白花蛇舌草的化学成分研究

为研究白花蛇舌草(Hedyotis diffusa)的化学成分,采用硅胶柱色谱、制备HPLC色谱等方法,从白花蛇舌草全草的70%乙醇提取物中分离得到9个化合物。根据理化性质及其波谱或光谱数据,结构分别鉴定为：豆甾醇(1)、β-谷甾醇(2)、rubiadin(3)、2,6-二羟基-1-甲氧基蒽醌(4)、β-谷甾醇-3-O-β-D-葡萄糖苷(5)、对香豆酸(6)、山奈酚(7)、槲皮素(8)和2-羟基-3-甲氧基-7-甲基蒽醌(9)。化合物4为新化合物;氯仿提取部位的Fr.4、Fr.4-2及Fr.4-4流分具有较强体外抗人子宫内膜癌Ishikawa细胞活性,其IC₅₀值分别为52.8、78.1和27.5μg/mL。     

Protective Antioxidant Effects of Amentoflavone and Total Flavonoids from Hedyotis diffusa on H2O2‐Induced HL‐O2 Cells through ASK1/p38 MAPK Pathway

In this study, total flavonoids and total triterpenoid acid were extracted with ethyl acetate from Hedyotis diffusa Willd, and hepatoprotective activities of them and five compounds from total flavonoids against H₂O₂ induced hepatocyte damage on HL‐02 cells were determined. In particular, amentoflavone and total flavonoids had influence on the leakage of ALT, AST, LDH, the activities of SOD and the content of MDA. They effectively reduced the loss of MMP, the release of Cyt C, and then inhibited activation of caspase‐3/caspase‐9 cascade in hepatotoxic cells. The contents of ROS were significantly reduced to inhibit p38 in amentoflavone and flavonoids groups which decreased ASK1 and p‐p38 levels through increasing thioredoxin Trx1 and reductase TrxR1. These results suggesting that the antioxidant protection of amentoflavone and flavonoids might be reducing ROS to inhibit the H₂O₂‐induced upstream of pathway via increasing levels of Trx1 and TrxR1, which were pivotal in blocking the down streaming effectors of ASK1/p38 MAPK pathway and alleviating hepatotoxicity.     

Development of microsatellite markers in heterostylous Hedyotis chrysotricha (Rubiaceae)

? Premise of the study: Microsatellite primers were developed for a heterostylous herb, Hedyotis chrysotricha to investigate the effect of habitat fragmentation on its genetic diversity and population structure. ? Methods and Results: Twelve primer sets were developed and their polymorphisms were tested on 47 individuals from two island populations of H. chrysotricha in Thousand Island Lake of China. The number of alleles per locus ranged from five to 10, with an average of seven alleles. Expected heterozygosity per locus ranged from 0.284 to 0.821 and observed heterozygosity ranged from 0.191 to 0.851. ? Conclusions: We showed that all of the 12 microsatellite markers developed for H. chrysotricha are polymorphic within populations, which should provide a powerful tool for assessing population structure and genetic diversity across fragmented and continuous populations, and for studying the genetic effects of habitat fragmentation on this species.     

耳草属(HedyotisL.)的起源及地理分布

依据耳草属的系统学,古地理学和细胞染色体资料分析和推论,耳草属植物的起源地点在冈瓦纳古陆,很可能在古陆辽阔的东北部地区。耳草属植物的起源时间不应晚于侏罗纪,有可能在三叠纪就已经出现,并在侏罗纪得到广泛的传播,其迁移的路线有四条,即从起源中心向东经上耳其,伊朗进入东南亚地区,向东南通过古南大陆向印度板块和徜大利亚扩散,向北进入北美地区,向西南进入南美洲地区,随着冈瓦纳古陆的分离,印度板块向北漂移以及澳大利亚与古南大陆的分离,植物迁移与扩散的速度受到了制约,耳草属植物现代分布格局形成的原因在于传播途径的隔断和第四纪冰川的作用。     

白花蛇舌草的抗肿瘤活性和初步化学研究

我们在美国国立肿瘤研究所对白花蛇舌草 (Hedyotisdiffusa)的乙醇提取物及不同的溶剂萃取部位进行了肿瘤细胞株的细胞毒活性筛选 ,发现CH3 OH∶H2 O(9∶1)部位有一定的细胞毒活性 ,特别是对乳腺癌细胞株有中等强度活性。因此我们对该部位进行了初步化学成分研究 ,从中分离得到三个已知化合物 ,即豆甾醇 5 ,2 2 二烯 3β ,7α 二醇 (Stigmasta 5 ,2 2 diene 3β ,7α diol,1) ,豆甾醇 5 ,2 2 二烯 3β ,7β 二醇 (Stigmasta 5 ,2 2 diene 3β ,7β diol,2 )和熊果酸 (Ursolicacid ,3)。其中化合物 (1)和化合物 (2 )系首次从该植物中分离得到 ,同时文献报道这类甾醇类化合物有一定的细胞毒活性。进一步的活性和化学研究仍在进行。     

白花蛇舌草对Ⅱ型胶原蛋白诱导的大鼠类风湿性关节炎的治疗作用

目的：探讨白花蛇舌草（HD）对Ⅱ型胶原蛋白诱导的大鼠类风湿性关节炎（CIA）的治疗作用。方法：取SD大鼠60只随机分为对照组10只、造模组50只,造模组于背部皮内注射Ⅱ型胶原乳剂建立CIA大鼠模型,采用大鼠关节炎评分法评价其模型是否复制成功。造模组再随机分为模型组、雷公藤多苷（GTW）6 mg/kg组、白花蛇舌草3、6、12 g/kg组,每组10只,每天灌胃给予相应的试剂。每周观察关节炎指数和痛阈,于连续干预28 d后,各组大鼠眼眶静脉取血,观察白细胞介素1β（IL-lβ）、肿瘤坏死因子-α（TNF-α）、前列腺素E₂（PGE₂）、核因子κB受体活化因子配体（RANKL）及护骨素（OPG）水平。结果：与对照组比较,模型组大鼠关节炎指数,血清IL-lβ、TNF-α、PGE₂、RANKL、OPG及RANKL/OPG水平均明显升高（P<0.05）,光照阈值明显降低（P<0.05）;与模型组比较,雷公藤多苷组、白花蛇舌草低、中、高剂量组关节炎指数,血清IL-lβ、TNF-α、PGE₂、RANKL、OPG及RANKL/OPG水平均明显降低（P<0.05）,光照阈值明显升高（P<0.05）。结论：白花蛇舌草可明显降低CIA模型大鼠关节炎指数,升高痛域阈值,下调IL-lβ、TNF-α、PGE₂、RANKL、OPG水平,从而有效的防治CIA。     

Application of DNA barcodes in Hedyotis L. (Spermacoceae,Rubiaceae)

The potential application of DNA barcodes of plastid (matK, trnH–psbA, petD, and rbcL) and nuclear (internal transcribed spacer (ITS) of rDNA) DNA regions was investigated for 25 Hedyotis taxa. The ITS showed the best species discrimination by resolving 23 of the species as exclusive lineages with no shared alleles between any of the 24 distinct species (H. assimilis and H. mellii are not supported as distinct species based on our molecular and morphological data). Conversely, rbcL performed the worst and only resolved 10 of the species as exclusive lineages, and 10 species with shared alleles. Using ITS has the advantage of high PCR amplification success and it provides good intra- and interspecific variation distribution patterns. The most powerful plastid markers were petD and trnH–psbA, but we could amplify and sequence trnH–psbA for only 83% of the accessions sampled. Combination of ITS and petD performed extremely well, with all 24 of the distinct species resolved as exclusive lineages and no shared alleles between any of the distinct species. We therefore recommend ITS, or a combination of ITS and petD, as the standard DNA barcode in Hedyotis, but acknowledge that there are no shared alleles between distinct species for matK and rbcL combined.     

闭花耳草的化学成分研究

从闭花耳草(Hedyotis cryptantha )全草中分离得到11个化合物,经波谱数据分析鉴定为车叶草苷(1),车叶草苷酸(2),车叶草苷酸甲酯(3),车叶草酸乙酯(4),3,4-二氢-3-甲氧基车叶草苷(5),山奈酚(6),kaempferol 3,7-di-O -β-D-glucoside(7),kaempferol 3-O -β-D-galactopyranosyl-(1→3)-β-D-galactopyranoside(8),乌索酸(9),豆甾醇(10),β-谷甾醇(11)。所有化合物均为首次从闭花耳草植物中分离得到,其中化合物7和8首次从耳草属(Hedyotis )植物中分离得到。     

Hedyotis koiana R. J. Wang,a new species of Rubiaceae from China

Hedyotis koiana R. J. Wang is described as a new species from Guangdong, Hunan, Jiangxi, Guangxi, Fujian, and Hainan provinces of China. It is most similar to H. herbacea L. and H. pinifolia Wall. ex G. Don. because they all have narrow leaves and their capsules are loculicidally dehiscent at the top, but it differs from H. herbacea by having termi- nal or upper axillary cyme and narrow leaves, and it can be distinguished from H. pinifolia by having long flowering pedicles, sparse inflorescence, and adaxially glabrous corolla.     

不同提取方法对豫产白花蛇舌草多糖及抗氧化活性的影响

为评价不同提取方法对河南地区白花蛇舌草粗多糖的得率、总碳水化合物含量、杂质含量和抗氧化活性的影响,本实验分别以传统热水提取法、超声波辅助提取法、纤维素酶提取法从该药材中提取多糖。同时测定了粗多糖中总碳水化合物含量、杂质含量(总蛋白和总酚含量)以及羟基自由基清除能力、DPPH自由基清能力、还原力等抗氧化能力。实验结果显示三种提取法所得粗多糖相对于原药材的得率分别为6.77±0.09%、6.37±0.01%和4.81±0.90%;粗多糖中的总碳水化合物含量分别为289.14±1.33、246.83±2.33和278.92±2.92mg/g。粗多糖中总蛋白含量分别为28.59±2.21、23.26±2.43和4.96±0.18mg/g,总酚类成分含量分别为5.9±0.08、5.31±0.40和2.82±0.07mg/g。抗氧化活性实验结果表明,三种粗多糖的羟基自由基清除能力呈现出浓度依赖性,且均在3.0mg/mL时达到最大,对应最大清除率分别为89.13%、85.87%和74.71%。另外,三种粗多糖的DPPH自由基清除率分别在0.75~3.0mg/mL的浓度区间内达到最大并保持稳定,对应清除率的稳定区间分别为77.02%~77.90%、77.06%~77.96%和83.16%~85.73%。综上所述,以上三种不同提取方法对白花蛇舌草多糖的品质影响较大,可以为豫产白花蛇舌草的品质鉴别、多糖质量控制和开发利用提供依据。