Suppression of lipopolysaccharide-induced inflammatory responses in RAW 264.7 murine macrophages by aqueous extract of Clinopodium vulgare L. (Lamiaceae)

Ethnopharmacological relevance

The wild basil Clinopodium vulgare L. is commonly used in Bulgarian folk medicine for treatment of irritated skin, mastitis- and prostatitis-related swelling, as well as for some disorders accompanied with significant degree of inflammation (e.g. gastric ulcers, diabetes, and cancer).

Aim of study

To determine the effect of aqueous extract of Clinopodium vulgare L. on LPS-induced inflammatory responses of murine RAW 264.7 macrophages.

Materials and methods

Cell cytotoxicity was evaluated by MTT assay. Protein expression levels were monitored by Western blot analysis. Production of NO and PGE₂ was measured by the Griess colorimetric method and enzyme immunoassay, respectively. Activation of MMP-9 was visualized by gelatin zymography. Cytokine levels were determined by BioPlex assay. Intracellular ROS and free radical scavenging potential were measured by DCFH-DA and DPPH method, respectively. Xanthine oxidase activity was evaluated spectrophotometrically.

Results

The extract suppresses NF-κB activation by preventing Iκ-B phosphorylation and inhibits the phosphorylation of p38 and SAPK/JNK MAPKs. It down-regulates iNOS expression which manifests as a drastic decrease of NO production, inhibits MMP-9 activation, but does not affect COX-2 protein levels and reduces only slightly the released PGE₂. Secretion of IL-1β and Il-10 is greatly reduced, whereas suppression of TNF-α and GM-CSF production is less dramatic. The extract has strong free radical scavenging properties and exerts inhibitory effect on xanthine oxidase activity, which lowers the levels of intracellular ROS.

Conclusion

The study provides evidence for the anti-inflammatory potential of Clinopodium vulgare L. aqueous extract.     

iNOS/NO对结直肠肿瘤发生发展的影响

一氧化氮（nitric oxide,NO）具有广泛的生物学活性。近年来发现一氧化氮（NO）、一氧化氮合成酶（nitric oxide synthase-2,NOS-2）与结直肠肿瘤的发生、发展密切相关,它与环氧化酶（cy-cloxygenase-2,COX-2）之间存在复杂的调控机制。对NO清除剂、NOS抑制剂和释放NO的非甾体抗炎药的研究为结直肠肿瘤的防治提供了一个思路。     

Delayed expression of NADPH-diaphorase in rat brain after administration of the cholinotoxin AF64A

Administration of cholinotoxin etylcholine aziridinium (AF64A) into the brain selectively induces nonrever-sible cholinergic deficit. Wistar rats were injected intracerebroventricularly bilaterally with AF64A at doses of 1–3 nmol/ventricle. 28 days later the number of neurons survived was counted in dorsolateral, intermediate and medial groups of cells of the medial septum. AF64A induced a decrease in neuronal density and expression of cholineacetyl transferase at all doses used as well as in all regions studied. Brain sections were also stained for NADPH-diaphorase representing neuronal NO-synthase. Effects of AF64A on NADPH-diaphorase expression depended on the region studied. The number of NADPH-diaphorase-positive cells increased in the medial cellular group where more cholineacetly transferase-positive cells survived. In contrast, decrease in NADPH-diaphorase expression in the dorsolateral group of cells coincided with low level of cholineacetyltransferase-po-sitive neurons. The data presented suggest that in the AF64A-dependent model of neurodegeneration NO may play a neuroprotective function.     

两种艾灸法对二肾一夹型高血压大鼠血压和血管内皮细胞内分泌功能的影响

目的:应用两种艾灸疗法治疗两肾一夹肾血管性高血压大鼠(2K1C-RHR),评价这两种艾灸疗法的降压作用,并对其降压机理作初步的探讨。方法:建立2K1C-RHR模型,并将其随机分为六组:灸法Ⅰ组(百会、神阙、足三里)、灸法Ⅱ组(关元、涌泉、足三里)、卡托普利组、灸法Ⅰ+卡托普利组、灸法Ⅱ+卡托普利组、高血压对照组,另设正常对照组。经过10天治疗后,测量血压,并测定血浆中内皮素(ET)、一氧化氮(NO)。结果:高血压对照组的收缩压(SBP)、舒张压(DBP)明显高于正常对照组,各治疗组的SBP、DBP明显低于高血压对照组(P<0.01),各治疗组间则没有明显差异(P>0.05)。血浆NO含量各组间没有明显差异(P>0.05)。各治疗组与正常对照组的血浆中ET含量明显低于高血压对照组(P<0.01)。各治疗组ET/NO比值接近正常对照组(P>0.05)且明显低于高血压对照组(P>0.05)。结论:两种艾灸疗法有良好的降压作用,其降压机理与及纠正ET与NO的失衡状态有关。     

针刺对肾血管性高血压大鼠的降压效应

目的　探讨针刺对肾血管性高血压大鼠的降压作用及其神经 -内分泌调节机制。方法　采用二肾一夹法制备高血压大鼠模型 ,测定实验各组大鼠血浆内皮素 (ET)、一氧化氮 (NO)含量和心率变异性 (HRV)的动态变化。结果　与模型组相比 ,针刺组血压 (BL 2 3:19.1± 1.1k Pa,ST36:18.1± 2 .5 k Pa)及 ET水平明显下降 (P<0 .0 1) ,其中针刺肾俞组血浆 ET水平 (91.3± 9.45 pg/ ml)较足三里组 (10 8.48± 13.0 6pg/ ml)下降更明显 (P<0 .0 5 ) ;针刺治疗组血浆 NO水平 (BL 2 3:5 0 .3±12 .7μmol/ ml,ST 36:49.71± 8.49μm ol/ m l)显著升高 (P<0 .0 5 )。足三里组针刺后 L F/ HF比值与模型组无明显差异 ,而肾俞组 L F/ HF值 (0 .5 7± 0 .13)明显低于模型组及足三里组 (0 .86± 0 .2 5 )。结论　针刺能降低肾血管性高血压模型大鼠动脉血压 ,肾俞与足三里两穴无明显差异 ,其作用机制可能与自主神经系统、NO及 ET参与的神经内分泌调节作用相关。     

血管内皮型一氧化氮合酶基因Glu298 Asp多态性与老年脑梗死的相关性研究

目的　通过病例对照研究 ,了解中国老年人群血管内皮型一氧化氮合酶 (eNOS)基因目的　通过病例对照研究 ,了解中国老年人群血管内皮型一氧化氮合酶 (eNOS)基因Glu2 98Asp多态性的分布 ,分别探讨其与老年脑梗死及血脂以及一氧化氮等脑梗死危险因素的关系。 方法对门诊及住院中确诊的 4 0例老年脑梗死和 16 9例性别、年龄相匹配的老人 ,测量他们的身高、体重及座位血压 ,并测定他们的空腹血脂、空腹血糖 (FBS)及一氧化氮 (NO)等 ,应用聚合酶链反应 (PCR)和限制性片断长度多态性 (RFLP)检测eNOS基因Glu2 98Asp多态性。结果　脑梗死和对照组eNOS基因Glu2 98Asp多态性构成有显著性差异 (χ2 =4 31,P =0 0 38) ,脑梗死组Glu/Asp基因型高于对照组(32 5 %vs 17 8% ) ;脑梗死组 2 98Asp等位基因频率高于对照组 (16 2 5 %vs 8 9% ) ,但是两组等位基因频率的分布比较 ,没有显著性差异 (χ2 =3 81,P =0 0 5 1)。结论　eNOS基因Glu2 98Asp多态性在中国老年人群中存在 ,并且基因...     

Antioxidant and antiinflammatory activity of pine pollen extract in vitro

To determine the medicinal properties of pine pollen, the antioxidant and antiinflammatory activities of the ethanol extract of pine pollen extract (PPE) were investigated. PPE displayed a strong free radical scavenger activity on 1,1‐diphenyl‐2‐picrylhydrazyl radical and hydrogen peroxide. It was observed also that the antioxidant activity, measured by the ferric thiocyanate method, increased with the addition of PPE to the linoleic acid emulsion system. PPE was also found to inhibit significantly the amount of malondialdehyde and protein carbonyls formed from liver homogenate. Like the antioxidant activity, the reducing power of PPE was excellent. Thereafter, the study investigated the effects of PPE in modulating the production of pro‐inflammatory mediators in lipopolysaccharide (LPS)‐activated RAW 264.7 macrophages, and the effect of PPE on interleukin (IL)‐1β‐induced matrix metalloproteinases (MMPs) production and mitogen‐activated protein kinases (MAPKs) activation in the human synovial sarcoma cell line, SW982. PPE was found to inhibit the production of nitric oxide, tumor necrosis factor‐α, IL‐1 and IL‐6 in LPS‐activated macrophages. Treatment with PPE at 10 µg/mL significantly (p < 0.05) inhibited IL‐1β‐induced MMPs (MMP‐1 and ‐3) production in SW982 cells. IL‐1β‐induced JNK activation was inhibited by PPE (10 µg/mL), whereas p38 and ERK1/2 were not affected. These findings suggest that pine pollen is a potential antioxidant and beneficial for inflammatory conditions through down‐regulation of JNK and MMPs. Copyright © 2008 John Wiley & Sons, Ltd.     

L-精氨酸对胰腺炎相关性腹水诱导肾损伤的保护作用

目的 通过观察胰腺炎相关性腹水(PAAF)对正常肾脏的影响及L-精氨酸(L-arg)对其的作用,探讨重症急性胰腺炎时急性肾损伤的发病机制.方法 60只SD大鼠制成重症急性胰腺炎(SAP)模型并收集其腹水备用;60只SD大鼠制成消化道穿孔性腹膜炎(DPP)模型并收集其腹水(DAF)备用;将两组腹水分别注射至正常大鼠腹腔内诱导肾损伤以此建立PAAF组及DAF组;并观察L-精氨酸的作用.观察各组术后12 h的肾功能、血淀粉酶及病理学变化,同时检测肾细胞凋亡及诱生型一氧化氮合酶(iNOS)的表达情况.结果 血清BUN、Cr,PAAF组＞DAF组＞Larg组＞NS组;Amyl,DAF组＞PAAF组＞L-arg组＞NS组;但均低于SAP组;而血清NO及组织中iNOS的表达,NS组＜PAAF组＜DAF组＜SAP组＜L-arg组;肾细胞凋亡率及病理学改变,SAP组＞PAAF组＞DAF组＞L-arg组＞NS组.结论 PAAF可以诱导正常大鼠的肾细胞凋亡引起肾损伤,且该作用可以被小剂量的L-arg阻断.     

实验性自身免疫性脑脊髓炎巨噬细胞活性MR成像的研究进展

MR成像是监测多发性硬化(MS)病情及其演变、评价疗效的重要手段,实验性自身免疫性脑脊髓炎(EAE)是公认的研究人类多发性硬化的动物模型。常规MR扫描仅能分析MS和EAE炎症反应的继发性改变。巨噬细胞是MS和EAE炎症反应中重要的效应细胞,超微超顺磁性氧化铁(USPIO)粒子能被巨噬细胞摄取,MR成像能显示MS和EAE病灶内吞噬了USPIO的巨噬细胞,分析其炎症反应自身的信息,是动态观察巨噬细胞浸润过程、进一步探索MS免疫病理机制的有效手段。本文综述超微超顺磁性对比剂的特性及其MR成像在EAE和MS病理机制方面的研究进展,评价其发展前景。