Research progress on chemical constituents in Sophora alopecuroides L.and their pharmacological activities

Sophora alopecuroides L.,a perennial plant belonging to Leguminosae family,is used as medicinal materials to treat a variety of skin diseases.Its chemical components are complex and diverse,mainly including alkaloids and flavonoids.Modern pharmacological studies have found that the isolated compounds from S.alopecuroides have a variety of pharmacological activities.In particular,alkaloids represented by matrine,aloperine and oxymatrine have significant anti-inflammatory,anti-viral,anti-tumor,anti-bacterial and immunomodulatory activities.This paper summarized the chemical constituents and pharmacological activities of S.alopecuroides,aiming to provide the basis for further research.

Sophaloseedlines A-G: Diverse Matrine-Based Alkaloids from Sophora alopecuroides with Potential Anti-Hepatitis B Virus Activities

Sophaloseedlines A-G(1-7),seven new matrine-based alkaloids along with two known analogues,were isolated from the seeds of Sophora alopecuroides.The new structures were determined based on extensive spectroscopic data,electronic circular dichroism calculations,and X-ray crystallography.Notably,sophaloseedline A(1)represents a novel rearranged 6(5→17)-abeo-matrine alkaloid featuring unprecedented highly constructed 7/6/5/6 tetracyclic fused ring skeleton.The hypothetical biosynthetic pathways for sophaloseedli nes A-F were proposed based on co-existing precursors.Additionally,all the isolated alkaloids were screened for their antiviral activities against hepatitis B virus,and new alkaloids 1 and 2 displayed more potent activities than those of matrine(a parent alkaloid of title plant)and positive control(lamivudine).

Structurally Diverse Matrine-Based Alkaloids with Anti-inflam-matory Effects from Sophora alopecuroides

A phytochemical investigation on the seeds of Sophora alopecuroides led to obtaining fourteen structurally diverse matrine-based alkaloids(1-14),including eight new ones(1,6,8-12,14).Notably,alopecuroide F(1)represents the first dimeric matrine-type skeleton assembled via unprecedent C-13-C-12'connection.The new structures were determined through extensive spectroscopic data(UV,OR,HRESIMS,1D,and 2D NMR),ECD calculations,and three instances,verified by X-ray crystallography.Biologically,all alkaloids were evaluated for cytotoxicity against four human cancer cell lines(HepG2,A549,THP-1,and MCF-7)and anti-inflammatory activities for two pro-inflammatory cytokines(TNF-αand IL-6).Alopecuroide F(1)can inhibit TNF-αand IL-6 productions in a dose-dependent manner with IC50 values of 35.6±0.5 and 41.7±0.8μmol/L,respectively.

Anti-helicobacter pylori effect of total alkaloids of sophora alopecuroides in vivo

Background Helicobacterpylori （H.pylori） infection could lead to most gastroduodenal diseases and is even identified as a carcinogen of gastric cancer.Total alkaloids of sophora alopecuroides （TASA） is widely used in herbal remedies to treat various infectious diseases,including stomach-associated diseases.This study is aimed at evaluating the antimicrobial activity of TASA on H.pylori-infected BALB/c mice mouse gastritis.Methods Totally 120 BALB/c mice were orally inoculated with H.pylori Bacterial liquid to construct BALB/c mice H.pylori infection gastritis animal model,after the model was successfully created.We randomly assigned 100 infected mice into 10 treatment groups,the first group （normal saline）; the second group （bismuth pectin）; the third group （omeprazole）; the fourth group （TASA 2 mg/d）; the fifth group （TASA 4 mg/d）; the sixth group （TASA 5 mg/d）; the seventh group （TASA ＋ bismuth pectin）; the eighth group （TASA ＋ omeprazole）; the ninth group （bismuth pectin ＋ clarithromycin ＋ metronidazole）;the tenth group （omeprazole ＋ clarithromycin ＋ metronidazole）,5 other non-infected mice as negative control.Mice were orally inoculated twice a day and 7 days continuously.Then the mice were killed 4 weeks after treatment,we used realtime PCR to detect 16sDNA of H.pylori to test both the colonization and the clearance mice of bacteria of each treatment.We applied hematoxylin and eosin （HE） staining and immunostaining of mice gastric mucosa to observe the general inflammation and related factors interleukin 8 （IL-8）,cyclooxygenase 2 （COX-2）,and nuclear factor-kappa B （NF-KB） expression change after treatments.Results Firstly,we ensured that after 6-week intragastric administration,the bacteria colonization reached an exceed peak which is far higher than positive threshold （P ＜0.001）; secondly,after treatments,it is revealed that TASA combined with omeprazole or bismuth pectin showed promising antimicrobial activity against H.pylori as well as conventional triple therapy （P ＜0.001）; thirdly,HE staining showed that the inflammation on mice gastric mucosal membrane were also relieved obviously in TASA combined treatments and conventional triple therapy compared with normal saline treated mice,moreover,from immunohistochemistry results,H.pylori-induced IL-8,COX-2,and NF-KB were consistently suppressed in seventh,eighth,ninth,and tenth group to a certain extent.Conclusion These results open the possibility of taking TASA as an anti-inflammatory agent for H.pylori gastritis.

Protective Effect of Total Alkaloids of Sophora Alopecuroides on Dextran Sulfate Sodium-Induced Chronic Colitis

Objective：To investigate the effect of total alkaloids of Sophora alopecuroides（TASA） on dextran sulfate sodium（DSS）-induced colitis in mice.Methods：Chronic experimental colitis was induced by administration of 4 cycles of 4%DSS.Fifty mice were randomly distributed into 4 groups（normal,DSS,DSS/high-dose TASA, and DSS/low-dose TASA groups） by a random number table with body weight stratification.Mice in the normal group（n=11） and DSS-induced colitis control group（n=15） received control treatment of 20 mL/kg distilled water; DSS plus TASA high- and low-dose groups（n=12 each） were treated with TASA solution（20 mL/kg） at the doses of 60 mg/kg and 30 mg/kg,respectively.The severity of colitis was assessed on the basis of clinical signs, colon length,and histology scores.Moreover,secretory immunoglobulin A（slgA） and haptoglobin（HP） were analyzed by enzyme linked immunosorbent assay;intercellular adhesion molecule 1（ICAM-1） and macrophage-migration inhibitory factor（MIF） gene expressions were analyzed by quantitative reverse transcriptase realtime polymerase chain reaction（qRT-PCR） using SYBA greenⅠ;and nuclear factorκB（NF-κB） expression and activation and p65 interaction with the promoter of ICAM-1 gene were assessed by Western blotting and chromatin immunoprecipitation assay.Results：TASA administration significantly attenuated the damage and substantially reduced HP elevation and maintained the level of cecum slgA.TASA inhibited the ICAM-1 gene expression and had no effect on MIF gene expression.Also,TASA was able to reduce phospho-lκBα（p-lκBα） protein expression;however,it had no effect on the activation of IκB kinaseα（IKKα） and inhibitor of NF-κBα（IκBα）.Moreover,TASA inhibited the p65 recruitment to the ICAM-1 gene promoter.Conclusions：TASA had a protective effect on DSS-induced colitis.Such effect may be associated with its inhibition of NF-κB activation and blockade of NF-κB-regulated transcription activation of proinflammatory mediator gene.

Preparation of Colon‐specific and Synchronous Release Pellet Containing Total Alkaloids of Sophora alopecuroides

Objective To deliver multiple component drugs to colon site and sustain a synchronous release for better therapeutic effect. For achieving this purpose, colon specific pellet containing total alkaloids of Sophora alopecuroides （TASA） was prepared. Methods The pellet was prepared by extrasion-spheronizing and subsequently coated with three layers of two polymers. Results The pellet core consisted of 40% TASA, 1：2 in ratio of Bletilla striata polysaccharide （BSP）, an enzyme-degradable material, to microcrystalline cellu{ose （MCC）, filler, and 1% CMC-Na solution as binder by optimization. Concerning of the three coated layers, the outer layer was coated with Eudragit RS30D for controlling drug release in colon, the intermediate layer and the inner layer were coated with same polymer, Eudragit $100, for preventing drug release in upper gastrointestinal tract, which required 23.2%, 21.7%, and 9.3% weigh gain, respectively. The coated pellets released 1.20% of sophoridine and 1.98% of matrine in media mimicking the stomach condition for 2 h, and 23.88% of sophoridine and 22.91% of matrine in media mimicking the intestine for 3 h and finally 90.25% of sophoridine and 89.94% of matrine in colonic conditions within 24 h, And the similarity factor f2 of sophoridine and matrine of release curve for investigated formulation was internal in （50-100） and 〉 80, demonstrating that sophoridine and matrine in formulation achieved a synchronous release. Conclusion The coated pellets achieve a certain colon-specific release and synchronous release.

Simultaneous Determination of Multiple Bioactive Constituents in Total Alkaloid of Sophora alopecuroides by HPLC-DAD

Objective To develop a qualitative and quantitative simultaneous determination of multiple bioactive constituents in total alkaloid in Sophora alopecuroides(TASA).Methods In the experiment,a new and simple HPLC-DAD method for the simultaneous determination of multiple constituents in TASA was developed.The separation was performed on a Kromasil C18 column(250 mm×4.6 mm,5.0μm)eluted with 0.02 mol/L potassium dihydrogen phosphate(adjusted pH 4.3 using 1%glacial acetic acid)and acetonitrile(75:25)at a flow-rate of 0.7 mL/min.The detection wavelength was set at 210 nm.Results Five constituents(sophoridine,matrine,oxymatrine,aloperine,and lehmannine)were simultaneously analyzed in this study.Four of them were identified and determinated by the developed method.The calibration curves exhibited linear regressions(r 2 >0.9995).The injection precision,the intra-day precision,and the analysis repeatability were validated with the RSD values less than 5.0%.The mean recoveries of the four constituents were ranged from 98.62%to 100.20%,and the RSD values were all less than 3.37%.Conclusion This method is convenient,fast,accurate,and is applicable to analyze the multi-constituents in TASA.

Total alkaloids of Sophora alopecuroides-and matrine-induced reactive oxygen species impair biofilm formation of Staphylococcus epidermidis and increase bacterial susceptibility to ciprofloxacin

Objective:To investigate the mechanism by which total alkaloids of Sophora alopecuroides(TASA)and matrine(MT)impair biofilm to increase the susceptibility of Staphylococcus epidermidis(S.epidermidis)to ciprofloxacin.Methods:The minimum biofilm inhibitory concentration(mBIC)was determined using a 2-fold dilution method.Structure of biofilm of S.epidermidis was examined by Confocal Laser Scanning Microscope(CLSM).The cellular reactive oxygen species(ROS)was determined using a DCFH-DA assay.The key factors related to the regulation of ROS were accessed using respective kits.Results:TASA and MT were more beneficial to impair biofilm of S.epidermidis than ciprofloxacin(CIP)(P<0.05).TASA and MT were not easily developed resistance to biofilm-producing S.epidermidis.The mBIC of CIP decreased by 2-6-fold following the treatment of sub-biofilm inhibitory concentration(sub-BIC)TASA and MT,whereas the mBIC of CIP increased by 2-fold following a treatment of sub-BIC CIP from the first to sixth generations.TASA and MT can improve the production of ROS in biofilmproducing S.epidermidis.The ROS content was decreased 23%-33%following the treatment of submBIC CIP,whereas ROS content increased 7%-24%following treatment with TASA+CIP and MT+CIP combination from the first to sixth generations.Nitric oxide(NO)as a ROS,which was consistent with the previously confirmed relationship between ROS and drug resistance.Related regulatory factorssuperoxide dismutase(SOD)and glutathione peroxidase(GSH)could synergistically maintain the redox balance in vivo.Conclusion:TASA and MT enhanced reactive oxygen species to restore the susceptibility of S.epidermidis to ciprofloxacin.

Alopecuroidines A-C, three matrine-derived alkaloids from the seeds of Sophora alopecuroides

Three matrine-derived alkaloids, alopecuroidine A(1), alopecuroidine B(2 a) and alopecuroidine C(2 b)were isolated from the seeds of Sophora alopecuroides. Their structures were elucidated by extensive spectroscopic analyses and X-ray diffraction. Three compounds possess an unprecedented rearranged fused7/6/5/6 tetracyclic skeleton with a diazacycloheptane structure. Their plausible biosynthetic pathway was also proposed. The anti-proliferative activities of compounds 1 and 2 a were examined by the MTT assay.Compound 1 inhibited the viability of human lung cancer A549 cells, having a half maximal inhibitory concentration(IC50) of 7.58 ± 2.47 μmol/L at 72 h. The flow cytometric analysis suggested that 1 inhibited A549 cell growth by inducing apoptosis and cell-cycle arrest. Additionally, 1 induced the loss of mitochondrial membrane potential, elevated intracellular reactive oxygen species, increased the Bax/Bcl-2 ratio, stimulated cleaved-caspase-3 and P53 protein levels, and suppressed the pro-caspase-3 level. Thus,1 appeared to induce A549 cells apoptosis through a mitochondria-mediated apoptotic pathway.

Three quinolizidine dimers from the seeds of Sophora alopecuroides and their hepatoprotective activities

Sophoralines A–C,three novel[2+2]cycloaddition dimers of matrine-based alkaloids with an unprecedented 6/6/6/6/4/6/6/6/6 nonacyclic skeleton containing 11 stereogenic centers,were isolated from Sophora alopecuroides.Their structures were determined by spectroscopic methods,and the absolute configurations were further determined by single-crystal X-ray diffraction analysis for 1 and quantum chemical calculations of electronic circular dichroism(ECD)spectra for 2 and 3.Moreover,1 exhibited excellent hepatoprotective activities in acetaminophen-induced liver injury in vitro and in vivo.

Genetic Diversity of Antagonistic Endophytic Bacteria Isolated from Sophora alopecuroide

[ Objective ] The paper was to study and analyze the genetic diversity of 48 endophytic bacteria isolated from Sophora alopecuroide with strong antagonis- tic effect against Fusarium oxysporum and Verticillium dahliae, and carry out sequence measurement and phylogenetic analysis on 10 representative strains. [ Meth- od] Using total DNA ERIC-PCR and 16S rRNA sequencing analysis method, a total of 48 endophytic bacteria were carried out genetic diversity and phylogenetic analysis. [Result] ERIC-PCR fingerprint map showed that 48 endophytic bacteria were divided into 6 ERIC groups and 2 strains with independent groups at Wat- son distance of 0.31. Representative strains were selected from each group to determine 16S rRNA gene for phylogenetic analysis, and the results showed that these strains belonged to Bacillus atrophaeus, Bacillus subtilis, Bacillus cereus, Bacillus megaterium, Bacillus pumilus and Serratia marcescens, respectively. [ Conclu- sion ] The genetic diversity of 48 strains of bacteria was obvious. Determination of 48 strains of bacteria on diversity and phylogenetic status could lay foundation for study on mechanism of F. oxysporum and V. dahliae, which could also provide new strain sources for biological control of diseases in cotton production.

苦豆子绿肥的腐解和养分释放研究

以营养期至初花期苦豆子(Sophora alopecuroides)为试验材料,用尼龙袋法研究苦豆子绿肥在土壤中的腐解规律和C、N、P、K养分的释放规律。结果表明,经过118 d的腐解,绿肥中N、P矿化率逐步增加,C矿化率为66.59%;K在0~7 d矿化迅速,矿化率为55.46%,至118 d共矿化79.06%。C/N在0~14 d呈下降趋势,14~118 d逐渐平稳,养分矿化率表现为K>C>P>N。在118 d苦豆子绿肥干物质(DM)腐解量达65.2%,C、N、P、K矿化率分别达62.75%、44.93%、52.54%和79.06%,其中,0~14 d为快速分解阶段、14~28 d为减慢分解阶段、28~118 d为缓慢分解阶段。苦豆子绿肥的干物质降解与养分释放基本同步,且呈正相关。

Five quinolizidine alkaloids with anti-tobacco mosaic virus activities from two species of Sophora

Three novel matrine-type alkaloids(1-3)and two unprecedented aloperine-type alkaloids(4 and 5)were isolated from the root of Sophora tonkinensis and the seeds of Sophora alopecuroides respectively.Notably,compound 1 possessed an unprecedented 6/5/6 tricyclic skeleton,while compounds 2 and 3 characterized by rare 6/6/5/6 tetracyclic system and 6/6/6/6/6 pentacyclic system respectively.Moreover,compound 4 possessed an unprecedented 6/7/6/6 tetracyclic core,and compound 5 characterized by rare 6/6/6/6tetracyclic skeleton.Their structures were elucidated by comprehensive spectroscopic data analysis and electronic circular dichroism(ECD)calculations.Biological tests indicated that compound 5 displayed significant anti-tobacco mosaic virus(TMV)activity compared with the positive control ningnanmycin.

information

The 3rd International Conference and Exposition on the Modernization of Traditional Chinese Medicine was held during Nov.25-26,2010 in Chengdu(Sichuan,China),editorial director CHEN Chang-qing associated with other members of Chinese Herbal Medicines(CHM)editorial office attended this meeting.