Protective mechanism of Paeoniae Radix Alba against chemical liver injury based on network pharmacology,molecular docking,and in vitro experiments

Objective:To explore and validate the potential targets of Paeoniae Radix Alba(P.Radix,Bai Shao)in protecting against chemical liver injury through network pharmacology,molecular docking technology,and in vitro cell experiments.Methods:Network pharmacology was used to identify the common potential targets of P.Radix and chemical liver injury.Molecular docking was used to fit the components,which were subsequently verified in vitro.A cell model of hepatic fibrosis was established by activating hepatic stellate cell(HSC)-LX2 cells with 10 ng/mL transforming growth factor-β1.The cells were exposed to different concentrations of total glucosides of paeony(TGP),the active substance of P.Radix,and then evaluated using the cell counting kit-8 assay,enzyme-linked immunosorbent assay,and western blot.Results:Analysis through network pharmacology revealed 13 key compounds of P.Radix,and the potential targets for preventing chemical liver injury were IL-6,AKT serine/threonine kinase 1,jun protooncogene,heat shock protein 90 alpha family class A member 1(HSP90AA1),peroxisome proliferator activated receptor gamma(PPARG),PTGS2,and CASP3.Gene Ontology(GO)enrichment analysis indicated the involvement of response to drugs,membrane rafts,and peptide binding.Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis revealed that the main pathways involved lipid and atherosclerosis and chemical carcinogenesis-receptor activation.Paeoniflorin and albiflorin exhibited strong affinity for HSP90AA1,PTGS2,PPARG,and CASP3.Different concentrations of TGP can inhibit the expression of COL-I,COL-III,IL-6,TNF-a,IL-1β,HSP-90a,and PTGS2 while increasing the expression of PPAR-γand CASP3 in activated HSC-LX2 cells.Conclusion:P.Radix primarily can regulate targets such as HSP90AA1,PTGS2,PPARG,CASP3.TGP,the main active compound of P.Radix,protects against chemical liver injury by reducing the inflammatory response,activating apoptotic proteins,and promoting the apoptosis of activated HSCs.

Radix Paeoniae Alba attenuates Radix Bupleuri-induced hepatotoxicity by modulating gut microbiota to alleviate the inhibition of saikosaponins on glutathione synthetase

Radix Bupleuri(RB)is commonly used to treat depression,but it can also lead to hepatotoxicity after longterm use.In many anti-depression prescriptions,RB is often used in combination with Radix Paeoniae Alba(RPA)as an herb pair.However,whether RPA can alleviate RB-induced hepatotoxicity remain unclear.In this work,the results confirmed that RB had a dose-dependent antidepressant effect,but the optimal antidepressant dose caused hepatotoxicity.Notably,RPA effectively reversed RB-induced hepatotoxicity.Afterward,the mechanism of RB-induced hepatotoxicity was confirmed.The results showed that saikosaponin A and saikosaponin D could inhibit GSH synthase(GSS)activity in the liver,and further cause liver injury through oxidative stress and nuclear factor kappa B(NF-kB)/NOD-like receptor thermal protein domain associated protein 3(NLRP3)pathway.Furthermore,the mechanisms by which RPA attenuates RBinduced hepatotoxicity were investigated.The results demonstrated that RPA increased the abundance of intestinal bacteria with glycosidase activity,thereby promoting the conversion of saikosaponins to saikogenins in vivo.Different from saikosaponin A and saikosaponin D,which are directly combined with GSS as an inhibitor,their deglycosylation conversion products saikogenin F and saikogenin G exhibited no GSS binding activity.Based on this,RPA can alleviate the inhibitory effect of saikosaponins on GSS activity to reshape the liver redox balance and further reverse the RB-induced liver inflammatory response by the NFkB/NLRP3 pathway.In conclusion,the present study suggests that promoting the conversion of saikosaponins by modulating gut microbiota to attenuate the inhibition of GSS is the potential mechanism by which RPA prevents RB-induced hepatotoxicity.

白芍(Radix paeoniae alba)提取物对中华绒螯蟹(Eriocheir sinensis)造血组织发育及造血因子Astakine的影响

血淋巴细胞在甲壳动物免疫反应中起着非常重要的作用,其主要由造血组织(Hematopoietic tissue,HPT)生成并随着开管式循环分布到身体各处。到目前为止,还没有相关研究表明甲壳动物造血组织是否受草本植物的影响。本研究以中华绒螯蟹(Eriocheir sinensis)为对象,利用荧光定量PCR、免疫荧光和组织化学等技术,初步探索了白芍(Radix paeoniae alba)提取物对中华绒螯蟹造血组织和造血因子Astakine的影响。结果显示,在原代培养的造血组织细胞中添加终浓度为0.85 mg/mL的白芍提取物后,造血因子Astakine mRNA的表达量显著上调(P<0.05)。进一步制备含1%白芍提取物的中华绒螯蟹饲料并进行饲喂实验,结果显示饲喂含白芍提取物饲料的中华绒螯蟹造血组织的Astakine mRNA的表达量较饲喂前上调了10.5倍,要高于对照组。免疫荧光结果同样表明,饲喂含白芍提取物饲料后中华绒螯蟹造血组织的Astakine蛋白荧光信号也要强于对照组。通过造血组织的形态学观察和造血组织体重百分比的检测,我们发现白芍组中华绒螯蟹造血组织含有较多卵形小叶,且造血组织体重百分比较饲喂前增加了6.64%,而对照组仅增加1.6%。免疫保护率实验显示,在脂多糖急性刺激下,白芍添加组的中华绒螯蟹的存活率(77.5%)要高于对照组(57.5%)。综上所述,白芍提取物可以促进中华绒螯蟹造血因子的表达和造血组织的发育,从而提高中华绒螯蟹的免疫调节功能。

24个白桑(Morus alba L．)地方品种的遗传多样性分析

采用ISSR分子标记技术对山东、河北地区的24个白桑（Morus alba L．）地方品种资源进行了遗传多态性分析。筛选的13条ISSR引物共扩增86条扩增带，其中多态性条带63条，多态性比率为73．25％，ISSR标记遗传相似系数范围在0．6706～0．9529。通过类平均聚类（UPGMA）法分析，24份材料聚分为2大类。

Mosquito larvicidal and ovicidal properties of Eclipta alba(L.) Hassk(Asteraceae) against chikungunya vector,Aedes aegypti(Linn.)(Diptera:Culicidae)

Objective:The present study deals with the investigation of larvicidal and ovicidal activities of benzene,hexane,ethyl acetate,methanol and chloroform leaf extract of Eclipta alba（E.alba） against dengue vector,Aedes aegypti（Ae.Aegypti）.Methods:Twenty five earlyⅢinstar larvae of Ae.aegypti was exposed to various concentrations（50-300 ppm） and was assayed in the laboratory by using the protocol of WHO 2005;the 24 h LC50 values of the E.alba leaf extract was determined by Probit analysis.For ovicidal activity,slightly modified method of Su and Mulla was performed.The ovicidal activity was determined against Ae.aegypti to various concentrations ranging from 100-350 ppm under the laboratory conditions.The egg hatch rates were assessed 48 h post treatment.Results:The LC50 values of benzene,hexane,ethyl acetate,methanol and chloroform extract of E.alba against early third instar larvae of Ae.aegypti were 151.38,165.10, 154.88,127.64 and 146.28 ppm,respectively.Maximum larvicidal activity was observed in the methanol extract followed by chloroform,benzene,ethyl acetate and hexane extract.No mortality was observed in control.Among five solvent tested the methanol extract was found to be most effective for ovicidal activity against Ae.aegypti.The methanol extracts exerted 100%mortality （zero hatchability） at 300 ppm.Conclusions:From the results it can be concluded the crude extract of E.alba was an excellent potential for controlling Ae.aegypti mosquito.

Identification of metabolites of Radix Paeoniae Alba extract in rat bile, plasma and urine by ultra-performance liquid chromatography–quadrupole time-of-flight mass spectrometry

Ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry （UPLC-Q-TOF/MS） was developed to identify the absorbed parent components and metabolites in rat bile, plasma and urine after oral administration of Radix Paeoniae Alba extract （RPAE）. A total of 65 compounds were detected in rat bile, plasma and urine samples, including 11 parent compounds and 54 metabolites. The results indicated that glucuronidation, hydroxylation and methylation were the major metabolic pathways of the components of RPAE. Furthermore, the results of this work demonstrated that UPLC-Q-TOF/MS combined with MetaboLynx？ software and mass defect filtering （MDF） could provide unique high throughput capabilities for drug metabolism study, with excellent MS mass accuracy and enhanced MSE data acquisition. With the MSE technique, both precursor and fragment mass spectra can be simultaneously acquired by alternating between high and low collision energy during a single chromatographic run.

Radial profile of sap flow velocity in mature Xinjiang poplar(Populus alba L. var. pyramidalis) in Northwest China

Estimation of the transpiration rate for a tree is generally based on sap flow measurements within the hydro-active stem xylem. In this study, radial variation of sap flow velocity（Js） was investigated at five depths of the xylem（1, 2, 3, 5 and 8 cm under the cambium） in three mature Xinjiang poplar（Populus alba L. var. pyramidalis） trees grown at the Gansu Minqin National Studies Station for Desert Steppe Ecosystem from May to October 2011. Thermal dissipation probes of various lengths manufactured according to the Granier＇s design were installed into each tree for simultaneous observation of the radial patterns of Js through the xylem. The radial patterns were found to fit the four-parameter GaussAmp equation. The peak Js was about 27.02±0.95 kg/（dm2？d） at approximately 3 to 5 cm deep from the cambium of the three trees,and the lowest Js appeared at 1 cm deep in most of the time. Approximately 50% of the total sap flow in Xinjiang poplar occurred within one-third of the xylem from its outer radius, whereas 90% of the total sap flow occurred within two-fifth of the xylem. In addition, the innermost point of the xylem（at 8-cm depth）, which appeared as the penultimate sap flow in most cases during the study period, was hydro-active with Js,8 of 7.55±3.83 kg/（dm2？d）. The radial pattern of Js was found to be steeper in midday than in other time of the day, and steeper diurnal fluctuations were recorded in June, July and August（the mid-growing season）. Maximum differences between the lowest Js（Js,1 or Js,8） and the highest Js（Js,3 or Js,5） from May through October were 12.41, 17.35, 16.30, 18.52, 12.60 and 16.04 g/（cm2？h）, respectively. The time-dependent changes of Js along the radial profile（except at 1-cm depth） were strongly related to the reference evapotranspiration（ET0）. Due to significant radial variability of Js, the mean daily sap flow at the whole-tree level could be over-estimated by up to 29.69% when only a single probe at depth of 2 cm was used. However, the accuracy of the estimation of sap flow in Xinjiang poplar could be significantly improved using a correction coefficient of 0.885.

白环柄菇[Lepiota alba(Bresad.)Sacc]的生理学特性和栽培研究

白环柄菇是四川省菌物的一个新记录物种分类学地位为：担子菌亚门，层菌纲，伞菌目，伞菌科，环柄菇属。此菌无毒，可食。菌丝光锁状联合，形成大量菌核（Sclerotium），在伞菌目中是罕见的。详细报道了该菌的子实体、菌丝体的形态特征、生理特点和栽培技术。这是一种典型的高温性草菌。栽培出菇的培养料为草粉、牛粪、皮及食用菌栽培废料，瓶栽、袋栽、床栽均可，干菇生物学效率为4％-8％。

酸雨胁迫对白兰花(Michelia alba DC．)抗氧化酶活性及丙二醛(MDA)含量的影响

本文以长势良好且树龄相同的木兰科植物白兰花(Michelia alba.DC)为实验材料,采用盆栽方式,用模拟酸雨的处理方法,研究了其在模拟酸雨胁迫下的抗氧化酶系统和丙二醛(MDA)的变化规律。结果表明,pH2.0酸雨比pH3.0和pH4.0对白兰花的影响更大,到第六次喷施酸雨时,SOD、POD、CAT酶活性均低于对照;各处理MDA含量在第六次处理时达到最高值,且均与对照呈显著性差异(P<0.05)。说明酸雨胁迫对白兰花的影响较大,白兰花在酸雨胁迫下抗逆性和耐受性较弱,其结果为酸雨控制区选择城市绿化植物提供重要的现实意义。

不同光质对白芥(Sinapis alba)下胚轴愈伤组织形成和器官再生的影响(简报)(英文)

以连续白光下或黑暗中生长5天的白芥幼苗下胚轴切段为外植体(分别叫作L-外植体和D-外植体),在1mg/L NAA+2mg/L BA的MS培养基上诱导脱分化和芽的再分化。在白光、红光、蓝光和黑暗中培养,L-外植体都比D-外植体愈伤组织形成早,生长快;对于L-外植体形成愈伤组织的促进作用依次是:白光>红光>蓝光》黑暗;而对D-外植体蓝光比红光更有效。L-外植体比D-外植体芽再生的能力强得多,不同光质的作用为:白光>红光>蓝光>黑暗。

Monepaloside K, a New Triterpenoid Saponin from Morina nepalensis var. alba Hand. - Mazz.

One new triterpenoid saponin, monepaloside K (1) was isolated from the water-soluble part of the whole plant of a famous Tibetan medicinal herb, morina nepalensis var. alba Hand.-Mazz.. Its structure was determined to be 3-O-a-L-arabinopyranosyl-(13)-b-D- xylopyranosyl siaresinolic acid on the basis of spectroscopic evidences, especially 2D NMR techniques.

Effects of Salvia miltiorrhiza Bge.f.alba on neuronal regeneration following cerebral ischemia/reperfusion

BACKGROUND： Subsequent to cerebral ischemic injury, endogenous neural stem cells are activated, but ischemia-induced neuronal loss is not compensated by ischemic injury-induced neural regeneration. Salvia （S.） miltiorrhiza Bge.f.alba （Baihua Danshen, a Chinese herbal medicine） could enhance learning and memory functions, as well as promote neural regeneration. OBJECTIVE： To observe the effects of S. miltiorrhiza Bge.f.alba on recovery from cerebral ischemia-reperfusion injury, and the influence on neuronal regeneration and differentiation. DESIGN, TIME AND SETTING： Randomized, controlled, animal experiments were performed at the Experimental Animal Center and Neurobiology Laboratory of Taishan Medical College in September of 2006. MATERIALS： S. miltiorrhiza Bge.f.alba was provided by Taishan Medical College Botanic Garden, Taian, China; dl-3n-butylphthalide （NBP） soft capsule was purchased from NBP Pharmaceutical, Shijiazhuang, China; mouse anti-bromodeoxyuridine antibody, rabbit anti-NF200 antibody, and bromodeoxyuridine were purchased from Sigma, Louis, MO, USA; Annexin V-fluorescein isothiocyanate/PI apoptosis kit was purchased from Nanjing Comissariado Biological Technology Development, Nanjing, China. METHODS： Adult Sprague Dawley rats were randomly assigned to sham surgery, model （cerebral ischemia and reperfusion, without administration）, S. miltiorrhiza Bge.f.alba, and NBP groups. Following establishment of the cerebral ischemia/reperfusion model, S. miltiorrhiza Bge.f.alba or NBP （1 mL/100 g） was respectively perfused at 30 minutes following cerebral ischemia/reperfusion. MAIN OUTCOME MEASURES： Alterations in cerebral blood flow before and after ischemia/reperfusion, NF200- and bromodeoxyuridine-double positive cells in striatum of affected tissues, as well as neuronal apoptosis rate at days 5 and 7 following cerebral ischemia/reperfusion. RESULTS： Subsequent to cerebral ischemia reperfusion, cerebral blood flow was reduced. Following treatment with S. miltiorrhiza Bge.f.alba, cerebral blood flow significantly increased （P 〈 0.05）. NBP treatment was inferior to S. miltiorrhiza Bge.f.alba with regard to stabilization of cerebral blood flow （P 〈 0.05）. S. miltiorrhiza Bge.f.alba significantly increased the number of newly formed neurons in rats following cerebral ischemia （P 〈 0.05） and significantly reduced neuronal apoptosis （P 〈 0.05）, with no significant difference compared with NBP treatment （P 〉 0.05）. CONCLUSION： S. miltiorrhiza Bge.f.alba significantly increased cerebral blood flow, reduced neuronal apoptosis, and promoted neuronal regeneration in rats with cerebral ischemia/reperfusion impairment.

Effects of extracts from Hibiscus macranthus and Basella alba mixture on testosterone production in vitro in adult rat testes slices

Aim： To find an in vitro system for the measurement of the androgenic effects of different extracts of Hibiscus macranthus （Malvaceae） and Basella alba （Basellaceae）. Methods： The production of testosterone from testes slices incubated in two media, either Krebs-Henseleit buffer containing 0.5% Bovine serum albumin （BSA） or Dubecco＇s Modified Eagle＇s medium-F12 Ham nutrient mixture （DME/Ham F12）, under a mixture of 5% CO2 in 95% air was determined either in the presence or absence of cofactors and Hibiscus macranthus plus Basella alba （HMBA） extracts. Results： The testosterone production was increased in testes slices incubated in DME/Ham F12 medium in response to the cofactors （49%） and aqueous extracts （34%-60% according to dilutions）. Under the same atmospheric conditions, there was no positive response of the testes slices to either cofactor or HMBA extract stimulation in Krebs-Henseleit buffer containing 0.5% BSA, In further investigations related to the effect of HMBA, the DME/ Ham F12 medium was used. The results obtained from the in vitro test showed that the activity was present mainly in methylene chloride and methanol, since these extracts induced an increase in testosterone production by testes slices. Conclusion： The testes slice system is suitable to be used for further in vitro investigations of the isolation of androgenic bioactive components of plants. （Asian J Androl 2006 Jan; 8： 111-114）

A new alkylene dihydrofuran glycoside with antioxidation activity from the root bark of Morus alba L.

A new alkylene dihydrofuran glycoside（1） was isolated from the root bark ofMorus alba L.,along with moracin M-3′-O-β-D-glucopyranoside （2）,and moracin M-6,3′-di-O-β-D-glucopyranoside（3）.Compound 1 was identified as 2-methylene-3-methoxy-2, 5-dihydrofuran-4-O-β-D-glucopyranoside on the basis of chemical and spectroscopic data including 1D and 2D NMR spectral analysis.In addition,the antioxidant activity of 1 was evaluated using the 1,1-diphenyl-2-picrylhydrazyl（DPPH） and 2,2′-azinobis- 3-ethylbenzothiazoline-6-sulphonic acid（ABTS） assay.The IC_（50） values were 2.49 and 0.45 mg/mL,respectively.

Effects of pruning intensity on nonstructural carbohydrates of Populus alba×P.talassica in the arid desert region of Northwest China

Although pruning is important to obtain highquality,large-diameter timber,the effects of pruning on nonstructural carbohydrates(NSC)in aboveground organs of many timber species are not well understood.Three intensities of pruning(none,moderate and severe)were tested on poplars(Populus alba×P.talassica)in the arid desert region of northwest China to compare the concentrations of soluble sugar(SS),starch(ST)and total nonstructural carbohydrate(TNC)in leaves,branches and trunks during the growing season.The concentration of NSC components after different pruning intensities varied similarly in seasonal patterns,increasing slowly at the beginning of the growing season,continuously declining in the middle,then gradually recovering by the end of the growing season.The monthly mean NSC concentration in poplar differed significantly among the three pruning intensities(p<0.05).The SS concentration in pruned trees was higher than in unpruned trees(p<0.05).For moderately pruned trees,the concentrations of ST and TNC in trunks and branches were higher than in unpruned and in severely pruned trees(p<0.05).Compared with no pruning,pruning changed the seasonal variation in NSC concentration.The orders of SS and TNC concentrations in aboveground organs were leaf>branch>trunk,while the order of ST concentration was trunk>leaf>branch,which was related to functional differences of plant organs.The annual average growth in height of unpruned,moderately pruned,and severely pruned poplars was 0.21±0.06,0.45±0.09 and 0.24±0.05 m,respectively,and the annual average growth in DBH were 0.92±0.04,1.27±0.06 and 1.02±0.05 cm,respectively.Our results demonstrate that moderate pruning may effectively increase the annual growth in tree height and DBH while avoiding damage caused by excessive pruning to the tree body.Therefore,moderate pruning may increase the NSC storage and improve the growth of timber species.

当年生桑树(Morus alba L.)果枝叶片特征与果实性状的相关性

为揭示叶果间的内在作用规律,以桑树(Morus alba L.)当年生果枝为研究对象,分析不同大小果枝上叶片特征和果实性状的差异及其二者间的相互关系。结果表明:随着果枝由小到大,果枝上的叶片数与总叶干重显著增加,而叶形指数却无显著变化;桑葚数与总桑葚干重也有显著增加,桑葚含水量无显著变化。桑葚数与叶片数、总叶干重、总叶面积及其平均桑葚干重显著正相关。研究结果证实了果实的性状与叶片特征之间存在极密切关系。当年生果枝上的桑葚数量多少取决于果枝上的叶片总数,平均桑葚的干重则与果枝上的平均叶面积显著相关。

Research and Prospect of Mulberry (Morus alba L.)

Mulberry(Morus alba L.)is not only rich in nutrients,but also has medicinal value.As a drug and food,it is widely planted all over the country,with considerable yield and economic value.In recent years,it has attracted many scholars and been widely reported,and in-depth research has already been conducted.Through reviewing the literatures on mulberry,this paper summarized the chemical composition,pharmacological effects and quality evaluation of mulberry,providing some reference for the further development and utilization of mulberry.

In vitro antimicrobial activity of mangrove plant Sonneratia alba

Objective:To investigate the antimicrobial property of mangrove plantSonneratia alba(S. alba).Methods:The antimicrobial activity was evaluated using disc diffusion and microdilution methods against six microorganisms. Soxhlet apparatus was used for extraction with a series of solvents,n-hexane, ethyl acetate and methanol in sequence of increasing polarity.Results:Methanol extract appeared to be the most effective extract whilen-hexane extract showed no activity. The antimicrobial activities were observed against the gram positive bacteria Staphylococcus aureus(S. aureus) and Bacillus cereus(B. cereus), the gram negative Escherichia coli(E. coli) and the yeast Cryptococcus neoformans. Pseudomonas aeruginosa and Candida albicans appeared to be not sensitive to the concentrations tested since no inhibition zone was observed.E. coli(17.5 mm) appeared to be the most sensitive strain followed by S. aureus(12.5 mm)and B. cereus(12.5 mm).Conclusions:From this study, it can be concluded that S. alba exhibit santim icrobial activities against certain microorganisms.

Gene cloning and expression analysis of limonene synthase in Syringa oblata and S.oblata var.alba

Syringa species are important ornamentals with strong floral scent,of which monoterpenes are the main component.In this study,a new monoterpene synthase gene,named SoLIM,was collected from the flowers of Syringa oblata and S.oblata var.alba using a homologous cloning method.The full-length cDNA of SoLIM was1746 bp and encoded 581 amino acids.Sequence analysis showed that SoLIM contained the DDxxD and RRx8 W motifs,which are two typical conserved monoterpene synthase motifs,and was thus classified as belonging to the Tpsb subfamily.Using quantitative reverse-transcription PCR,SoLIM was significantly expressed in the petals and pistils of S.oblata and S.oblata var.alba,respectively.SoLIM expression peaked earlier than the D-limonene emissions in the diurnal experiments,but occurred later when D-limonene had peaked during the flowering phase,indicating that differences in SoLIM gene expression and D-limonene emissions existed.The synthesis of floral scent is thus associated with diverse regulatory mechanisms that require further investigation.