Synergistic effect of pinellia total alkaloids and uncaria total alkaloids on anticonvulsant action in mice and rats

Aim To investigate the synergistic effect of the combination of pinellia total alkaloid （PTA） and uncaria total alkaloid （UTA）, and explore the mechanism of anticonvulsant action. Methods Anticonvulsant and toxic effect profiles of combinations of PTA with UTA, alone and at three fixed ratios of 1：4, 1 ：1, 4：1, were evaluated in maximal electroshock （MES）-induced seizures and acute toxicity test in mice. Respective ED50 and LD50 were calculated with Bliss＇s method. Their synergistic effect were evaluated by isobolographic analysis and allowed the determination of benefit indices （BI） for respective combinations. The model of convulsive rats kindled by penicillin topically injected into cortex was used to investigated the content of Glu, Asp, Gly and GABA in hippocampus using high performance liquid chromatography （HPLC）. Results Combinations of PTA and UTA at the ratio of 4：1 were synergistic in MES test and antagonistic in acute toxicity test, showing the best profile for combinations of PTA with UTA. In contrast, the ratios of 1 ：4 and 1 ： 1, despite synergistic in MES test, were additive in acute toxicity test. The 4：1 combination and two drugs alone significantly decreased Glu level and increased GABA level in the hippocampus, but the GABA level in the 4：1 combination group was higher than that in the two drugs alone groups. They did not have significant influence on the levels of ASp and Gly. Conclusion Combinations of PTA and UTA at 4：1 ratio demonstrated synergistic effect in anticonvulsant action and antagonistic effect in toxicity. The anticonvulsant mechanism might be related to decreasing the excitability of Glutamatergic neurons and increasing the inhibition of GABAergic neurons.

Determination of the Content of Total Alkaloids in Different Solvent Extracts from Climbing Groundsel Herb Produced in Guizhou Province and Their Antibacterial Activity

[Objective] This study aimed to compare the contents of total alkaloids in different solvent extracts from climbing groundsel herb produced in Guizhou Province and their antibacterial activity. [Method] The content of total alkaloids was measured by UV spectrophotometry, and Oxford cups were used to investigate the antibacterial activity of each solvent extract. [Result] There was a good linear correlation between the absorbance measured by the UV spectrophotometer and the content of total al- kaloids within the concentration of 0.011 0-0.054 8 mg/ml at 207 nm, and the re- gression equation was Y=23.654X＋0.021, R=0. 999 7 and the average recovery rate was 99.2%. The contents of total alkaloids in 60% ethanol extract, 95% ethanol and water ethanol were 38.71, 52.25 and 60.50 mg/g, respectively. The 60% ethanol ex- tract had strong antibacterial activity against Streptococcus pneumoniae, Staphylo- coccus aureus and Escherichia coil; the water extractive had stronger antibacterial activity against Escherichia colr, 95% ethanol extract had weak antibacterial activity, with no inhibition of Pseudomonas aeruginosa. [Conclusion] This method is accurate, simple, with good repeatability, which can be used for the determination of alkaloids content of the climbing groundsel herb; there is no positive correlation between the content of total alkaloids in different solvent extracts of climbing groundsel herb and their antibacterial activity.

Study of Total Alkaloids from Rhizoma Coptis Chinensis on Experimental Gastric Ulcers

Objective： To study the effects of total alkaloids （TA） extracted from Rhizoma Coptis Chinensis on experimental gastric ulcer models. Methods： Four kinds of experimental ulcer models were established respectively by water-immersion stress, intragastric ethanol, acetic acid erosion, and pylorus ligation. The anti-ulcer effects of TA were evaluated, and compared with that of berberine （Bet） and cimetidine （Oim）. Results： TA showed significant inhibitory effects on ulcerative formation induced by water-immersion stress, intragastric ethanol, and pylorus l igation in dose-dependent manner, and showed therapeutic effect on acetic acid erosion-inducing ulcer, in comparison with the control group. The anti-ulcer activity of Bet was less than TA containing equal content of Bet. TA significantly reduced the free acidity, total acidity and total acid output, but didn＇t affect the gastric juice volume, gastric pepsin activity, adherent mucus quantity of stomach wall and free mucus dissolving in gastric juice. The suppressive activities of TA on gastric acid secretion didn＇t occur when it was administered into dodecadactylon at a dose of 360 mg/kg wt. Moreover, when compared with Oim, the inhibitory effect of TA on gastric acid secretion isn＇t proportional to the inhibitory effects on the formation of the 4 kinds of experimental ulcers. Conclusion： TA is a potent candidate in therapeutic drugs for treating gastric ulcer. Its anti-ulcer effective components and mechanism is not only related to Bet and inhibition of gastric acid, but also to other ingredients of TA and mechanism so far unknown.

Analysis of Hupehenine in the Total Alkaloids from Fritillaria hupehensis by HPLC-ELSD

A reverse-phase high pressure liquid chromatography （HPLC） method with evaporative light scattering detection （ELSD） has been developed for the quantitative analysis of hupehenine in the total alkaloids from Fritillaria hupehensis. Samples were analyzed on a reverse-phase column （Hypersil C-18） with a mobile phase of methanol：water：chloroform： triethylamine （85：15：1：0.6）. The ELSD was set at the drift tube temperature of 68.3℃ and gas flow rate of 1.8 L/min. Hupehenine＇s retention time was 13.7 min with an asymmetry factor of 1.2. The validity of the method has been verified with linearity, limit of detection, accuracy and precision. The logarithmic linear curve was obtained from 8.936 to 134.04 μg/mL （r=0.9993）. The detection limit （S/N〉3） of hupehenine was 1.79 μg/mL on the column. Intra-day RSD was 1.42% and inter-day RSD was 2.26% （3 days within a week）. The average recovery of hupehenine was 101.50%, and RSD was 1.62%.

Antihyperglycemic and antioxidant activities of total alkaloids from Catharanthus roseus in streptozotocin-induced diabetic rats

We evaluated the hypoglycemic and antioxi- dant effects of the total alkaloids of leaves and twigs of Catharanthus roseus Linn. （CTA） in streptozotocin （STZ）- induced diabetic rats. The hypoglycemic effect was mea- sured by blood glucose and plasma insulin level. Oxidative stress was measured in heart, liver and kidney by levels of antioxidant markers, free radical scavengers and lipid peroxides i.e. superoxide dismutase （SOD）, catalase （CAT）, glutathione （GSH） and thiobarbituric acid reactive substances （TBARS）. Biochemical parameters, i.e. aspar- tate aminotransferase （AST）, alanine aminotransferase （ALT）, alkaline phosphate （ALP） were observed in diabetic control and treated rats. Oral administration of CTA for 30 days was followed by a significant （P 〈 0.05） decrease in fasting blood glucose and increase in insulin level as compared with untreated diabetic rats. Also it significantly （P 〈 0.05） reduced ALT, AST and ALP. The treatment also resulted in significant （P 〈 0.05） reductions in GSH, SOD, CAT, and decrease in TBARS in the heart, liver and kidney of diabetic rats. The results suggest that CTA can effectively normalize the impaired antioxidant status in STZ-induced diabetes in a dose-dependent manner.CTA exerted rapid protective effects against lipid peroxi- dation by scavenging of free radicals and reducing the risk of diabetic complications.

STUDIES ON GLYCOSIDES ⅤⅢ.TOTAL SYNTHESIS OF NEW ALKALOID GLYCOSIDES FROM SELAGINELLA DOEDERIENII

Two new alkaloids,hordenine-O- -L-rhamnopyranoside(Ⅰ)and hordenine -0-(6′-trans-cinnamoyl)-4′-O-β-D-glucopyranosyl- -L-rhamnopyranoside(Ⅲ) isolated from Selaginella doederleinii have been synthesized the first time. A new method,in which trifluoroacetoxyl group was used both as a good leaving group and a protecting group for synthesis of O-glycosides with high stereo- selectivity was reported.

Cytotoxic activity and phytochemical standardization of Lunasia amara Blanco wood extract

Objective: To evaluate the cytotoxic activity of wood extracts of Lunasia amara Blanco(L. amara) and to perform further phytochemical standardization.Methods: The wood extracts of L. amara were assessed for cytotoxic activity by in vitro tetrazolium bromide(MTT) method against two human cancer cell lines, cervical cancer cells(He La) and breast cancer cells(T47D). Thin layer chromatography, Dragendorf,acetic anhydride-sulfuric acid and ferric chloride were used to detect alkaloids, steroids and polyphenols, respectively. Furthermore, quantitative determination of total alkaloid by ultra fast liquid chromatography-photodiode array detection using lunacrine as a marker compound was performed as well.Results: The ethyl acetate extract exhibited higher cell-growth inhibition than methanol and n-hexane extracts on He La and T47 D cancer line cells with the IC50 of 71.15 and79.04 mg/m L, respectively. Total alkaloid in ethyl acetate extract was counted as(10.46 ± 0.28)%(w/w), while lunacrine determined by ultra fast liquid chromatographyphotodiode array detection method was found to be(3.55 ± 0.26)%(w/w).Conclusions: The high total alkaloid and lunacrine concentration on the extract confirm the potential cytotoxic property of ethyl acetate wood extract of L. amara.

Anti-microbial Effects In Vitro and In Vivo of Alstonia scholaris

Alstonia scholaris could be used as a traditional medicinal plant in China for the treatment of acute respiratory,which might be caused by respiratory tract infections.The investigation tested the anti-infective effects of total alkaloids extract(TA)from leaves of A.scholaris,and as a result,TA inhibited herpes simplex virus type 1(HSV-1),respiratory syncytial virus(RSV)and influenza A virus(H1N1)in vitro respectively.In addition,the survival days of mice were prolonged,and the lung weights and mortality of mice were decreased significantly,after oral administrated TA in H1N1 and beta-hemolytic streptococcus infectious models in vivo respectively.The finding supported partly the traditional usage of A.scholaris in the treatment of respiratory infections.

The effects of pinellia total alkaloids on the amino acid concentration and the GABAA receptor expression in hippocampus region of epileptic rats

The antiepileptic effect of pinellia total alkaloids（PTA） on penicillin（PNC） chronically kindled rats was investigated. We investigated the effects of PTA on Glu,Asp,Gly andγ-aminobutyric acid（GABA） concentrations and the expression level of cerebral GABA_A receptor in hippocampus.The influence of PTA on epilepsy seizure latency and degree in PNC chronically kindled rats were observed.High performance liquid chromatography（HPLC） was adopted to measure the concentrations of Glu, Asp,Gly and GABA in hippocampus. Reverse transcriptase-polymerase chain reaction （RT-PCR） was used to determine the expression of cerebral GABAA receptor mRNA. Compared with normal rats, the levels of GABA and Gly decreased obviously while the level of Glu and Asp increased significantly in model rats. The cerebral GABAA receptor mRNA level was also decreased at the same time. The difference was statistically different compared to the control group. PTA could prolong the latent period of the penicillin induced seizure and weaken the extent of seizure, compared with the model group without PTA treatment. Moreover, PTA increased the level of GABA and the expression level of GABAA receptor, while decreased the level of Glu significantly. However, it had no obvious effect on the level of Gly and Asp. Pre-treatment of PTA can also increase the GABAA receptor mRNA level. In conclusion, PTA could alleviate the PNC chronically kindled rat seizure. It increased the GABA level and the expression of GABAA receptor, and it decreased the Glu concentration.

Pinellia Total Alkaloids Modulate the GABAergic System in Hippocampal Formation on Pilocarpine-Induced Epileptic Rats

Objective:To investigate the neuromodulatory effect of pinellia total alkaloids(PTA)on the gamma-aminobutyric acidergic(GABAergic)system in epileptic rats,and preliminarily evaluate the anti-epileptic effect of PTA.Methods:Ninety-one male Sprague-Dawley rats were randomized to a control group(n=17)or an epileptic group(n=74)using computer-generated random numbers.Status epilepticus(SE)was induced with pilocarpine in the epileptic group.Epileptic rats that survived SE were randomly divided into 4 groups,namely an epilepsy group(n=13),a topiramate(TPM,60 mg/kg)group(n=12),a high-dose PTA(800 mg/kg)group(n=12),and a low-dose PTA(400 mg/kg)group(n=10).Treatments were given intragastrically once daily for 14 days.The control group and epilepsy group received normal saline.Spontaneous recurrent seizures(SRSs)were monitored 8-h daily for 7 days after treatment.Then,the hippocampal formation tissues were collected.GABA level was measured using enzyme-linked immunosorbent assay.Protein and mRNA expression levels of glutamate decarboxylase 65(GAD65),GABA transporter-1(GAT-1),GABA transaminase(GABA-T),and GABAA receptor(GABAAR)α4,α5,γ2 andδsubunits were measured using Western-blotting analysis and quantitative polymerase chain reaction.Results:PTA lowered the incidence and frequency of SRS(both doses vs.the TPM group,P>0.05).Compared with the epilepsy group,PTA increased the levels of GABA(both doses P<0.01)and GAD65(mRNA,800 mg/kg,P<0.01),and suppressed the levels of GAT-1(mRNA,800 mg/kg,P<0.01;400 mg/kg,P<0.05),GABA-T(mRNA,both doses P<0.01),and GABAARδsubunit(protein,800 mg/kg,P<0.05)andγ2 subunit(protein,both doses P<0.01).PTA upregulated the low-expressed mRNA levels of GABAARα5 subunit(400 mg/kg,P<0.01),δsubunit(800 mg/kg,P<0.05),andγ2 subunit(400 mg/kg,P<0.05).Conclusions:PTA regulated the GABAergic system through modulating GABA levels and the expression levels of GAD65,GAT-1,GABA-T,and GABAARα4,α5,γ2 andδsubunits.PTA may exert antiepileptic effects on the pilocarpine-induced epilepsy model.

Total Alkaloids from Sophora Alopecuroides L.Increase Susceptibility of Extended-Spectrum β-Lactamases Producing Escherichia coli Isolates to Cefotaxime and Ceftazidime

Objective： To evaluate the antimicrobial activity of total alkaloids extracted from Sophorea alopecuroides L. （TASA） against clinical isolated extended-spectrum beta-lactamases （ESBLs） producing Escherichia coil （E.. coil） strains. Methods： The antibacterial activity of TASA either itself or in combination with cefotaxime （CTX） or ceftazidime （CAZ） was investigated by using the microbroth dilution method and phenotypic confirmatory disk diffusion test against three clinical isolated ESBLs-producing E. coil strains; the interactions of TASA and C＇I＇X or CAZ were ascertained by evaluating the fractional inhibitory concentration index （FICI）. Results： The antibacterial activity of either TASA itself or in combination with C＇IX or CAZ was found. The minimum inhibitory concentration （MICs） of TASA against the ESBLs producing isolates was 12.5 mg/mL. In the combinations with a sub-inhibitory concentration of TASA, a synergistic effect on CTX and CAZ against the ESBLs producing isolates was observed. Similarly, the isolates exposed to lower dose of TASA yielded an increased susceptibility to CTX and CAZ by 8-16 folds determined by microdilution assay. Moreover, enzymatic detection of ESBLs demonstrated that TASA induced reversal resistance to CTX and CAZ partially by a mechanism of inhibition of ESBLs activity in these isolates. Additionally, in the tested isolates following the exposure of TASA, molecular analysis verified the SHV-type beta-lactamase encoding ESBL gene in these isolates, and no mutation was introduced into the ESBL gene. Conclusions： These results suggest that TASA could be used as a source of natural compound with pharmacological activity of reversal resistance to antimicrobial agent. These findings also indicated that the application of the TASA in combination with antibiotics might prove useful in the control and treatment of infectious diseases caused by the ESBLs producing enterobacteriaceae.

Protective Effects and Mechanism of Total Coptis Alkaloids on Aβ_(25-35) Induced Learning and Memory Dysfunction in Rats

Objective： To observe the effect of total coptis alkaloids （TCA） on β -amyloid peptide （A β 25-35） induced learning and memory dysfunction in rats, and to explore its mechanism. Methods： Forty male Wistar rats were randomly divided into four groups： the control group, the model group, the TCA low dose （60 mg/kg） group and the TCA high dose （120 mg/kg） group, 10 in each. A β 25-35 （5 μl, 2 μg/μl） was injected into bilateral hippocampi of each rat to induce learning and memory dysfunction. TCA were administered through intragavage for consecutive 15 days. Morris Water Maze test was used to assess the impairment of learning and memory; concentration of malondialdehyde （MDA） in cerebral cortex was determined by thiobarbituric acid reactive substance to indicate the level of lipid peroxidation in brain tissues; activity of manganese-superoxide dismutase （Mn-SOD） in cerebral cortex was determined by xanthine-oxidase to indicate the activity of the enzyme; and NF-κB protein expression in cerebral cortex was measured by SP immunohistochemistry. Results： （1） Morris Water Maze test showed that, during the 4 consecutive days of acquisition trials, the rats in the model group took longer latency and searching distance than those in the control group （P〈0.01）, which could be shortened by high dose TCA （P〈0.05）; during the spatial probe trial on the fifth day, the rats in the model group took shorter searching time and distance on the previous flat area than those in the control group （P〈0.01）, which could be prolonged after TCA treatment （for low dose group, P〈0.05; for high dose group, P〈0.01）. （2） Analysis of cerebral cortical tissues showed that, compared with the control group, MDA level got significantly increased and Mn-SOD activity decreased in the model group （both P〈0.01）. After having been treated with TCA, the MDA level got significantly decreased （P〈0.05 and P〈0.01 respectively for low and high dose group）, while relative increase of Mn-SOD activity only appeared in high dose group （P〈0.05）. （3） Immunohistochemistry analysis showed the protein expression of NF- κB got significantly increased after modeling, while high dose TCA can significantly inhibit it. Conclusion： TCA could improve A β 25-35 induced dysfunction of learning and memory in rats, and its protective mechanism is associated with its actions in decreasing MDA level, increasing Mn-SOD activity and inhibiting the expression of NF-κB in cerebral cortex.

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.

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.

Effects of Total Alkaloids in Buxus microphylla Leaves on Aorta Smooth Muscle of Rats and Their Mechanisms

Objective To investigate the effects of total alkaloids in Buxus microphylla leaves(ABML)on isolated rats thoracic aorta rings,and then to explore the possible mechanisms underlying the effects.Methods Thoracic aortas of Wistar rats were isolated,removed,and mounted onto an organ bath.The effects of ABML at different concentration on the contraction of isolated thoracic aorta rings(with and without endothelium)precontracted with KCl or PE were observed with organ bath technique.Dose-effect curves of CaCl2 were recorded by organ bath technique.The concentration of intracellular Ca 2+ ([Ca 2+ ]i)increased by PE,KCI,and caffeine in the presence of ABML was determined using Ca 2+ sensitive fluorescence indicator Fura-2/AM loaded thoracic aorta vascular smooth muscle (VSM)cells of rats.Results In aorta rings precontracted with PE and KCl,ABML produced concentration- dependent relaxation in both intact and denuded endothelium ring groups.There was no difference in the inhibition of contraction between the intact and denuded endothelium ring groups at the same concentration.Exposure of isolated thoracic aorta rings to ABML led to a significant reduction in the contracting response induced by CaCl2,and shifted the cumulative concentration-response curves to right.ABML could significantly inhibit the extracellular Ca 2+ influx induced by PE and KCl under[Ca 2+ ]0 of 1.5 mmol/L,with inhibitory ratios of 40.2%and 49.9%,respectively.In the case of Ca 2+ -free,ABML could significantly inhibit the intracellular Ca 2+ release induced by PE,with inhibitory ratio of 72.4%.Conclusion ABML relaxes thoracic aorta VSM cells by suppressing influx of extracellular Ca 2+ via voltage-dependent Ca 2+ channel and receptor-operated Ca 2+ channel.

Research on Hypnotic and Anticonvulsant Activities of Total Alkaloids in Leaves of Eucommia ulmoides

Objective To study the hypnotic and anticonvulsant activities of the leaves of Eucommia ulmoides（LEU）,the total alkaloids were extracted from LEU using water-acid method.Methods Mice were divided into five groups,including groups contained total alkaloids in three different doses,negative and positive control groups.Direct hypnotic experiment in mice,pentobarbital sodium synergistic experiment,and anticonvulsant experiment were used.The numbers of spontaneous activities,sleep rate,sleep latency,sleep time,convulsion rate,and convulsion latency of the mice were recorded and analyzed.Results The total alkaloids from LEU could increase the sleep rate,significantly lengthen the sleep time,and shorten the sleep latency of mice.Even the low dose of total alkaloids（0.33 g/kg）showed significantly different activities with negative control group（physiological saline）with the synergistic effect of the superthreshold dose of pentobarbital sodium.Furthermore,the total alkaloids efficiently inhibited the convulsion caused by nikethamide.Conclusion The total alkaloids from LEU have the excellent sedative,hypnotic,and anticonvulsant activities in mice,with high safety and little drug side effects.Therefore,they have the protential development prospects in sedative-hypnotic drugs.

First total synthesis of isoquinolinone alkaloid marinamide and its methyl ester

The first total synthesis of isoquinolinone alkaloid marinamide 1 and its methyl ester 2 was described. The key steps involved a regioselective Frieclel-Crafts reaction of 1-benzyl-1H-pyrrole to form the intermediate 8.

青风藤总生物碱聚乳酸微球的制备

聚乳酸(PLA)及其共聚物具有良好的生物相容性和生物降解性,在人体内无积聚,最终可完全降解为二氧化碳和水。