Impact of matrine on inflammation related factors in rat intestinal microvascular endothelial cells

Ethnopharmacological relevance

Matrine (MT) is a main active ingredient of Sophora flavescens roots, which is used in Traditional Chinese Medicine (TCM) for the treatment of inflammations like enteritis, hepatitis and atopic dermatitis.

Aims of the study

Aim of the study is to gain insight into the effects of MT on nitric oxide (NO) release, intracellular NO production, and endothelial nitric oxide synthase (eNOS) level in second generation rat intestinal microvascular endothelial cells (RIMECs). Moreover, the effects of MT on soluble intercellular adhesion molecule-1 (sICAM-1), interleukin-6 (IL-6) and interleukin-8 (IL-8) production induced by lipopolysaccharide (LPS) in these cells were evaluated.

Material and Methods

Isolated and identified RIMECs cultures were exposed to different concentrations of matrine, and changes in extra- and intracellular NO concentrations were measured in dependance of time by Griess reaction or DAF-FM diacetate. Obtained cell cultures were solitude treated with lypopolysaccharide (LPS) or combined with MT to observe impacts on sICAM-1, IL-6 and IL-8 concentration in culture supernatants by ELISA.

Results

Matrine dose-dependently increased the concentration of NO in culture supernatant of RIMECs. Exposure of MT resulted in a steady intracellular NO increase pattern under different concentrations with different values and has an increasing effect on eNOS concentration at a long time exposure. Additionally, matrine reduced the increasing effect of LPS on the production of IL-6, IL-8, and sICAM-1 in RIMECs.

Conclusion

These results show that matrine may serve as a protective agent against tissue damage in inflammation by improving NO-dependent vasomotion and inhibiting inflammatory cytokines induced by LPS.     

小檗碱对大鼠肠黏膜微血管内皮细胞分泌一氧化氮的影响

目的研究小檗碱对体外培养大鼠肠黏膜微血管内皮细胞分泌一氧化氮（NO）的影响,探讨小檗碱治疗肠道疾病的药理作用机制。方法体外培养大鼠肠黏膜微血管内皮细胞,采用第Ⅷ因子相关抗原免疫荧光染色进行鉴定,在培养3、6、9、12h后,用比色法检测正常细胞与不同浓度小檗碱处理细胞上清液中NO含量,比较各组细胞NO的分泌水平。结果小檗碱处理组NO水平显著高于正常对照组,以5μg/mL剂量组作用最显著。结论促进内源性NO释放,介导肠黏膜微血管内皮依赖性舒张反应,改善肠道局部微循环,是小檗碱一个重要的药理作用机制。     

河南白头翁生物学特性的初步研究

目的：对河南地区野生白头翁生物学特性进行研究,为合理采收野生白头翁药材以及开发利用白头翁资源提供科学依据。方法：实地考察植物生长习性;高效液相包谱法测定野生白头翁中白头翁皂苷B4质量分数、2010年药典项下方法测定水分、灰分、酸不溶性灰分、醇浸物含量及其培养皿法测定种子千粒质量、发芽率、发芽势。结果：描述了白头翁的生物学特性,了解其生长发育规律及白头翁皂苷B4动态积累变化规律;种子发芽率、发芽势较低。结论：本实验为确定野生白头翁的适宜采收期以及保护开发利用野生白头翁资源提供了科学依据。     

白头翁中三萜皂苷类成分的药理研究进展

白头翁是一味用于治疗热毒血痢、温疟寒热、鼻衄、血痔的中药,其主要成分为五环三萜皂苷类,分属于齐墩果烷型和羽扇豆烷型。研究表明,白头翁三萜皂苷具有多种药理活性,包括抗肿瘤、抗炎、抗氧化、抗病毒、抗血吸虫、增强免疫等药理作用。该文综述白头翁三萜皂苷类成分的药理研究进展。     

Pharmacologic effects of lactones isolated from Pulsatilla alpina subsp. apiifolia

Two lactone compounds, protoanemonin and anemonin, were determined in the flowering aerial parts of P. alpina subsp. apiifolia. Anemonin is the primary compound responsible for the antipyretic activity and both anemonin and protoanemonin participate in the sedating effect.     

Effect of resveratrol on NF-kappaB activity in rat peritoneal macrophages

This study was to investigate the inhibitive effect of resveratrol (RESV) on nuclear factor kappa B (NF-kappaB) expression and activity induced by lipopolysaccharide (LPS) in rat peritoneal macrophages (PMA). Male Sprague-Dawley (SD) rats were randomly divided into 7 groups, including control group, LPS group and RESV I-V group. In the LPS group, PMA were incubated in DMEM containing LPS (10 microg/ml), whereas in control group, PMA were incubated in DMEM only. In the RESV I-V groups, PMA were incubated in DMEM containing LPS (10 microg/ml) and different concentrations of RESV. After 24 hours of incubation, NF-kappaB activity in PMA, and the levels of tumor necrosis factor alpha (TNF-alpha), interleukin-1 (IL-1) and nitric oxide (NO) in the culture medium were measured. In the concentrations of 1.25-5 microg/ml, RESV had a dose- dependent inhibitive effect on NF-kappaB activity in PMA as well as the expressions of TNF-alpha, IL-1 and NO in the culture medium contrasted with the LPS group. There was no significant difference in the levels of these pro-inflammatory factors between the groups of 5 microg/ml and 10 microg/ml RESV. In conclusion, RESV has the potential for the future application of preventing inflammatory diseases involving PMA.     

白头翁素诱导KB细胞凋亡的机制研究

目的：研究棉团铁线莲中的单体成分白头翁素诱导肿瘤细胞凋亡的机制。方法：细胞生长抑制试验测定白头翁素对KB细胞的IC50值;以DIOC6（3）为荧光探针,流式细胞术测定KB细胞的线粒体膜电位;以Caspse-9、-3酶活性检测试剂盒测定KB细胞中Caspse-9、-3的活性。结果：白头翁素对KB细胞生长抑制的IC50值为41.63±5.84μM,白头翁素可浓度依赖性地降低KB细胞的线粒体膜电位,白头翁素可时间依赖性地增强KB细胞的Caspse-9、-3活性。结论：白头翁素可诱导KB细胞发生线粒体途径的凋亡,这可能是其抗肿瘤作用机制之一。     

泻心汤有效组分及其配伍配比对大鼠腹腔巨噬细胞释放一氧化氮的影响

目的:观察泻心汤有效组分对巨噬细胞释放NO的影响,并对有效组分进行最佳配伍配比研究。方法:采用MTT法观察有效组分对巨噬细胞生长的影响,采用G riess法观察有效组分及其配伍配比对LPS诱导巨噬细胞产生NO的影响。结果:各组分在0.01~0.1 mg/ml剂量能明显抑制NO分泌,在LPS刺激细胞前1 h加入有效组分对NO的抑制作用弱于同时加药和LPS刺激后1 h加药;有效组分配伍后对NO分泌的抑制作用增强。结论:有效组分及其配伍配比均能抑制LPS诱导的巨噬细胞释放NO,且有效组分配伍后具有协同增效的特点。     

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.     

白头翁总皂苷在大鼠肠外翻试验中吸收特性考察

目的:考察白头翁总皂苷的肠吸收特性.方法:采用离体外翻肠囊模型研究白头翁总皂昔在不同肠段、不同药物浓度下的肠吸收特性,采用HPLC测定样品中指标成分常春藤皂苷元3-O-α-L-吡喃鼠李糖-(1→2)-[β-D-吡喃葡萄糖-(1→4)]-L-吡喃阿拉伯糖苷的质量浓度.结果:白头翁总皂苷指标成分在各个肠段的吸收无显著性差异,各肠段累积吸收量均随药物质量浓度的增加而增加.结论:白头翁总皂苷在大鼠肠道内小存在特殊的“吸收窗”,可能为被动扩散吸收.     

In vitro anti-inflammatory and anti-oxidative effects of Cinnamomum camphora extracts

Cinnamomum camphora Sieb (Lauraceae) has long been prescribed in traditional medicine for the treatment of inflammation-related diseases such as rheumatism, sprains, bronchitis and muscle pains. In this study, therefore, we aimed to investigate the inhibitory effects of Cinnamomum camphora on various inflammatory phenomena to explore its potential anti-inflammatory mechanisms under non-cytotoxic (less than 100 microg/ml) conditions. The total crude extract (100 microg/ml) prepared with 80% methanol (MeOH extract) and its fractions (100 microg/ml) obtained by solvent partition with hexane and ethyl acetate (EtOAc) significantly blocked the production of interleukin (IL)-1 beta, IL-6 and the tumor necrosis factor (TNF)-alpha from RAW264.7 cells stimulated by lipopolysaccharide (LPS) up to 20-70%. The hexane and EtOAc extracts (100 microg/ml) also inhibited nitric oxide (NO) production in LPS/interferon (IFN)-gamma-activated macrophages by 65%. The MeOH extract (100 microg/ml) as well as two fractions (100 microg/ml) prepared by solvent partition with n-butanol (BuOH) and EtOAc strongly suppressed the prostaglandin E(2) (PGE(2)) production in LPS/IFN-gamma-activated macrophages up to 70%. It is interesting to note that hexane, BuOH and EtOAc extracts (100 microg/ml) also inhibited the functional activation of beta1-integrins (CD29) assessed by U937 homotypic aggregation up to 70-80%. Furthermore, EtOAc and BuOH extracts displayed strong anti-oxidative activity with IC(50) values of 14 and 15 microM, respectively, when tested by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) and xanthine oxide (XO) assays. Taken together, these data suggest that the anti-inflammatory actions of Cinnamomum camphora may be due to the modulation of cytokine, NO and PGE(2) production and oxidative stress, and of the subfractions tested, the EtOAc extract may be further studied to isolate the active anti-inflammatory principles.     

祖师麻主要成分抗炎活性的体外研究

目的 观察祖师麻主要成分的抗炎活性.方法 分离提取祖师麻化学成分川木香醇D、左旋松脂酚、瑞香新素、西瑞香素、黄瑞香苷A、瑞香素、黄瑞香苷B,采用CCK-8实验进行细胞毒性评价.将细胞按随机数字表法分为对照组、模型组和化合物组.对照组和模型组各加入50μl培养液,化合物组分别加入50.00、25.00、12.50、6.25、3.12μg/ml西瑞香素、黄瑞香苷A、黄瑞香苷B溶液,模型组和化合物组再加入4μg/ml脂多糖50μl,刺激24 h.采用Griess试剂法测定NO释放量,采用ELISA法检测TNF-α分泌量.结果 西瑞香素、黄瑞香苷A、黄瑞香苷B在50μg/ml下对小鼠巨噬细胞RAW264.7细胞具有较弱的细胞毒性,而其他化合物细胞毒性作用较明显.与模型组比较,12.50、25.00、50.00μg/ml黄瑞香苷B可抑制RAW264.7细胞NO[(271.86±20.92)%、(256.48±20.92)%、(199.31±15.16)%比(358.62±28.64)%]、TNF-α[(647.87±115.79)%、(618.42±87.52)%、(588.33±87.94)%比(1035.06±58.29)%]生成(P<0.05或P<0.01);25.00、50.00μg/ml黄瑞香苷A可抑制RAW264.7细胞NO[(234.99±34.28)%、(167.36±25.76)%比(358.62±28.64)%]、TNF-α[(691.76±60.37)%、(534.01±41.60)%比(1035.06±58.29)%]生成(P<0.05或P<0.01);12.50、25.00、50.00μg/ml西瑞香素可抑制RAW264.7细胞NO[(283.89±36.69)%、(243.08±48.19)%、(225.92±33.67)%比(358.62±28.64)%]、TNF-α[(713.77±121.96)%、(670.62±18.70)%、(599.62±68.62)%比(1035.06±58.29)%]生成(P<0.05或P<0.01).结论 黄瑞香苷A、黄瑞香苷B和西瑞香素具有抗炎作用,可降低脂多糖诱导的RAW264.7细胞NO释放量和TNF-α分泌量,发挥抗炎作用.     

Nitric oxide inhibitory substances from the rhizomes of Dioscorea membranacea

Thai medicinal plants locally known as Hua-Khao-Yen were examined for their inhibitory activities against lipopolysaccharide (LPS) induced nitric oxide (NO) production in RAW 264.7 cell lines. Among the plant species studied, an ethanolic extract of Dioscorea membranacea exhibited the most potent inhibitory activity, with an IC(50) value of 23.6 microg/ml. From this extract, eight compounds [two naphthofuranoxepins (1, 2), one phenanthraquinone (3), three steroids (4-6) and two steroidal saponins (7, 8)] were isolated and further investigated for their inhibitory properties of NO production. It was found that diosgenin-3-O-alpha-L-rhamnosyl (1-->2)-beta-D-glucopyranoside (7) possessed the highest activity (IC(50)=3.5 microM), followed by dioscoreanone (3, IC(50)=9.8 microM) and dioscorealide B (2, IC(50)=24.9 microM). Regarding structural requirements of diosgenin derivatives for NO production inhibitory activity, compound 7 which has a rhamnoglucosyl moiety at C-3 exhibited much higher activity than compounds that have either a diglucosyl substitution (8) or its aglycone (9); whereas, hydroxyl substitution at position 8 of naphthofuranoxepin derivatives conferred higher activity than the methoxyl group. It is concluded that diosgenin-3-O-alpha-L-rhamnosyl (1-->2)-beta-D-glucopyranoside (7), dioscoreanone (3) and dioscorealide B (2) are active principles for NO inhibitory activity of Dioscorea membranacea. Compounds 1-3 were also tested for the inhibitory effect on LPS-induced TNF-alpha release in RAW 264.7 cells. The result revealed that 3 possessed potent activity against TNF-alpha release with an IC(50) value of 17.6 microM, whereas, 1 and 2 exhibited mild activity. The present study may support the use of Dioscorea membranacea by Thai traditional doctors for treatment of the inflammatory diseases.     

电针对局灶性脑缺血再灌注大鼠外周血中可溶性细胞间粘附分子-1和内皮素-1的影响

目的:探讨电针对大脑中动脉缺血再灌注模型大鼠外周血中可溶性细胞间粘附分子-1(sICAM-1)、内皮素-1(ET-1)含量的影响。方法:SD大鼠40只,随机分为正常对照组、假手术组、模型组、电针组,采用大脑中动脉线栓法制备局灶性脑缺血再灌注模型,于缺血再灌注24 h后摘除眼球采血,分别应用酶联免疫吸附法、放射免疫法观察脑缺血再灌注及电针对大鼠外周血中sICAM-1、ET-1含量的影响。结果:脑缺血再灌注后大鼠外周血中sICAM-1、ET-1的含量增高(模型组与正常组、假手术组比较P<0.01);电针可降低大鼠外周血中sICAM-1、ET-1的含量(电针组与模型组比较P<0.05)。结论:早期针刺治疗对脑缺血损害有效防治作用是通过改善脑血管内皮细胞功能而实现的。     

Inhibition of lipopolysaccharide and interferon-gamma-induced expression of inducible nitric oxide synthase and tumor necrosis factor-alpha by Lithospermi radix in mouse peritoneal macrophages

Lithospermi radix (LR, root of Lithospermum erythrorhizon Siebold. et Zuccarinii) has been used to treat various conditions, such as septic shock, eczema and burns. In this study, the effect of LR on lipopolysaccharide (LPS) and recombinant interferon-gamma (rIFN-gamma)-induced production of nitric oxide (NO) and tumor necrosis factor (TNF)-alpha were examined using mouse peritoneal macrophages. At 0.01-1 mg/ml, LR inhibited the LPS/rIFN-gamma-induced expression of inducible nitric oxide synthase (iNOS) and TNF-alpha release. To clarify the mechanism involved, the effect of LR on the activation of nuclear factor (NF)-kappaB was examined. The LPS/rIFN-gamma-induced activation of NF-kappaB was almost completely blocked by LR at 1mg/ml without cytotoxicity. These findings demonstrate that the inhibition of the LPS/rIFN-gamma-induced production of NO and TNF-alpha by LR involves the inhibition of NF-kappaB activation.     

Supercritical carbon dioxide extract exhibits enhanced antioxidant and anti-inflammatory activities of Physalis peruviana

Physalis peruviana L. (PP) is a medicinal herb widely used in folk medicine. In this study, supercritical carbon dioxide (SFE-CO2) method was employed to obtain three different PP extracts, namely SCEPP-0, SCEPP-4 and SCEPP-5. The total flavonoid and phenol concentrations, as well as antioxidant and anti-inflammatory activities of these extracts were analyzed and compared with aqueous and ethanolic PP extracts. Among all the extracts tested, SCEPP-5 demonstrated the highest total flavonoid (234.63+/-9.61 mg/g) and phenol (90.80+/-2.21 mg/g) contents. At concentrations 0.1-30 microg/ml, SCEPP-5 also demonstrated the strongest superoxide anion scavenging activity and xanthine oxidase inhibitory effect. At 30 microg/ml, SCEPP-5 significantly prevented lipopolysaccharide (LPS; 1 microg/ml)-induced cell cytotoxicity in murine macrophage (Raw 264.7) cells. At 10-50 microg/ml, it also significantly inhibited LPS-induced NO release and PGE2 formation in a dose-dependent pattern. SCEPP-5 at 30 microg/ml remarkably blocked the LPS induction of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression. Taken together, these results suggest that SCEPP-5, an extract of SFE-CO2, displayed the strongest antioxidant and anti-inflammatory activities as compared to other extracts. Its protection against LPS-induced inflammation could be through the inhibition of iNOS and COX-2 expression.