湿地匍灯藓(Plagiomnium acutum)的防御机制对镉胁迫的响应

采用浸没培养实验研究不同浓度镉溶液(0、2、5、50mg·L-1)对湿地匍灯藓(Plagiomnium acutum)的膜系统、光合系统的损伤情况及其抗氧化酶系统的影响。结果表明:湿地匍灯藓外表伤害症状随Cd胁迫浓度增加而加重;湿地匍灯藓对Cd有较强的生物富集能力,且藓体Cd累积量与溶液Cd浓度显著正相关(P<0.05);湿地匍灯藓的总叶绿素含量随Cd浓度增加显著降低,其含量与可溶性蛋白含量显著正相关(P<0.05);Cd胁迫引起湿地匍灯藓的丙二醛(MDA)积累,同时伴随可溶性糖和游离脯氨酸含量显著增加,其中可溶性糖含量与MDA含量显著正相关(P<0.05);可溶性蛋白含量和过氧化氢酶(CAT)活性随Cd浓度增加均持续下降,而超氧化物歧化酶(SOD)和过氧化物酶(POD)的活性却随Cd浓度增加而呈现逐渐上升趋势,其中可溶性蛋白和CAT活性均与MDA含量显著负相关(P<0.05),SOD活性则与MDA含量显著正相关(P<0.05),POD活性与MDA含量极显著正相关(P<0.01)。Cd胁迫抑制或降解光合作用相关酶类、细胞膜质过氧化和蛋白质代谢紊乱可能是湿地匍灯藓遭受Cd毒害的主要原因;湿地匍灯藓通过增加可溶性糖含量、SOD和POD活性来抵御Cd的毒害,但由于CAT活性丧失,使湿地匍灯藓的防御系统崩溃,最终导致了湿地匍灯藓细胞的凋亡。可溶性糖、SOD和POD可以作为湿地匍灯藓防御Cd毒害的指标。

Cd、Pb复合胁迫下湿地匍灯藓(Plagiomnium acutum)重金属累积效应及抗氧化系统响应

采用浸没培养方法研究了不同浓度Cd、Pb复合胁迫下湿地匍灯藓(Plagiomnium acutum)重金属的累积效应及抗氧化系统的响应。结果显示:复合胁迫下Pb显著抑制湿地匍灯藓对Cd的吸收,与低Pb水平(20 mg·L-1)相比(对照无检出值),在低Cd(2mg·L-1)处理时,随Pb处理浓度增加,藓体内Cd累积量分别降低了30.1%和78.4%;在高Cd(50 mg·L-1)水平处理时,其降幅则分别达到了35.0%和86.9%。复合胁迫引起湿地匍灯藓脂质过氧化终产物丙二醛(MDA)含量随胁迫程度加大而持续升高,当复合胁迫浓度达到最大时,MDA含量也达到了最大,与对照相比显著增加7162%。与此同时,超氧化物歧化酶(SOD)活性和脯氨酸含量也呈现了应激性增加,且均在最高复合胁迫浓度处理时达到最大,与对照相比分别增加237.4%和2049%,而过氧化氢酶(CAT)和过氧化物酶(POD)活性则持续降低。基于此,可以认为:Cd、Pb复合胁迫下,Pb对Cd的累积效应有明显的抑制作用,且这种抑制作用有浓度依赖性;Cd、Pb复合胁迫所致细胞氧化损伤的主要来源为过氧化物的清除能力低下所致过氧化物积累。

Cd、Pb胁迫下湿地匍灯藓(Plagiomnium acutum)叶绿素荧光特性研究

采用浸没培养实验研究了不同浓度Cd(0、1、5、10 mg·L-1)、Pb(0、10、25、50 mg·L-1)单一以及二者低、中、高浓度组合复合处理对湿地匍灯藓(Plagiomnium acutum)叶绿素荧光以及细胞膜透性的影响。结果表明:(1)湿地匍灯藓Fv/Fm、Fv/Fo、Fv′/Fm′、ETR、Y(II)的下降程度与Cd、Pb胁迫浓度及胁迫时间存在明显的剂量、时间-效应关系;随胁迫时间延长,Y(NO)总体呈现持续上升的趋势,qN表现为先升后降,而qP则呈现出先降后升趋势。(2)湿地匍灯藓的叶绿素荧光参数对Pb、Cd胁迫的响应存在差异,单一Pb胁迫初期各荧光参数即出现不同程度的下降,单一Cd胁迫下上述参数均在处理第4 d才出现明显下降;在胁迫第1 d,中、低浓度Cd、Pb单一及复合胁迫均引起湿地匍灯藓的Fv/Fm、Fv/Fo、Fv′/Fm′增加。(3)Cd、Pb胁迫均使处理液电导率升高,升幅大小顺序为Cd/Pb复合胁迫>Cd胁迫>Pb胁迫。表明湿地匍灯藓在Cd、Pb胁迫初期可通过应激性增加PSII活性中心数来抵御重金属的毒害,但由于电子传递和热耗散能力较弱,使PSII活性中心受损,最终导致光合系统损伤;Cd、Pb复合胁迫对湿地匍灯藓细胞膜的损伤和毒害作用显著强于单一胁迫,Cd对细胞膜的毒害作用又强于Pb;Fv/Fo可作为Cd、Pb胁迫的敏感指标。

湿地匍灯藓(Plagiomnium acutum)对Pb胁迫的生物标志物响应

采用浸没培养实验研究了不同浓度Pb胁迫下湿地匍灯藓(Plagiomniumacutum)的总叶绿素、丙二醛、可溶性糖、游离脯氨酸含量以及抗氧化酶活性等生理生化生物标志物的变化,以探讨Pb胁迫下湿地匍灯藓的受损和耐受机理。结果表明,湿地匍灯藓外表伤害症状与Pb胁迫浓度存在明显的剂量-效应关系;低浓度(20mg·L-1)Pb胁迫可以显著地促进湿地匍灯藓的总叶绿素含量增加,而中、高浓度(50mg·L-1)Pb胁迫则导致总叶绿素含量显著降低。湿地匍灯藓对Pb具有较强的生物富集能力,且藓体Pb的累积量与环境Pb浓度呈显著正相关。湿地匍灯藓体内MDA和游离脯氨酸含量随Pb胁迫浓度的增加表现为显著升高。可溶性糖含量仅在高浓度Pb胁迫下显著升高。可溶性蛋白含量、过氧化物酶(POD)和过氧化氢酶(CAT)活性均随Pb胁迫浓度的增加而显著下降,其中CAT活性的降幅最大,SOD活性随Pb胁迫浓度的增加逐渐显著增加。在高浓度Pb胁迫下,由于细胞膜脂过氧化和蛋白质变性失活所导致的膜保护体系损伤可能是湿地匍灯藓Pb中毒的主要原因,而渗透调节可能是其缓解Pb毒害的一种主要方式,SOD则在清除Pb胁迫产生的活性氧自由基过程中起重要作用。总叶绿素、丙二醛、游离脯氨酸、可溶性蛋白、CAT和SOD可以作为湿地匍灯藓受Pb胁迫的敏感生物标志物。

A New Alkaloid from Sinomenium acutum

Sinomenium acutum is widely used in East and South Asia for the treatment of many diseases, especially rheumatoid arthritis （RA）. The chemical research on Sinomenium acutum led to the isolation of a new alkaloid compound （1）. On the basis of chemical evidences and spectral analysis, 1 was identified as N-（1, 7-dimethoxylphenanthren-2-yl）acetamide.

Mathematical Models for Some Ecophysiological Characteristics in Different Weather Condition in Moss (Plagiomnium acutum)

Based on the data from experiment, several mathematical models for describing the ecophysiological characteristics ofPlagiomnium acutum and the relationship between photosynthetic active radiation (R)P, temperature of atmospheric (T a), relative humidity (H r), concentration of carbon dioxide were established with the method of regression. The biological meanings of the models were analyzed primarily, which showed significance in both theory and application.

Systematic Review and Meta-Analysis on the Effect of Transdermal Preparations of Sinomenium Acutum on Rheumatoid Arthritis

Objective:We evaluated the efficacy and safety of transdermal preparations of Sinomenium acutum(SA)for rheumatoid arthritis(RA).Methods:Randomized controlled trials(RCTs)of SA transdermal preparations for RA were extracted from relevant databases and screened in accordance with the inclusion criteria.The Cochrane System Evaluation Manual(version 5.1.0)was used to assess the quality of the included trials.We used the Cochrane Review Manager(version 5.4)to conduct the meta-analysis.Results:Six trials comprising 436 patients(220 patients in the treatment group and 216 patients in the control group)were analyzed.The meta-analysis indicated that SA transdermal preparations in combination with disease-modifying antirheumatic drugs(DMARDs)enhanced the overall effect(odds ratio[OR]3.97,95%confidence interval[CI][2.25,7.00],P<0.00001),decreased visual analogue scale(VAS)results(mean difference[MD]-0.64,95%CI[-1.20,-0.09],P=0.02),decreased laboratory indexes including the erythrocyte sedimentation rate(ESR)(MD-4.36,95%CI[-5.63,-3.08],P<0.00001)and C-reactive protein(CRP)(MD-3.6,95%CI[-3.99,-3.21,P<0.00001]),and decreased the Disease Activity Score-28(DAS28)(MD-0.41,95%CI[-0.78,-0.03],P=0.03).The results suggest that combination therapy did not shorten the duration of morning stiffness(DMS;standardized MD[SMD]-6.13,95%CI[-17.33,5.06],P=0.28)or reduce rheumatoid factor(RF)laboratory indexes(SMD-0.85;95%CI[-2.19,0.49],P=0.21).Only one study reported adverse reactions,and thus,it was difficult to determine whether adverse drug reactions in the combination therapy group were significantly different from those in the control group.Conclusion:We found that SA transdermal preparations combined with DMARDs may have greater clinical efficacy than DMARDs for RA.More well-designed and high-quality RCTs are required to verify the findings and determine whether transder-mal preparations cause fewer adverse events.

Chloroplast DNA reveals genetic population structure in Sinomenium acutum in subtropical China

Objective:The population density and diversity of Sinomenium acutum(Menispermaceae)have been greatly reduced recently by overharvesting for medicinal purposes in China.Therefore,it is urgent that the remaining populations are investigated,and that strategies for the utilization and conservation of this species are developed.This study aimed to find the possible glacial refugia and define the genetic diversity of S.acutum for its proper utilization and conservation.Methods:A total of 77 S.acutum samples were collected from four locations,Qinling Mountains,Daba Mountains,Dalou Mountains,and Xuefeng Mountains,in subtropical China.Genetic diversity among and between these populations were phylogenetically analyzed using four chloroplast DNA molecular markers(atpI-atpH,trnQ-50 rps16,trnH-psbA and trnL-trnF).Results:A total of 14 haplotypes(C1 to C14)were found in collected samples.Haplotypes C1 and C3 were shared among all populations,with C3 as the ancestral haplotype.Haplotypes C11 and C12 diverged the most from C3 and other haplotypes.No obvious phylogeographic structure was found in four locations using the GST/NST test.There is no evidence of rapid demographic expansion in S.acutum based on the mismatch distribution,and the results of Tajima’s D test,and Fu’s FS test.Our analyses of molecular variance revealed a high level of genetic variation within populations.In contrast,the genetic differentiation among S.acutum populations was low,indicating frequent gene flow.Conclusion:Xuefeng,Dalou,and Daba Mountains were possible glacial refugia for the populations of S.acutum.C1,C3,C11 and C12 haplotypes of S.acutum should be carefully preserved and managed for their genetic value.

青藤Sinomenium acutum(Thunb.)Rehd.et Wils全基因组调查

青藤是一种重要的中药材,具有较高的药用价值。目前,对于青藤的分子生物学研究极为有限,基因资源比较匮乏。全基因组的破译将为其分子生物学研究的深入开展提供坚实的基础,而对青藤进行全基因组调查以明确其基因组特征可以为全基因测序方案的制定提供依据。本研究基于Illumina测序平台测定了青藤基因组,结合生物信息学方法得到基因组大小、杂合率、重复序列等特征,并采用SOAPdenovo软件对基因组进行初步组装。过滤得到168.72 G有效数据,青藤基因组大小估计为829.72 Mb;重复序列比率为51.72%,杂合率为0.48%,GC含量为34.57%;K-mer=35进行初步组装得到contigs N50为162 bp,总长为621 033 264 bp,scaffolds N50为489 bp,总长为780 536 487 bp。鉴于青藤基因组高重复序列的特点,其全基因测序可采用Illumina测序技术和Pac Bio测序技术相结合的策略,并采用Hi-C技术以及其相应的组装软件,以此获得青藤全基因组图谱。

Bioinformatics Based Therapeutic Effects of Sinomenium Acutum

Objective: To decipher the possible mechanisms of Sinomenium Acutum(SA) in treating diseases by a bioinformatics method. Methods: SA ingredients were searched according to Chinese Pharmacopoeia, Chinese Medicine Dictionary and Traditional Chinese Medicines Database(TCMD). Active compounds and target proteins of SA were acquired through the Pubchem platform. Pathway, network and function analyses of SA were performed with ingenuity pathway analysis(IPA), a bioinformatics analysis platform. Disease, biofunction-target networks were established with Cytoscape. Results: Eighteen ingredients from SA were obtained. Seven active ingredients with 31 active target proteins were acquired according to PubChem Bioassay test. By IPA analysis, 277 canonical pathways belonging to 17 function categories were collected, 23 kinds of diseases, 21 categories bio-functions were obtained. Based on P value, calculated by IPA, the top 5 significant pathway of SA targets include phosphatidylinositol 3 kinase/Akt(PI3 K/Akt) signaling, prostate cancer signaling, macrophage migration inhibitory factor(MIF) regulation of innate immunity, Guanosine-binding protein coupled receptor(GPCR) signaling, and ataxia telangiectasia mutated protein(ATM) signaling. Disease and bio-function network analysis indicated that mitogen activated protein kinase 1(MAPK1), MAPK3, p65 nuclear factor κB(RELA), nuclear factor of κB inhibitor alpha(NFκBIA), interleukin 1β(IL-1β), prostaglandin G/H synthase 2(PTGS2) and tumor protein 53(TP53) were the critical targets in various diseases treated by SA. Conclusions: In the different view of target, pathway, disease and bio-function, inflammation was found to be a central theme in many chronic conditions. SA could be used not only as an anti-inflammatory agent, but also for the treatment of cancers, neurological diseases, psychological disorders and metabolic diseases.

烟头引燃藏柏林下尖叶匐灯藓层研究

为研究地表苔藓层的引燃特性,以昆明地区广泛分布的藏柏林下地被物尖叶匐灯藓为研究对象,在室内模拟构造野外自然生长状态的尖叶匐灯藓床层,设定5个水平含水率(0%、5%、10%、15%、20%)以及6个水平风速(0、1、2、3、4、5 m·s^(-1))的烟头引燃藏柏林下尖叶匐灯藓层试验。结果表明:烟头引燃尖叶匐灯藓层的最大含水率约为20%;含水率和风速以及两个因子之间的相互作用对尖叶匐灯藓层的引燃比例有显著影响。当风速≤2 m·s^(-1)时,尖叶匐灯藓层不能被烟头引燃;当风速从3 m·s^(-1)增加到5 m·s^(-1)时,含水率从20%降至0%时,烟头的引燃比例从13%增至82%。建立了烟头引燃尖叶匐灯藓层的二元Logistic回归模型,预测的准确率达71.7%。研究结果可为昆明地区地表可燃物的管理、火险预警等提供依据,为深入开展苔藓植物的火灾风险辨识与评价奠定基础。

高温胁迫下两种藓类植物过氧化物酶活性的变化

在不同的高温胁迫条件下 ,对湿地匍灯藓 (Plagiomnium acutum)和大羽藓 (Thuidium cymbifolium)过氧化物酶 (POD)活性及其与处理时间和处理温度的关系进行了初步研究。结果表明 ,在一定的温度范围内 ,随着温度的升高 ,POD活性增加 ,二者成线形关系。在一定温度条件下 ,一般随着处理时间的延长 ,POD活性增加。但是当超过一定的温度 (4 5～ 5 0°C)以及一定的处理时间 (4～ 6h) ,POD活性有所下降。结果还表明 ,湿地匍灯藓的POD活性显著高于大羽藓。而且在高温胁迫下 ,湿地匍灯藓 POD活性变化比大羽藓活跃 ,其变化幅度也比大羽藓大。

汉防己多糖对急性酒精性肝损伤小鼠氧化应激及肝细胞凋亡的影响

目的:探讨汉防己多糖对急性酒精性肝损伤小鼠的保护作用及其机制。方法:将60只小鼠随机分为空白对照组(生理盐水)、模型组(生理盐水)、联苯双酯组(阳性对照,150 mg/kg)和汉防己多糖低、中、高剂量组(100、200、400 mg/kg),每组10只,ig给药,每天1次,连续7 d。末次给药1 h后,除空白对照组外其余各组小鼠均ig 50%乙醇(0.1 m L/10 g)溶液复制急性酒精性肝损伤模型。12 h后,测定小鼠血清中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)水平以及肝组织中丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽(GSH)、谷胱甘肽过氧化物酶(GSH-Px)水平;苏木精-伊红染色观察小鼠肝组织病理改变;流式细胞术检测肝细胞凋亡率。结果:与空白对照组比较,模型组小鼠肝组织发生水肿、细胞排列紊乱及局部坏死等病变,血清中ALT、AST水平和肝组织中MDA水平以及肝细胞凋亡率显著升高,肝组织中SOD、GSH、GSH-Px水平显著降低,差异均有统计学意义(P<0.01)。与模型组比较,汉防己多糖中、高剂量组小鼠肝组织细胞变性和坏死程度均减轻;除汉防己多糖低剂量组小鼠肝细胞凋亡率降低不明显外,其余各给药组小鼠上述指标均显著改善(P<0.05或P<0.01)。结论:汉防己多糖对急性酒精性肝损伤小鼠具有明显的保护作用,其机制可能与抗氧化应激及减少肝细胞凋亡有关。

青风藤的研究进展

从生药学、化学成分、药理作用及临床应用等方面综述了青风藤的研究进展,并对进一步研究和开发青风藤的目标提出了展望。

毛细管电泳高频电导法测定青风藤药材及制剂中青藤碱的含量

A method based on capillary electrophoresis with a high frequency conductivity detection was developed for the determination of Sinomenine. The key factors for separation and determination were studied and the best analysis conditions were obtained. In the buffer of 2.0 mmol/L HAc-3.0 mmol/L NaAc-10.0% CH 3OH at a separation voltage of 22 kV, Sinomenine in the sample could be separated and detected within 4 min. The linear mass concentration of Sinomenine is in the range of 1.0—36.0 μg/mL, the limit of detection reached 0.20 μg/mL. The method was used for the analysis of Sinomenine in Sinomenium acutum and tablets satisfactorily with a recovery of 96%—102%.

脱水与复水过程中湿地匍灯藓的生理生化响应

研究了脱水与复水过程中湿地匍灯藓的渗透调节能力、抗氧化保护系统以及DNA损伤与修复的影响。结果表明:(1)在脱水过程中,游离脯氨酸、可溶性糖、还原性糖含量均出现明显增加,脱水12h时达到最大;细胞膜透性上升;超氧化物歧化酶、过氧化氢酶和过氧化物酶活性均持续上升,抗坏血酸含量则逐渐降低;DNA损伤明显加大,脱水24h时DNA已全部降解为低分子量片段。(2)在复水过程中,以上各项指标变化趋势与脱水处理时刚好相反。表明湿地匍灯藓具备复苏植物的典型特征,在含水量变化(变水)过程中具有较强的抗干(旱)性,其抗逆性主要体现为修复能力。

反相离子对色谱法测定青风藤中青藤碱的含量

目的建立离子对色谱法测定青风藤中青藤碱的含量。方法使用Eclipse Plus C18柱（4.6 mm×250 mm,5μm）;乙腈：磷酸盐缓冲液（0.03 mol/L磷酸二氢钾和0.03 mol/L庚烷磺酸钠,磷酸调节pH值至4.0）=18：82为流动相,流速1.0 ml/min,柱温40℃,检测波长262 nm。结果青藤碱在0.025-0.5 mg/ml（r=1.000）范围内呈良好的线性关系。最低检测限为2.5μg/ml,低、中、高3个浓度的日内精密度RSD分别为0.62%、0.35%、0.60%,日间精密度RSD为1.54%、0.70%、0.45%。平均回收率99.9%（RSD=2.9%,n=6）,均符合分析要求。结论该方法简单快捷,结果准确,适用于测定青风藤中青藤碱的含量。

青藤与毛青藤茎中青藤碱积累的组织化学研究

青藤及毛青藤茎中青藤碱积累的组织化学研究表明，青藤碱主要分布在皮层和接近维管形成层的射线细胞中。在髓鞘及细胞壁已木质化加厚的射线细胞内也有少量青藤碱分布，且木化的细胞壁中浸透着青藤碱。在髓鞘及射线细胞中有大量的淀粉粒。根据不同直径茎的测试，细胞中淀粉粒的含量随茎的直径增粗而逐渐增加，而细胞中青藤碱的含量在直径小于０．５ｃｍ的茎中较少，以后逐渐增加，到直径为１ｃｍ时青藤碱的含量趋于稳定。因此，直径１ｃｍ以上的藤茎为适宜的采收部分。在两种植物中。

青风藤中总生物碱的气相色谱-质谱联用分析

目的采用索氏提取法对青风藤中总生物碱进行提取分离,并利用气相-质谱联用(GC-MS)方法,对其化学成分进行分析。方法采用90%无水乙醇提取,盐酸酸化后,氨水碱化后三氯甲烷萃取,利用GC-MS对总生物碱中的化学成分进行分离分析,并对其中的15种化学成分进行鉴定。结果该方法提取时青藤碱含量高,达到80.18%,安贝灵碱3.44%,光千金藤碱3.05%。采用峰面积归一化法计算各成分相对百分含量,所鉴定的成分占总流出峰面积的95.1%。结论实验结果为进一步开发研究青风藤中生物碱提供依据。

青风藤化学成分的分离与鉴定

目的对青风藤（Sinomenium auctum（Thunb．）Rehd．et Wils．）化学成分进行分离、鉴定。方法适量干燥青藤茎用10倍量体积分数为70％的乙醇水溶液提取，减压回收乙醇，所得干浸膏研成细粉后，分别用环己烷、氯仿、甲醇超声溶取；采用ODS中低压柱色谱、制备型HPLC方法进行分离，并通过^1H-NMR、^13C-NMR、ES-MS等谱学技术对化合物进行结构鉴定。结果分离得到5个化合物，分别鉴定为穆坪马兜铃酰胺（N-trans-feruloyl tyramine, 1）、syringaresinol mono-β-D-glucoside（2）、1，2，4／3，5-cyclohexanepentol（3）、3-methoxy-6-hydroxy-17-methylrnorphinane（4）、青藤碱（sinomenine ，5）。结论化合物1～4为首次从防己属中分离得到。