冬虫夏草用药部位描述的商榷

对国家药典所载冬虫夏草用药部位的描述进行商榷。建议描述为:冬虫夏草为麦角菌科真菌冬虫夏草菌Cordyceps sinensis(Berk.)Sacc.侵染了蝙蝠蛾科昆虫幼虫而形成的成熟菌体及幼虫尸体的干燥复合体,对冬虫夏草的基原描述提出新的观点,为其描述的准确性提供新的参考。     

冬虫夏草可溶性蛋白质多样性与产地关联度初探

采用bradford法,以双向电泳技术对不同产地、不同初加工方式的冬虫夏草中可溶性蛋白质的含量、种类进行分析,发现冬虫夏草不同加工方法后可溶性蛋白差异明显.不同产地冬虫夏草中可溶性蛋白质多样性差异也较为明显.初步表明可溶性蛋白质含量、多样性指标与冬虫夏草加工方式、产地有一定的相关性.     

冬虫夏草与其混淆品的性状及显微鉴别研究

目的:提供一种既简单又方便的方法,来区分正品冬虫夏草及其混淆品.方法:采用性状鉴别法,观察虫草类的虫体及子座,以及粉末显微鉴别法.结果:性状鉴别方面,冬虫夏草的正品及混淆品可通过子座的有无或形状,菌膜、虫体气门的排列方式和虫体颜色等特征区分.显微鉴别方面,则可从虫体体壁细胞的表面纹理,及背、足部短刚毛的有无、长短、形状和排列方式等各方面鉴别其品种.结论:该方法简单易行,可用于鉴别冬虫夏草的正品及混淆品.     

人工冬虫夏草联合糖皮质激素对大鼠肺纤维化的干预作用

目的:探讨人工冬虫夏草(虫草菌液)联合糖皮质激素对博莱霉素诱导的大鼠肺纤维化的干预作用.方法:50只SD大鼠,随机分为5组:正常对照组、模型组、虫草菌液组、强的松组、虫草菌液联合强的松组(联合组).采用气管内注射博莱霉素建立模型,次日起分别给药灌胃治疗,正常组和模型组给予生理盐水.第28天处死动物,肺组织标本行HE染色及MASSON染色进行病理形态学观察、Ashcroft纤维化评分,碱水解法检测肺组织HYP含量,免疫组化检测肺组织CTGF蛋白表达,酶联免疫法检测肺泡灌洗液上清TGF-β1含量.结果:与模型组相比,虫草菌液、强的松组能够降低肺组织纤维化评分,减少肺组织胶原沉积、HYP含量,下调肺组织CTGF蛋白表达,降低BLAF中TGF-的含量;联合组分别与虫草菌液组、强的松组比较,能够进一步减少肺纤维化评分、胶原面积、HYP含量,进一步下调CTGF蛋白、TGF-蛋白表达.结论:虫草菌液、强的松对博莱霉素诱导的大鼠肺纤维化有治疗作用,且虫草菌液联合强的松能产生协同治疗效果.     

Comparisons on enhancing the immunity of fresh and dry Cordyceps militaris in vivo and in vitro

Ethnopharmacological relevance

The immunomodulatory capacities of fresh Cordyceps militaris (FCM) and dry Cordyceps militaris (DCM) were compared.

Materials and methods

In vivo immunomodulatory assay, different doses of FCM and DCM were orally administrated over a period of 15 days in a cyclophosphamide (CY) induced immunosuppression mice; in vitro testing, the spleen cells were extracted from healthy mice and treaded with CY, then cultured with different dose of FCM or DCM; the contents of Cordyceps militaris polysaccharide (CMP), cordycepin, adenosine, total polyphenol (TP) and total flavonoids (TF) in FCM and DCM were measured.

Results

Our studys indicated that, FCM was significantly stronger than DCM on increasing the spleen and thymus indexes, spleen lymphocyte activity, macrophage function, and promoting the levels of IL-2, IFN-γ in vivo and in vitro. The contents of immunomodulatory CMP and TF in FCM were markedly higher than in DCM.

Conclusion

All these results suggested that FCM was superior to DCM on enhancing immunity.     

白木香叶化学成分的研究

目的:研究白木香叶的化学成分。方法:应用溶剂法和色谱法分离纯化化合物,利用谱学技术鉴定化合物结构。结果:分离得到33个化合物,本文鉴定12个化合物的结构,分别为5-羟基-7,4’-二甲氧基黄酮(1)、金合欢素(2)、木犀草素(3)、芫花素(4)、芫花苷(5)、腺苷(6)、芫花素-5-O-β-D-吡喃葡萄糖苷(7)、高次衣草素-7-O-β-D-吡喃葡萄糖苷(8)、次黄嘌呤(9)、尿嘧啶(10)、8-C-β-D-半乳糖基异牡荆素(11)、4-(1,2,3-三羟基丙基)-2,6-二甲氧基苯-1-O-β-D-葡萄糖苷(12)。结论:除化合物1,3,4外,化合物2,5～12均为首次从白木香叶中分离得到。     

The production of nitric oxide and prostaglandin E2 in peritoneal macrophages is inhibited by Andrographis paniculata, Angelica sinensis and Morus alba ethyl acetate fractions

Aim of the study

Traditional Chinese medicine herbs (TCMHs) are used in medicines as well as in daily dietary supplements in Asia. In this study, we employed pNF-κB-Luc or pIFN-γ-Luc and BALB/c mice peritoneal macrophages or splenocytes to investigate both the immune and inflammatory effects of six selected plant species.

Materials and Methods

Specifically, we used ethyl acetate fractions of Astragalus membranaceus (Fisch.) Bunge var. mongholicus (Bunge) Hsiao (Fabaceae) (AM), Andrographis paniculata (Burm. f.) Nees (Acanthaceae) (AP), Angelica sinensis (Oliv.) Diels (Apiaceae) (AS), Eucommia ulmodes Oliv. (Eucommiaceae) leaves (EU leaves), Isatis indigotica Fort. (Brassicaceae) (II) and Morus alba L. (Moraceae) (MA).

Results

We found that ethyl acetate fractions of AP, AS and MA significantly decreased NF-κB luciferase activity and also the secretion of NO and PGE₂ in LPS/IFN-γ stimulated mouse peritoneal macrophages (p < 0.05). In contrast, they did not affect IFN-γ luciferase activity or IFN-γ production in concanavalin A (Con A)-activated mouse splenocytes. Our results indicated that the anti-inflammatory properties of these plant extracts might be resulted from the inhibition of pro-inflammatory mediators (e.g., NO and PGE₂), at least in part via suppression of a signaling pathway such as NF-κB.

Conclusions

Collectively, we have found that three potent bioactive TCMH species exerted significant NF-κB inhibitory activity and acted in a cell type dependent fashion.     

Neuroprotective effect of 1-methoxyoctadecan-1-ol from Uncaria sinensis on glutamate-induced hippocampal neuronal cell death

Ethnopharmacological relevance

We isolated a single compound, 1-methoxyoctadecan-1-ol (MOD), from dried hooks and stems of Uncaria sinensis, which is used in traditional Korean medicine to provide relief from various nervous related symptoms.

Materials and methods

Neuroprotective effects of MOD against glutamate-induced oxidative stress in HT22 cells were investigated by analyzing cell viability, lactate dehydrogenase, flow cytometry, reactive oxygen species (ROS) and Western blot assays.

Results

Exposure to glutamate alone resulted in remarkable hippocampal neuronal cell death; however, pretreatment with MOD resulted in suppression of neuronal death and ROS accumulation in connection with cellular Ca²⁺ level after exposure to glutamate. Stimulation by glutamate also caused significant protein level of phosphorylated p38 mitogen-activated protein kinases (MAPK), and dephosphorylated phosphatidylinositol-3 kinase (PI3K), however, pretreatment with MOD resulted in inhibition of these changes in protein level. Treatment with glutamate alone led to suppressed protein level of mature brain-derived neurotrophic factor (BDNF) and phosphorylated cAMP response element binding protein (CREB); however, pretreatment with MOD resulted in significant enhancement of this level of protein. Anti-oxidant N-acetyl-L-cysteine and both Ca²⁺ inhibitors, BAPTA and EGTA, showed effects similar to those of MOD in all proteins examined, except mature BDNF.

Conclusions

Our results suggest that MOD mainly exerted neuroprotective effects in suppression of ROS accumulation and up-regulation of mature BDNF in association with p38 MAPK and PI3K signaling in hippocampal neuronal cells.     

白木香叶脂溶性化学成分研究

目的:研究白木香叶脂溶性化学成分.方法:应用溶剂法和色谱法分离纯化化合物,利用谱学技术鉴定化合物结构.结果:分离得到11个化合物,分别鉴定为β-谷甾醇(1)、二十六烷酸(2)、隐丹参酮(cryptotanshinone,3)、2α-羟基熊果烷(2α-hydroxyursane,4)、二氢丹参酮I(dihydrotanshinone I,5)、丹参酮I(tanshinone I,6)、丹参酮ⅡA(tanshinoneⅡA,7)、2α-羟基熊果酸(2α-hydroxyursolic acid,8)、对羟基苯甲酸(p-hydroxybenzoic acid,9)、对羟基苯酚(hydroquinone,10)和胡萝卜苷(11).结论:除化合物9外,其他化合物均为首次从白木香叶中分离得到.     

Cordysinocan,a polysaccharide isolated from cultured Cordyceps, activates immune responses in cultured T-lymphocytes and macrophages: Signaling cascade and induction of cytokines

Cordyceps sinensis, a well-known traditional Chinese medicine, possesses activities in anti-tumor, anti-oxidation and stimulating the immune response; however, the identity of active component(s) is not determined. A strain of Cordyceps sinensis, namely UST 2000, has been isolated. By using activity-guided purification, a novel polysaccharide of molecular weight ∼82 kDa was isolated from the conditioned medium of cultured Cordyceps. The isolated exo-polysaccharide, namely cordysinocan, contains glucose, mannose, galactose in a ratio of 2.4:2:1. In cultured T-lymphocytes, application of cordysinocan induced the cell proliferation and the secretion of interleukin-2, interleukin-6 and interleukin-8. In addition, the phosphorylation of extracellular signal-regulated kinases (ERK) was induced transiently by the treatment of cordysinocan. Moreover, application of cordysinocan in cultured macrophages increased the phagocytosis activity and the enzymatic activity of acid phosphatase. These results therefore verify the important role of Cordyceps polysaccharide in triggering such immune responses.     

当归饮片趁鲜切制的可行性探讨

目的:探讨当归饮片趁鲜切制代替传统切制方法的可行性。方法:采用化学手段,建立多指标定量、定性评价的方法,评价不同方法切制的当归饮片质量。结果:趁鲜切制的饮片其主要药效成分藁本内酯的含量均低于传统的二次软化切制方法制备的饮片,2种方法切制饮片的指纹图谱相似度基本在90%以上。结论:以趁鲜切制法代替传统切制方法似乎有些不妥,2种方法切制的饮片临床效果是否有差别,仍需进一步研究。     

白木香树干的化学成分研究

为了研究白木香树干的化学成分,该研究采用硅胶柱色谱与Sephadex LH-20凝胶柱色谱进行分离纯化,并运用波谱方法对所分离的化合物进行结构鉴定。从白木香树干正丁醇萃取物中分离鉴定了16个化合物,经波谱解析鉴定为threo-buddlenol C(1),thero-ficusesquilignan A(2),erythro-buddlenol C(3),(±)-buddlenol D(4),(-)-杜仲树脂酚(5),(-)-松脂素(6),5'-甲氧基落叶松脂醇(7),erythro-guaiacylglycerol-β-coniferyl ether(8), threo-guaiacylglycerol-β-coniferyl ether(9),herpetin(10),(+)-丁香树脂酚(11),curuilignan(12),刺五加酮(13),松伯醇(14),3,4,5-三甲氧基苯酚(15),雪胆甲素(16),其中化合物1~13为木脂素。化合物1~10,14~16均为首次从白木香中分离得到。     

当归多糖对小鼠衰老造血干细胞端粒、端粒酶及P53的影响

目的: 观察当归多糖(ASP)对小鼠造血干细胞(HSC)端粒长度、端粒酶活性及P53表达的影响,探讨ASP调控HSC衰老的可能机制。 方法: C57BL/6J小鼠随机分为正常组、衰老组和干预组,衰老组采用X线全身均匀照射,建立小鼠HSC衰老模型;干预组在照射期间给予ASP灌胃;正常组给予NS灌胃。免疫磁珠分离HSC,运用细胞周期分析和β-半乳糖苷酶(SA-β-Gal)染色观察HSC衰老生物学变化; Western blot检测P53蛋白表达,Southern blot 和TRAP-PCR分别检测HSC端粒长度和端粒酶活性。 结果: 与正常组比较,X线能显著增加衰老组HSC G₁期细胞比例、SA-β-Gal染色阳性细胞率及P53蛋白表达;降低端粒长度和端粒酶活性。与衰老组比较,ASP能显著抑制衰老HSC G₁期细胞比例及SA-β-Gal染色阳性细胞率的增加;下调P53蛋白表达;增加端粒长度和端粒酶活性。 结论: ASP能够拮抗X线诱导的HSC衰老,其作用机制可能与增加端粒长度及端粒酶活性、下调P53蛋白表达有关。     

白木香乙酰乙酰基辅酶A硫解酶基因(AsAACT)的克隆与表达分析

目的：对国产沉香的基原植物白木香Aquilaria sinensis （Lour.）Gilg乙酰辅酶A酰基转移酶（acetyl-CoA C-acetyl transferase,AACT）基因AsAACT全长进行克隆并展开生物信息学分析和表达分析,为解析沉香萜类次生代谢产物的生物合成机制奠定基础。方法：根据获得的白木香转录组数据库AACT部分转录本序列设计引物,采用RT-PCR及RACE技术,以白木香茎cDNA为模板,克隆获得AsAACT全长,进行生物信息学分析;采用荧光定量PCR,以GADPH为内参,分析白木香愈伤组织受不同伤害胁迫AsAACT的表达模式。结果：AsAACT开放阅读框（opening reading frame,ORF）为1 236 bp,编码411个氨基酸残基,酶命名为AsAACT;白木香愈伤受物理伤害（切割）后表达量没有明显变化,但受化学伤害（MeJA）后4 h表达量升高了5.5倍,说明该基因对MeJA诱导的化学伤害较敏感,且能够在早期响应伤害胁迫。结论：通过AsAACT基因的全长cDNA克隆和表达特性分析,为后续深入研究其在沉香倍半萜合成途径的功能奠定基础。     

土鳖虫纤溶酶编码区序列的克隆与表达

目的:获得土鳖虫纤溶酶编码区序列,并进行原核和真核表达。方法:根据已报道的多种动物纤溶酶基因cDNA序列设计引物,用RT-PCR和3′RACE法克隆得到土鳖虫纤溶酶编码区序列;将该序列克隆进入大肠杆菌和毕赤酵母进行表达。结果:序列分析表明,所克隆的纤溶酶编码区序列长672bp,共编码224个氨基酸残基,起始氨基酸序列为IVGG,与多种动物纤溶酶一致。将此cDNA序列在大肠杆菌和毕赤酵母中进行表达,前者获得没有活性的表达蛋白,后者获得具有纤溶活性的重组表达蛋白。结论:首次报道了土鳖虫纤溶酶编码区序列,并进行了初步表达,为进一步研究其功能奠定了基础。     

Study to establish the role of JAK2 and SMAD1/5/8 pathways in the inhibition of hepcidin by polysaccharides from Angelica sinensis

Ethnopharmacological relevance

Angelica sinensis polysaccharide (ASP) is one of the major active ingredients in Angelica sinensis (Oliv.) Diels. This traditional Chinese medicine has been used for thousands of years for treating gynecological diseases.

Aim of the study

Previous studies have suggested that ASP from the roots of Angelica sinensis (Oliv.) Diels suppresses hepcidin expression, but the underlying molecular mechanisms are not known. The present study was designed to establish the role of the janus-kinases 2 (JAK2) and son of mothers against decapentaplegic 1/5/8 (SMAD1/5/8) pathways in the inhibition of hepcidin by polysaccharides from Angelica sinensis in normal rats.

Materials and methods

ASP was administered orally (0.3, 0.6 and 1.2 g/kg body weight) to male Sprague–Dawley rats every day for 20 days. Intraperitoneal injections of recombinant human erythropoietin (rhEPO; 800 and 2000 U/kg body weight) were given to the positive control group every day for 3 days. After administration, hepcidin levels, blood parameters, serum iron status and non-heme iron concentrations in the liver were examined. Western blot analyses were used to investigate the expression of five relevant signaling proteins in the liver.

Results

RhEPO injection significantly stimulated erythropoiesis and expression of the serum transferrin receptor (sTfR), and decreased serum iron status and non-heme iron concentrations in the liver. However, blood parameters barely changed in the ASP groups. sTfR, serum iron, and liver iron levels altered only in the ASP high-dose group (1.2 g/kg body weight). rhEPO and ASP significantly reduced hepcidin expression by inhibiting the expression of phospho-SMAD1/5/8 and JAK2 in the liver, but not through transmembrane protease serine 6 (TMPRSS6) and extracellular signal-regulated kinase 1/2 (ERK1/2).

Conclusions

These data suggested that ASP can interrupt the JAK2 and SMAD1/5/8 pathways, which eventually results in lower expression of hepcidin.     

白木香内生真菌多节孢 Nodulisporium sp. A4的化学成分研究

目的:研究来源于药用植物白木香、具有抗肿瘤活性的内生真菌多节孢Nodulisporium sp.A4的化学成分.方法:采用马铃薯葡萄糖(PD)液体培养基培养、乙酸乙酯萃取得到内生真菌A4的发酵液和菌丝体粗提物,采用正相硅胶柱色谱、反相硅胶柱色谱、葡聚糖凝胶Sephadex LH-20柱色谱、制备薄层色谱和重结晶等方法对粗提物进行分离纯化,运用现代波谱技术(NMR,MS)结合文献对照鉴定化合物结构.采用MTT法测定化合物的体外抗肿瘤活性.结果:从内生真菌A4的发酵液中分离获得7个化合物,分别为5-methyl-2-vinyltetrahydrofuran-3-ol(1),6-methyl-2-(5-methyl-5 -vinyltetrahydrofuran-2 -yl)hept-5-en-2-ol (2),6α-hydroxycyclonerolidol(3),rel-( 1S,4S,5R,7R,10R) -10-desmethyl-1 -methyl-11 -eudesmene(4),酪醇(5),8-甲氧基-1-萘酚(6),1,8-二甲氧基萘(7).从内生真菌A4的菌丝体中分离获得3个化合物,分别为麦角甾醇(8),过氧化麦角甾醇(9),啤酒甾醇(10).药理活性评价结果表明在作用浓度为100 mg·L-1时,化合物3和4对肿瘤细胞SF-268和NCI-H460的抑制率分别为89.1％,44.2％和82.3％,79.8％.结论:化合物1为新的天然产物,化合物2,3,7,10为首次从该属真菌中分离得到,化合物3和4对肿瘤细胞SF-268和NCI-H460的增殖具有一定的抑制作用.     

不同加工方法对当归多糖的影响

目的:开展不同加工方法对当归多糖含量影响的研究,以期为建立当归药材合理的加工工艺提供依据.方法:以甘肃岷县同一采收期的当归为研究对象,应用不同加工方法制备样品,采用苯酚-硫酸法及硫酸-咔唑法显色,紫外分光光度法测定不同加工方法当归样品的中性、酸性多糖含量;MTT法开展2种外观性状及含量差异较大的不同加工方法制备的当归多糖对小鼠脾淋巴细胞增值影响的研究.结果:紫外分光光度法测得微波中火干燥法所得当归多糖含量最高(26.0%),真空0.09MPa,50℃干燥法所得当归多糖含量最低(2.25%);微波干燥法制备的当归多糖(P<0.01)在高浓度组既能直接促进小鼠脾脏淋巴细胞增殖,又能增加伴刀豆球蛋白A诱导的T淋巴细胞增殖,而远红外干燥法制备的当归多糖在任何浓度及条件下均无明显活性.结论:不同加工方法对当归多糖含量及小鼠脾淋巴细胞增值均会造成显著影响.     

香椿子总多酚对心肌缺血再灌注大鼠的保护作用

目的:观察香椿子总多酚对心肌缺血再灌注大鼠的保护作用。方法:50只SD大鼠随机分成假手术组:冠脉下穿线不结扎+0.5%羧甲基纤维素钠、模型组:冠脉下穿线结扎+0.5%羧甲基纤维素钠、香椿子总多酚低剂量(XD,50 mg·kg-1)+冠脉下穿线结扎、中剂量(XZ,100 mg·kg-1)+冠脉下穿线结扎、高剂量(XG,200 mg·kg-1)+冠脉下穿线结扎组,各组均ig给药。采用左冠状动脉前降支结扎30 min再灌120 min的方法复制大鼠心肌缺血再灌注损伤模型。再灌注结束时取血清,测定血清中磷酸肌酸激酶(CK)、肌钙蛋白I(cTnI)、超氧化物歧化酶(SOD)、丙二醛(MDA)的水平。取出心脏,自结扎线以下平行均匀切成5片,用1%氯化三苯基四氮唑(TTC)浸染15 min,计算心肌梗死程度。结果:与模型组相比,香椿子总多酚处理组能有效降低心肌缺血再灌注大鼠血清中CK,cTnI,MDA的水平,增加SOD活力,减轻心肌梗死程度。结论:香椿子总多酚对心肌缺血再灌注大鼠具有一定的保护作用。