰醋酸、二甲苯复制致小鼠急性炎症模型,羧甲基纤维素（CMC）囊复制小鼠亚急性炎症模型和棉球皮下埋植复制小鼠慢性炎症模型。将动物随机、均衡分成空白对照组、模型对照组、阳性对照组、桑寄生水提物高和低剂量组,通过测定腹腔冲洗液光密度（OD）值和小鼠耳廓肿胀度来评价桑寄生水提物对急性炎症期渗出和毛细血管通透性的影响;计数羧甲基纤维素囊中白细胞数量来评价桑寄生水提物对亚急性炎症期白细胞游走的影响;测定棉球肉芽组织指数来评价桑寄生水提物对慢性炎症期肉芽组织增生的影响,另取脾脏和胸腺称重,观察连续给药后桑寄生水提物对免疫器官重量的影响。结果 与模型对照组比较,桑寄生水提物高、低剂量均能显著降低冰醋酸所致小鼠腹腔毛细血管通透性增加(P<0.05～0.01),高剂量能显著减轻二甲苯致小鼠耳廓肿胀率（P<0.05）;高剂量能显著减少羧甲基纤维素诱导白细胞游出数量（P<0.001）;高剂量能显著降低棉球肉芽肿指数（P<0.01）;桑寄生水提物高、低剂量对脾脏和胸腺重量均无显著影响（P>0.05）。结论 桑寄生水提物对炎症不同时期均有明显的抑制作用,且连续给药对免疫器官的重量无影响。     

槐白皮水提物对免疫抑制小鼠的免疫作用

目的 探讨槐白皮水提物对免疫抑制小鼠免疫功能的影响。方法 昆明小鼠随机分为正常组,模型组,香菇多糖组和槐白皮水提物低、中、高剂量组,除正常组外,其余各组连续3 d腹腔注射环磷酰胺复制免疫抑制模型。造模后连续灌胃给药14 d,血细胞计数仪测定小鼠外周血白细胞数目、称量并计算免疫器官系数、MTT法测定脾淋巴细胞增殖和NK细胞活性,ELISA检测血清IL-2、IFN-γ和TNF-α水平。结果 槐白皮水提物中、高剂量组明显提高免疫抑制小鼠的外周血白细胞水平和免疫器官系数,显著增加LPS诱导的脾淋巴细胞增殖,显著增强脾NK细胞活性,提高免疫抑制小鼠血清IL-2、IFN-γ水平;槐白皮水提物高剂量组明显提高ConA诱导的脾淋巴细胞增殖,显著提高免疫抑制小鼠血清TNF-α水平。结论 槐白皮水提物可以提高免疫抑制小鼠的免疫功能。     

基于实时细胞分析技术评价注射用益气复脉(冻干)类过敏反应

目的 建立中药注射剂体外类过敏反应评价方法,快速评价不同批次注射用益气复脉（冻干）类过敏反应表现。方法 体外培养嗜碱性白血病细胞株RBL-2H3细胞,选择Compound 48/80为阳性药,采用实时细胞分析（real-time cell analysis,RTCA）系统检测药物干预后引起的细胞指数（CI）值变化,并利用甲苯胺蓝、鬼笔环肽染色观察细胞形态、骨架变化,以及检测组胺和β-已糖苷酶释放量验证RBL-2H3细胞的脱颗粒情况。选择20个批次的注射用益气复脉（冻干,100 μg/mL）作用于RBL-2H3细胞,进行基于RTCA技术的类过敏反应评价。结果 阳性药Compound 48/80（20 μg/mL）能够使RBL-2H3细胞的CI值在加药后30 min内呈先快速上升后下降趋势;形态学研究发现,Compound 48/80使细胞形态和细胞骨架均发生明显改变,发生明显的脱颗粒现象;组胺和β-己糖苷酶释放实验进一步证实Compound 48/80导致炎症介质的释放,引起了明显的脱颗粒现象;提示RTCA系统可以用于快速敏感的评价RBL-2H3细胞脱颗粒。不同批次的注射用益气复脉（冻干）对RBL-2H3细胞CI值无明显影响,提示所选批次为合格批次,无类过敏反应现象的发生。结论 建立了一套基于RTCA系统的类过敏反应体外快速评价技术,可用于注射用益气复脉（冻干）等中药注射剂类过敏反应的体外快速评价。     

聚山梨酯80对多源肥大细胞脱颗粒的影响研究

目的 通过比较不同浓度聚山梨酯80溶液对RBL-2H3、P815、Ku812细胞脱颗粒的影响,探究聚山梨酯80的量与过敏反应的关系,进而筛选出一套灵敏、稳定的体外肥大细胞脱颗粒模型。方法 体外培养RBL-2H3、P815和Ku812细胞,测定3株细胞的生长曲线,在细胞生长对数期,以不同浓度（0.04、0.20、1.00、5.00、10.00、20.00、40.00、80.00 mg/mL）聚山梨酯80分别刺激3株细胞,采用中性红染色法显微镜下观察不同浓度聚山梨酯80溶液对3株细胞脱颗粒的影响,并计算脱颗粒百分率,同时以化学发光法分别检测组胺和β-氨基己糖苷酶释放率,以酶联免疫吸附法测定类胰蛋白酶的释放量;并进一步检测聚山梨酯80对人的肥大细胞IgE释放的影响。结果 3株肥大细胞脱颗粒模型中,各细胞的脱颗粒指标均随聚山梨酯80浓度的升高而升高。相同浓度聚山梨酯80对人源、大鼠、小鼠肥大细胞的类胰蛋白酶释放量无较大统计学差异;与RBL-2H3细胞系比较,P815细胞和组胺释放率显著降低（P<0.05、0.01）,Ku812细胞组胺释放率和β-氨基己糖苷酶释放率显著升高（P<0.05、0.01）,Ku812细胞脱颗粒最灵敏。但在实验过程中发现Ku812细胞模型稳定性、可重复性较差,而RBL-2H3细胞稳定性、可重复性均优于Ku812和P815细胞。0.04~80.00 mg/mL的聚山梨酯80溶液作用于Ku812细胞产生的IgE均低于检测限。结论 聚山梨酯80诱导的过敏反应可能是不经过IgE介导的类过敏反应,随着聚山梨酯80浓度的增大,3个细胞模型脱颗粒现象显著,相比于Ku812和P815细胞,RBL-2H3细胞更适合作为体外肥大细胞脱颗粒检测模型。     

牛樟芝胶囊对大鼠四氯化碳肝损伤的保护作用研究

摘 要 目的: 观察牛樟芝胶囊对四氯化碳（CCl-4）所致大鼠肝损伤的保护作用,并与甘草酸二铵胶囊作用效果进行比较。方法: 用CCl-4制备大鼠中毒性肝损伤模型,观察牛樟芝胶囊对大鼠血清中ALT、AST和乳酸脱氢酶（LDH）变化的影响,以及对肝脏病理损伤的作用。结果:模型对照组与阴性对照组比较,动物体质量降低,肝脏绝对质量（15.13±2.01）g和相对质量（4.32±0.36）g/100 g增高,ALT（189.9±50.4 )IU·L^-1、AST（249.1±47.6)I U·L^-1、LDH（1 740.8±522.0)IU·L^-1均显著增高（P<0.05）,肝组织病理损伤明显加重,肝损伤模型成立。中剂量牛樟芝胶囊（1.08 g·kg^-1）可使肝损伤大鼠的ALT（126.7±31.4）IU·L^-1、AST（175.2±38.9）IU·L^-1、LDH（1 013.7±342.5）IU·L^-1明显降低（P<0.05）;高剂量牛樟芝胶囊（1.62 g·kg^-1）可使肝损伤大鼠的ALT（138.0±50.0）IU·L^-1和AST（189.1±56.7）IU·L^-1明显降低（P<0.05）,肝组织病理损伤明显改善（P<0.05）,优于甘草酸二铵胶囊。 结论:牛樟芝对大鼠CCl-4肝损伤有保护作用。     

连钱草提取物对炎症递质的影响

目的 研究连钱草提取物对炎症递质（PGE2、组胺、5 羟色胺等）的影响，探讨连钱草提取物的抗炎作用机制。方法分别制作二甲苯所致小鼠耳肿胀、醋酸致小鼠腹腔毛细血管通透性增加、角叉菜胶致小鼠足肿胀、蛋清致小鼠足组织炎症等模型，考察连钱草水提物（1 g·mL^-1）和醇提物（1 g·mL^-1）对模型小鼠组织中炎症递质的影响，测定炎症组织中PGE2、5-羟色胺和组胺相对含量。结果连钱草水提物与醇提物对二甲苯致小鼠耳肿胀均有显著的抑制效果；对醋酸所致小鼠腹腔毛细血管通透性亦有显著抑制效果；连钱草醇提取物对角叉菜胶所致小鼠肿胀足组织中PGE2释放无抑制作用，而水提物则有明显的抑制作用；连钱草水提物对蛋清所致小鼠肿胀足组织中组胺的释放有明显抑制作用，但连钱草醇提物无该作用；连钱草水提物对蛋清所致小鼠肿胀足组织中5 羟色胺的释放有明显抑制作用（P＜0.05），连钱草醇提物作用较水提物小。结论 连钱草提取物通过抑制内源性炎症递质5 羟色胺和组胺的释放而发挥抗炎作用，水提物抗炎作用较醇提物明显。     

还阳参醇提物止咳祛痰平喘及抗炎作用的实验研究

目的 探讨还阳参醇提物对实验动物的止咳、祛痰、平喘及抗炎作用。方法 分别采用氨水引咳法、气管酚红排泄法、组织胺致喘息法观察还阳参醇提物的止咳、祛痰、平喘作用;采用二甲苯致小鼠耳肿胀和蛋清致大鼠足趾肿胀法观察还阳参醇提物抗急性炎症的作用;气管内注入脂多糖法建立大鼠慢性支气管炎模型,观察受试物对模型大鼠肺组织的病理学改变。结果 还阳参醇提物能显著减少小鼠的咳嗽次数、增加小鼠气道酚红排泄量、延长组织胺所致豚鼠喘息的潜伏期、可减轻蛋清所致的大鼠足趾肿胀度,与对照组比较差异具有显著性（P＜0.05或0.01）;还阳参醇提物能减轻慢性支气管炎大鼠黏膜上皮细胞的坏死和脱落,抑制黏膜层和黏膜下层炎性细胞浸润。结论 还阳参醇提物有明确的止咳、祛痰、平喘及抗炎作用,对慢性支气管炎大鼠模型的支气管黏膜有保护作用。     

冬虫夏草菌丝体水提醇沉物体外抗肿瘤活性研究

目的 研究冬虫夏草菌丝体水提醇沉物的体外抗肿瘤活性及其机制。方法 采用热水浸提-醇沉法得到水提物,高效液相色谱仪测定水提醇沉物中腺苷的量;采用MTT法测定其抗肿瘤活性,利用流式细胞仪结合碘化丙锭染色法检测其对细胞周期的抑制。结果 实验表明水提醇沉物能抑制人肝癌HepG2细胞株及大细胞肺癌NCI-H460细胞株的增殖,并呈浓度相关性;半数抑制浓度（IC₅₀）值分别为（1.49±0.19）和（1.67±0.27）mg/mL。细胞周期分析表明,水提醇沉物分别阻滞HepG2及NCI-H460细胞周期于G₂/M期、S期,并可诱导上述两种细胞发生凋亡。结论 冬虫夏草水提醇沉物通过阻滞HepG2及NCI-H460细胞周期循环,诱导其凋亡,从而表现出良好的增殖抑制活性。为深入研究冬虫夏草菌丝体水提醇沉物抗肿瘤的机制提供了实验证据。     

芪防鼻敏颗粒抗炎、抗过敏作用研究

摘 要 目的：初步探讨中药复方芪防鼻敏颗粒的抗炎、抗过敏作用。方法: 采用二甲苯致小鼠耳廓肿胀法和大鼠棉球肉芽组织形成法观察芪防鼻敏颗粒的抗炎作用,采用大鼠同种被动皮肤过敏反应实验和小鼠耳异种被动皮肤过敏反应观察芪防鼻敏感颗粒的抗过敏作用。结果: 与空白组比较,芪防鼻敏颗粒各剂量组可显著降低二甲苯所致小鼠耳廓肿胀度（P<0.01或0.05）,对大鼠棉球肉芽组织增生无明显抑制作用。与空白组比较,芪防鼻敏颗粒各剂量组可明显降低卵蛋白所致大鼠皮肤蓝斑A值（P<0.01或0.05）,芪防鼻敏颗粒高剂量组可显著降低小鼠耳廓蓝染程度（P<0.05）。结论:芪防鼻敏颗粒具有一定抗炎、抗过敏作用。     

Inhibitory mechanism of saponins derived from roots of Platycodon grandiflorum on anaphylactic reaction and IgE-mediated allergic response in mast cells

The purpose of this study was to investigate the protective effects of saponins isolated from the root of Platycodi Radix (Changkil saponins: CKS) anti-allergic effects in mice and mast cells. Oral administration of CKS inhibited the dinitrophenyl (DNP)–IgE antibody-induced systemic PCA reaction in mice. CKS reduced the β-hexosaminidase and histamine release from anti-DNP–IgE-sensitized RBL–2H3 cells. In addition, CKS inhibited the IgE antibody-induced increases in IL-4 and TNF-α production and expression in RBL-2H3 cells. In order to explore the inhibitory mechanism of CKS in PCA and mast cell degranulation, we examined the activation of intracellular signaling molecules. CKS suppressed DNP–IgE antibody-induced Syk phosphorylation. Further downstream, CKS also inhibited the phosphorylation of Akt and MAP kinases. Taken together, the in vivo/in vitro anti-allergic effects of CKS suggest possible therapeutic applications for this agent in allergic diseases through the inhibition of inflammatory cytokines and Syk-dependent signaling cascades.     

Houttuynia cordata water extract suppresses anaphylactic reaction and IgE-mediated allergic response by inhibiting multiple steps of FcεRI signaling in mast cells

Houttuynia cordata has been used as a traditional medicine in Korea and is known to have antioxidant, anti-cancer and anti-allergic activities. The precise effect of H. cordata, however, remains unknown. In this study, we investigated the effects of H. cordata water extract (HCWE) on passive cutaneous anaphylaxis (PCA) in mice and on IgE-mediated allergic response in rat mast RBL-2H3 cells. Oral administration of HCWE inhibited IgE-mediated systemic PCA in mice. HCWE also reduced antigen (DNP-BSA)-induced release of β-hexosaminidase, histamine, and reactive oxygen species in IgE-sensitized RBL-2H3 cells. In addition, HCWE inhibited antigen-induced IL-4 and TNF-α production and expression in IgE-sensitized RBL-2H3 cells. HCWE inhibited antigen-induced activation of NF-κB and degradation of IκB-α. To investigate the inhibitory mechanism of HCWE on degranulation and cytokine production, we examined the activation of intracellular FcεRI signaling molecules. HCWE suppressed antigen-induced phosphorylation of Syk, Lyn, LAT, Gab2, and PLC γ2. Further downstream, antigen-induced phosphorylation of Akt and MAP kinases (ERK1/2 and JNK1/2 but not p38 MAP kinase) were inhibited by HCWE. Taken together, the in vivo/in vitro anti-allergic effect of HCWE suggests possible therapeutic applications of this agent in inflammatory allergic diseases through inhibition of cytokines and multiple events of FcεRI-dependent signaling cascades in mast cells.     

Ethyl acetate extract of Psidium guajava inhibits IgE-mediated allergic responses by blocking FcεRI signaling

Psidium guajava (P. guajava) is an important food crop and medicinal plant with antioxidant, anti-inflammatory, and anti-allergic activities, supporting its traditional uses. However, its precise effects remain unknown. We investigated the effects of P. guajava ethyl acetate extract (PGEA) on IgE-mediated allergic responses in rat mast RBL-2H3 cells. PGEA reduced antigen (DNP-BSA)-induced release of β-hexosaminidase and histamine in IgE-sensitized RBL-2H3 cells. In addition, it inhibited antigen-induced IL-4 and TNF-α mRNA expression and protein production in IgE-sensitized RBL-2H3 cells. PGEA also suppressed antigen-induced COX-2 mRNA and protein expression in these cells, as well as antigen-induced activation of NFAT and reactive oxygen species. Moreover, it inhibited antigen-induced activation of NF-κB and degradation of IκB-α. To identify the mechanisms underpinning the inhibition of degranulation and cytokine production by PGEA, we examined the activation of intracellular FcεRI signaling molecules. PGEA suppressed antigen-induced phosphorylation of Syk, LAT, Gab2, and PLCγ2 but not Lyn, and inhibited antigen-induced phosphorylation of downstream signaling intermediates including MAP kinases and Akt. Collectively, the anti-allergic effects of PGEA in vitro suggest its possible therapeutic application to inflammatory allergic diseases, in which its inhibition of inflammatory cytokine production and FcεRI-dependent signaling events in mast cells may be hugely beneficial.     

Tofacitinib suppresses mast cell degranulation and attenuates experimental allergic conjunctivitis

BackgroundAllergic conjunctivitis (AC), a common eye inflammation that affects patients’ health and quality of life, is still a therapeutic challenge for ophthalmologists. Tofacitinib, a new Janus kinase (JAK) inhibitor, has been successfully used in the treatment of several disorders. Nonetheless, its effect in AC and the potential anti-allergic mechanisms are still unclear. The objective of the current study was to explore the roles of tofacitinib in preventing AC and elucidate the potential underlying mechanisms.MethodsTofacitinib was used topically in BALB/c mice with experimental allergic conjunctivitis (EAC). Ocular allergic symptoms and biological modifications were examined. To assess the anti-allergic mechanisms of tofacitinib, RBL-2H3 cells and HUVECs were cultured in vitro. The inhibitory effects and mechanisms of tofacitinib were studied and measured by real-time quantitative PCR, ELISA, western blot analysis, and flow cytometry.ResultsTopical administration of tofacitinib reduced the clinical symptoms of OVA-induced EAC, with a substantial mitigation in inflammatory cell infiltration, histamine release, and TNF-α mRNA as well as IL-4 mRNA expression. In vitro, tofacitinib repressed the degranulation and cytokine production in RBL-2H3 cells and reduced histamine-induced vascular hyperpermeability. The underlying mechanism might involve the downregulation of phosphorylation of JAK3/STATs signaling molecules in RBL-2H3 cells and HUVECs.ConclusionsOur findings provide evidence that tofacitinib prevented EAC by targeting the JAK3/STATs pathway. We recommend the use of tofacitinib as an innovative approach for the treatment of AC.     

Emodin, a naturally occurring anthraquinone derivative, suppresses IgE-mediated anaphylactic reaction and mast cell activation

The high-affinity receptor for IgE (Fc?RI)-mediated activation of mast cells plays an important role in allergic diseases such as asthma, allergic rhinitis and atopic dermatitis. Emodin, a naturally occurring anthraquinone derivative in oriental herbal medicines, has several beneficial pharmacologic effects, such as anti-cancer and anti-diabetic activities. However, the anti-allergic effect of emodin has not yet been investigated. To assess the anti-allergic activity of emodin, in vivo passive anaphylaxis animal model and in vitro mouse bone marrow-derived mast cells were used to investigate the mechanism of its action on mast cells. Our results showed that emodin inhibited degranulation, generation of eicosanoids (prostaglandin D₂ and leukotriene C₄), and secretion of cytokines (TNF-α and IL-6) in a dose-dependent manner in IgE/Ag-stimulated mast cells. Biochemical analysis of the Fc?RI-mediated signaling pathways demonstrated that emodin inhibited the phosphorylation of Syk and multiple downstream signaling processes including mobilization of intracellular Ca²⁺ and activation of the mitogen-activated protein kinase, phosphatidylinositol 3-kinase, and NF-κB pathways. When administered orally, emodin attenuated the mast cell-dependent passive anaphylactic reaction in IgE-sensitized mice. Thus, emodin inhibits mast cell activation and thereby the anaphylactic reaction through suppression of the receptor-proximal Syk-dependent signaling pathways. Therefore, emodin might provide a basis for development of a novel anti-allergic drug.     

Anti-allergic actions of a Chinese patent medicine,huoxiangzhengqi oral liquid,in RBL-2H3 cells and in mice

ContextHuoxiangzhengqi oral liquid (HXZQ-OL), a traditional Chinese medicine formula, has antibacterial, anti-inflammation and gastrointestinal motility regulation effects.ObjectiveThe study investigates the anti-allergic activity and underlying mechanism of HXZQ-OL.Materials and methodsIgE/Ag-mediated RBL-2H3 cells were used to evaluate the anti-allergic activity of HXZQ-OL (43.97, 439.7 and 4397 μg/mL) in vitro. The release of cytokines and eicosanoids were quantified using ELISA. RT-qPCR was used to measure the gene expression of cytokines. The level of intracellular Ca²⁺ was measured with Fluo 3/AM. Immunoblotting analysis was performed to investigate the mechanism of HXZQ-OL. In the passive cutaneous anaphylaxis (PCA), BALB/c mice (5 mice/group) were orally administrated with HXZQ-OL (263.8, 527.6 and 1055 mg/kg/d) or dexamethasone (5 mg/kg/d, positive control) for seven consecutive days.ResultsHXZQ-OL not only inhibited degranulation of mast cells (IC₅₀, 123 μg/mL), but also inhibited the generation and secretion of IL-4 (IC₅₀, 171.4 μg/mL), TNF-α (IC₅₀, 88.4 μg/mL), LTC4 (IC₅₀, 52.9 μg/mL) and PGD2 (IC₅₀, 195.8 μg/mL). Moreover, HXZQ-OL suppressed the expression of IL-4 and TNF-α mRNA, as well as the phosphorylation of Fyn, Lyn and multiple downstream signalling proteins including MAPK and PI3K/NF-κB pathways. In addition, HXZQ-OL (527.5 mg/kg) attenuated the IgE-mediated PCA with 55% suppression of Evans blue exudation in mice.ConclusionsHXZQ-OL attenuated the activation of mast cell and PCA. Therefore, HXZQ-OL might be used as an alternative treatment for allergic diseases.     

Olopatadine: a drug for allergic conjunctivitis targeting the mast cell

Importance of the field: Ocular allergic diseases are characterized by specific activation of conjunctival mast cells with subsequent release of preformed and newly formed mediators. Mast cells are thus the first therapeutic target of ocular anti-allergic treatments.

Areas covered in this review: In this review, a Medline literature search was conducted on conjunctival mast cells, their role in ocular allergy and mast cell stabilization by ocular anti-allergic compounds.

What the reader will gain: Olopatadine hydrochloride, a mast cell stabilizer and histamine receptor antagonist, has been shown to inhibit mast cell activation in an in vitro model of human mast cell culture, reducing histamine and TNF-α release and upregulating proinflammatory mediators in conjunctival epithelial cells. In the in vivo conjunctival allergen challenge (CAC) model in allergic subjects, combined with objective evaluations of tear mediators and cytology, olopatadine reduced histamine tear levels and all aspects of allergic inflammation, confirming the positive clinical effects observed in active allergic patients.

Take home message: Mast cells play a central role in the pathogenesis of ocular allergy. The CAC model is ideal for assessing the mast cell stabilizing effects of anti-allergic compounds. This review of clinical studies demonstrates the superiority of olopatadine compared with other topical allergic drugs.     

Characteristics of Choline Transport Across the Blood-Brain Barrier in Mice: Correlation with In Vitro Data

Purpose. We examined the functional properties of choline transport across the blood-brain barrier (BBB) in mice. We compared the kinetic parameters and transport properties with those found in our in vitro uptake experiments using mouse brain capillary endothelial cells (MBEC4). Methods. The permeability coefficient-surface area product (PS) values of [³H]choline at the BBB were estimated by means of anin situ brain perfusion technique in mice.Results. [³H]Choline uptake was well described by a two-component model: a saturable component and a nonsaturable linear component. The [³H]choline uptake was independent of pH and Na⁺, but was significantly decreased by the replacement of Na⁺ with K⁺. Various basic drugs, including substrates and inhibitors of the organic cation transporter, significantly inhibited the [³H]choline uptake. These in situ (in vivo) results corresponded well to the in vitro results and suggest that the choline transporter at the BBB is a member of the organic cation transporter (OCT) family. Conclusion. The choline transport mechanism at the BBB is retained in MBEC4.     

Anti-allergic activity of sesquiterpenes from the rhizomes of <Emphasis Type="Italic">Cyperus rotundus</Emphasis>

From the 70% ethanol extract of the rhizomes of Cyperus rotundus (CRE), several major constituents including the sesquiterpene derivatives (valencene, nootkatone, and caryophyllene α-oxide), monoterpenes (β-pinene, 1,8-cineole, and limonene) and 4-cymene were isolated and examined for their anti-allergic activity in vitro and in vivo. In rat basophilic leukemia (RBL)-1 cells, the sesquiterpenes strongly inhibited 5-lipoxygenase-catalyzed leukotrienes production. In addition, they inhibited β-hexosaminidase release by antigen-stimulated RBL-2H3 cells, with valencene having the highest inhibitory effect. CRE inhibited leukotrienes production and β-hexosaminidase release at 300 μg/mL. It was also found that the most active sesquiterpene (valencene) and CRE inhibited β-hexosaminidase degranulation by inhibiting the initial activation reaction, Lyn phosphorylation, in IgE-stimulated RBL-2H3 cells. Moreover, CRE, valencene and nootkatone significantly inhibited the delayed-type hypersensitivity reaction in mice when administered orally at 50–300 mg/kg. In conclusion, C. rotundus and its constituents, valencene, nootkatone, and caryophyllene α-oxide, exert anti-allergic activity in vitro and in vivo. These sesquiterpenes, but not monoterpenes, certainly contribute to the anti-allergic activity of the rhizomes of C. rotundus.     

甘草配伍大戟的体外肝毒性研究

目的 研究甘草和大戟配伍的体外肝毒性。方法 采用显微观察法和MTT法检测不同浓度的甘草单煎液、大戟单煎液、甘草-大戟合煎液和甘草-大戟单煎混合液对人肝癌细胞HepG2增殖的影响,并比较大戟单煎液、甘草-大戟合煎液和甘草-大戟单煎混合液相当浓度下细胞毒性的大小。结果 大戟单用及大戟与甘草配伍均有细胞毒性,且呈剂量相关性;与大戟单煎液相比,甘草-大戟单煎混合液细胞毒性无明显差异,甘草-大戟合煎液细胞毒性减小。结论 甘草和大戟配伍导致大戟的体外肝毒性减小。