中药影响实验性自身免疫性脑脊髓炎CD4+T细胞亚群分化的研究进展

实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)是由CD4⁺T细胞介导的中枢神经系统脱髓鞘性疾病,是国际公认研究多发性硬化(multiple sclerosis,MS)的动物模型。CD4⁺T细胞作为EAE模型的主要免疫应答细胞,存在多个亚群,可分泌多种细胞因子,参与病情进展。CD4⁺T细胞增殖分化及功能紊乱与EAE发病机制密切相关。目前临床尚无治疗MS的有效药物,多以对症应用皮质类固醇激素为主,但不良反应严重,且易产生药物依赖。近年来中药因作用温和、药物安全性较高、成本低、耐受性好等优点逐渐走进大众视野,针对中药对EAE中CD4⁺T细胞亚群分化的影响展开论述,为临床用药提供更多理论基础。     

The Effects of Pre‐Exercise Ginger Supplementation on Muscle Damage and Delayed Onset Muscle Soreness

Ginger possesses analgesic and pharmacological properties mimicking non‐steroidal antiinflammatory drugs. We aimed to determine if ginger supplementation is efficacious for attenuating muscle damage and delayed onset muscle soreness (DOMS) following high‐intensity resistance exercise. Following a 5‐day supplementation period of placebo or 4 g ginger (randomized groups), 20 non‐weight trained participants performed a high‐intensity elbow flexor eccentric exercise protocol to induce muscle damage. Markers associated with muscle damage and DOMS were repeatedly measured before supplementation and for 4 days following the exercise protocol. Repeated measures analysis of variance revealed one repetition maximum lift decreased significantly 24 h post‐exercise in both groups (p < 0.005), improved 48 h post‐exercise only in the ginger group (p = 0.002), and improved at 72 (p = 0.021) and 96 h (p = 0.044) only in the placebo group. Blood creatine kinase significantly increased for both groups (p = 0.015) but continued to increase only in the ginger group 72 (p = 0.006) and 96 h (p = 0.027) post‐exercise. Visual analog scale of pain was significantly elevated following eccentric exercise (p < 0.001) and was not influenced by ginger. In conclusion, 4 g of ginger supplementation may be used to accelerate recovery of muscle strength following intense exercise but does not influence indicators of muscle damage or DOMS. Copyright © 2015 John Wiley & Sons, Ltd.     

姜辣素对顺铂致大鼠异嗜模型行为的作用

目的研究姜辣素对顺铂所致大鼠异嗜高岭土模型的影响并观察外周和中枢c-fos蛋白的表达。方法大鼠随机分成正常对照组、模型组、昂丹司琼+地塞米松组、阿瑞吡坦+地塞米松组、姜辣素组及姜辣素+地塞米松组,24 h及72 h记录各组摄食量及食高岭土量。免疫组化方法观察脑和回肠中c-fos蛋白的表达。结果 24 h内,姜辣素组、姜辣素+地塞米松组摄食量增加,啃食高岭土量减少,姜辣素组与姜辣素+地塞米松组相比摄食量增加(P<0.05);48～72h内,姜辣素组食高岭土量减少(P<0.05)。姜辣素组、姜辣素+地塞米松组c-fos阳性表达颗粒的灰度值明显高于模型组(P<0.05)。结论姜辣素可促进摄食并可抑制顺铂所致的大鼠异嗜高岭土行为,地塞米松对姜辣素无辅助作用。     

生姜中姜酚类活性成分的抗肿瘤作用及其机制

目的 从生姜中提取、分离和鉴定姜酚类物质,初步研究其抗肿瘤作用的分子机制.方法 采用CO2超临界流体对生姜原料进行提取,通过硅胶、RP-C18、Sephadex LH-20和HPLC等色谱方法进行分离和纯化,进而借助MS和NMR等光谱方法鉴定化合物结构,然后采用MTT法检测化合物对不同肿瘤细胞活性的抑制作用,采用流式细胞术及Western blot检测其抑制肿瘤细胞活性的变化.结果 从生姜中提取分离得到5个姜酚类化合物,分别为4,6,8,10-姜酚和5’-羟基-6-姜酚.其中8-姜酚和10-姜酚对多种肿瘤细胞的活性均有较好的抑制作用,尤以10-姜酚作用最强,给药72 h后其对乳腺癌MDA-MB-231、MCF-7细胞的IC50分别为(25.80±1.39)、(35.29±2.70) μmol/L.流式细胞术检测发现8-姜酚和10-姜酚可导致乳腺癌细胞的G1期阻滞,其中10-姜酚作用于MDA-MB-231和MCF-7细胞24 h后G1期细胞的百分比分别为(66.73±2.93)％、(66.59±2.49)％,相对于对照组的(47.39±1.97)％和(49.17±3.52)％,差异有统计学意义(P＜0.01).Western blot检测结果显示,8-姜酚和10-姜酚可下调MDA-MB-231和MCF-7细胞中G1期相关蛋白Cyclin D1和CDK4的表达;并可降低MAPK信号通路中ERK的磷酸化水平,增强P38的磷酸化水平.结论 从生姜中提取出的8-姜酚和10-姜酚具有明显的抑制肿瘤细胞活性的作用,其机制可能与其影响MAPK通路中ERK、P38磷酸化水平,导致细胞G1期阻滞有关.     

6-gingerol prevents patulin-induced genotoxicity in HepG2 cells

Patulin (PAT) is a mycotoxin produced by several Penicillium, Aspergillus and Byssochlamys species. Since PAT is a potent genotoxic compound, and PAT contamination is common in fruits and fruit products, the search for newer, better agents for protection against genotoxicity of PAT is required. In this study, the chemoprotective effect of 6-gingerol against PAT-induced genotoxicity in HepG2 cells was investigated. The comet assay and micronucleus test (MNT) were used to monitor genotoxic effects. To further elucidate the underlying mechanisms, the intracellular generation of reactive oxygen species (ROS) and level of reduced glutathione (GSH) were tested. In addition, the level of oxidative DNA damage was evaluated by immunocytochemical analysis of 8-hydroxydeoxyguanosine (8-OHdG). The results showed that 6-gingerol significantly reduced the DNA strand breaks and micronuclei formation caused by PAT. Moreover, 6-gingerol effectively suppressed PAT-induced intracellular ROS formation and 8-OHdG level. The GSH depletion induced by PAT in HepG2 cells was also attenuated by 6-gingerol pretreatment. These findings suggest that 6-gingerol has a strong protective ability against the genotoxicity caused by PAT, and the antioxidant activity of 6-gingerol may play an important part in attenuating the genotoxicity of PAT.     

姜酚肟对谷氨酸诱导PC12细胞损伤的保护作用

 【目的】探讨姜酚肟对谷氨酸(glutamate, Glu)诱导PC12细胞损伤的保护作用。【方法】体外培养PC12细胞，建立谷氨酸诱导PC12细胞损伤的模型；采用MTT法检测细胞活力，流式细胞术和Hoechst 33258 DNA 染色法检测细胞凋亡。【结果】经5 mmol/L的谷氨酸处理24 h后，PC12细胞活力比对照组降低，仅为对照组的58.3﹪。在一定浓度范围内，姜酚肟能保护PC12 细胞免受谷氨酸(5 mmol/L)的影响，但较高浓度时对细胞有一定的毒性。经不同浓度姜酚肟(3.125、6.25、12.5、25 μg/mL)预处理后，细胞活力明显提高，其保护作用随浓度降低而升高，当浓度为6.25 μg/mL时效果最好，使PC12细胞的存活率达到75.2﹪（P<0.01），浓度为3.125 μg/mL 时，细胞存活率降为70.2﹪(P<0.01)。谷氨酸(5 mmol/L)能诱导PC12细胞凋亡，处理24 h 后细胞凋亡率为20.1﹪, 经姜酚肟(3.125、6.25、12.5、25 μg/ml )预处理后，细胞凋亡率明显降低，分别为3.5﹪(P<0.01), 2.8﹪(P<0.01), 9.6﹪(P<0.01), 17.7﹪(P<0.05)。【结论】谷氨酸能诱导PC12细胞凋亡，较低浓度的姜酚肟能有效保护PC12细胞免受谷氨酸诱导的细胞损伤，其中6.25 μg/ml的姜酚肟效果最好。     

6‐Gingerol‐rich fraction from Zingiber officinale ameliorates carbendazim‐induced endocrine disruption and toxicity in testes and epididymis of rats

This study evaluated the protective effects of 6‐gingerol‐rich fraction (6‐GRF) from Zingiber officinale on carbendazim (CBZ)‐induced reproductive toxicity in rats. Adult male rats were treated with either CBZ (50 mg/kg) alone or in combination with 6‐GRF (50, 100 and 200 mg/kg) for 14 consecutive days. Gas chromatography–mass spectrometry (GCMS) analysis revealed that 6‐GRF consists of ten bioactive chemical components with 6‐gingerol being the most abundant (30.76%). Administration of 6‐GRF significantly (p < .05) prevented CBZ‐mediated increase in absolute and relative testes weights as well as restored the sperm quantity and quality in the treated rats to near control. In testes and epididymis, 6‐GRF significantly abolished CBZ‐mediated increase in oxidative damage as well as augmented antioxidant enzymes activities and glutathione level in the treated rats. Moreover, CBZ administration alone significantly decreased plasma levels of testosterone, thyrotropin, triiodothyronine and tetraiodothyronine, whereas follicle‐stimulating hormone was significantly elevated without affecting luteinising hormone and prolactin levels when compared with the control. Conversely, 6‐GRF ameliorated the disruption in the hormonal levels and restored their levels to near normalcy in CBZ‐treated rats. Collectively, 6‐GRF inhibited the adverse effects of CBZ on the antioxidant defence systems, hormonal balance and histology of the testes and epididymis in rats.     

Species differences in the prokinetic effects of ginger

This study describes the prokinetic actions of the aqueous extract of ginger (Zingiber officinale). Ginger extract (Zo.Cr), which tested positive for saponins, terpenes, phenols, flavonoids and alkaloids, showed a spasmogenic effect in isolated guinea-pig ileum with 8–50 times more potency than in rabbit jejunum and ileum and rat stomach fundus and ileum. Spasmogenicity in all the gut preparations except in guinea-pig ileum was atropine-sensitive. Zo.Cr exhibited a stimulant effect in vivo in mice and enhanced the intestinal transit of charcoal meal. A spasmolytic effect, mediated via Ca^2?+? antagonist activity, was also exhibited by Zo.Cr, reflected in terms of inhibition of spontaneous contractions, K⁺ (80 mM)-induced contractions and displacement of Ca^2?+? dose–response curves. The ginger pure compounds (6-shogaol, 6-gingerol, 8-gingerol and 10-gingerol) also exhibited a spasmolytic activity, which reduced with the increasing size of the side chain in their chemical structures. The study showed that the aqueous extract of ginger exhibits species-specific spasmogenicity in gut tissues of rabbit and rat (muscarinic-type) while through an uncharacterized pathway in guinea-pig ileum, along with a dormant relaxant effect, mediated via the blockade of voltage-dependent Ca^2?+? channels.     

基于UPLC特征指纹图谱和指标成分定量测定研究炮姜的炮制工艺

目的 从UPLC特征指纹图谱和6-姜酚、8-姜酚、10-姜酚量的角度,选择炮姜的最佳炮制温度和炮制时间.方法 采用砂烫法,选取180、185、190、195、200℃,每个温度下分别炒制6、7、8 min,共得到15份样品;采用超高效液相色谱(UPLC)法分析各样品指纹图谱相似度;同时采用UPLC法测定不同炮制品中6-姜酚、8-姜酚、10-姜酚的量.结果 195℃、7 min和195℃、8 min 2份样品的相似度最大,分别为0.995和0.994;6-姜酚、8-姜酚、10-姜酚的总量最低,分别为5.792、5.412 mg/g.结论 综合指纹图谱相似度和6-姜酚、8-姜酚、10-姜酚量的测定结果以及炮制品外观,确定炮姜砂烫炮制以195℃、7 min为宜.     

高效液相色谱法测定姜酚胶丸中6-姜酚的含量

目的建立测定姜酚胶丸中6-姜酚含量的HPLC法。方法色谱柱为ODSC18柱（4．6mm×250rnm,5μm）,流动相：甲醇-水（65：35,v／v）,流速：1．0mL·min-1检测波长：280nm,柱温：室温。结果6-姜酚在40-640μg·mL。与峰面积呈良好线性关系,r=0．9998;平均回收率为99．8％,RSD—1．29％。结论该方法简便、准确、可靠,适用于姜酚胶丸的定量检测。