ABSTRACT
ObjectiveTo explore the antitumor effect of solanine and its mechanisms.MethodsThe in vivo antitumor effect of solanine was observed using models developed through in vivo transplantation of tumor cells; In vitro lines of sensitive antitumor cells were selected from the digestive system using MTT assay; The effect of solanine on cell morphology was observed using transmission electronic microscopy; The morphology of apoptotic cells was observed using Annexin V/PI double staining and laser confocal scanning microscopy (LCSM); The rate of cell apoptosis was measured using Annexin V/PI double staining and flow cytometry; The concentration of intracellular Ca2+ ([Ca2+]1) was determined using Fluo-3/AM staining and LCSM; The membrane potential of cellular mitochondria was determined using TMRE staining and LCSM; The protein expression of Bcl-2 and Bax was measured using immunological marking and LCSM; And the activity of caspase-3 was measured using the colorimetric method.ResultsSolanine could inhibit the growth of tumor weight in S180 tumor-bearing mice and prolong the survival time of H22 tumor-bearing mice.MTT assay revealed that HepG2 cells were quite sensitive to solanine because solanine could induce morphological changes in HepG2 cells,with the rate of early apoptosis being 4％,8.5％,and 20.1％,for HepG2 cells treated for 24 h with solanine at concentration of 0.4,2,and 10 μg/mL,respectively.Solanine could raise the [Ca2+]i and lower the membrane potential.It could reduce the protein expression of Bcl-2 while increase that of Bax,thus increasing the activity of caspase-3.ConclusionThe obvious antitumor activity of sotanine in human hepatocarcinoma is demonstrated.This inhibitory effect is achieved through solanine decreasing the Bcl-2/Bax ratio,thus increasing [Ca2+]i,which could enhance the enzymatic activity of the caspase family,thus inducing the apoptosis of HepG2 cells.
ABSTRACT
<p><b>OBJECTIVE</b>To study how the way in which betaine promotes the proliferation of mouse spleen lymphocytes is related to calcium channels.</p><p><b>METHOD</b>BALB/c mice were used for this experiment. Mouse spleen lymphocytes were obtained through in vitro cultivation after they had been separated, and were divided into a negative control group, a Con A group, and 0.04, 0.4, 4, and 20 mmol x L(-1) betaine groups. MTT was used to observe the effect of betaine on the proliferation of mouse spleen lymphocytes; flow cytometry was used to measure the changes in the cell cycle of mouse spleen lymphocytes; and laser confocal scanning microscopy was used to observe the changes in the intracellular [Ca2+]i of mouse spleen lymphocytes after betaine or different calcium channel blockers were applied.</p><p><b>RESULT</b>Betaine was found to promote the proliferation of mouse spleen lymphocytes 12 h after it had been applied in vitro in concentrations of 4 and 20 mmol x L(-1). It was also found to promote the proliferation of mouse spleen lymphocytes 24 h and 48 h after it had been applied in vitro in concentrations of 0.04, 0.4, 4, and 20 mmol x L(-1), with the effect being most marked for the 4 mmol x L(-1) group 24 h after its application. It was found to facilitate the entry of mouse spleen lymphocytes from the G0/G1 to the S phase 4, 6, 18, and 24 h after it had been applied to mouse spleen lymphocytes in a concentration of 4 mmol x L(-1), with the effect being most marked at 18 h after its application. Intracellular [Ca2+]i in mouse spleen lymphocytes increased significantly (P < 0.01) 6, 12, 18 h after 4 mmol x L(-1) betaine had acted on the lymphocytes, with the effect being most marked at 6 h. The calcium channel blockers nifidipine, diltiazem, mibefradil, and genistein had no effect on the increase of the intracellular [Ca2+]i in mouse spleen lymphocytes due to the application of betaine, while verapamil, mycifradin, heparin, and procaine could block such increase.</p><p><b>CONCLUSION</b>Betaine facilitates the entry of mouse spleen lymphocytes from the G0/G1 into the S phase by raising the intracellular [Ca2+]i in these cells, thus promoting their proliferation. Intracellular [Ca2+]i increases mainly in two ways: (1) By affecting the alpha1 subunit of the L-type voltage-gated calcium channel with mediation by G proteins and thus leading to an efflux of intracellular calcium: (2) By affecting the IP3R and RyR calcium channels of the intracellular calcium stores and thus leading to the release of intracellular calcium.</p>
Subject(s)
Animals , Female , Male , Mice , Betaine , Pharmacology , Calcium , Metabolism , Calcium Channels , Metabolism , Cell Cycle , Cell Proliferation , Lymphocytes , Cell Biology , Metabolism , Mice, Inbred BALB CABSTRACT
BACKGROUND: It is needed in the treatment of yearly increased nervous and mental diseases to seek safe and effective sedatives and tranquilizers from natural drugs. It is found in the previous experiments that the total alkaloid of equisetum pratense (TAEP) has the satisfactory sedative and tranquitrzing actions.OBJECTIVE: To analyze the inhibitory effect of TAEP on the central nervous system.DESIGN: A randomized control study taking experiment animals as the observing objects.SETTING: The Post-doctor Scientific Research Station, Institute of Materia Medica, Harbin Shangye University.PARTICIPANTS: The experiment was carried out in the Department of Pharmacology, Heilongjiang University of Traditional Chinese Medicine between March 2002 and January 2003. The drug was TAEP, and 24 Wistar rats were used.METHODS: Twenty-four rats were randomly divided into 3 groups: TAEP group (60 mg/kg), reserpine group (30 mg/kg) and saline group (the same volume), all the rats were given intraperitoneal injections of the above drugs in corresponding dosages, and then the contents of monoamine neurotransmitters in brain were determined by high performance liquid chromatography (HPLC)-electrochemistry (HPLC-EC) and HPLC-ultraviolet (HPLC-UV) respectively.MAIN OUTCOME MEASURES: The contents of monoamine neurotranmitters of norepinephrine, adrenalin, dopamine and 5-serotonin (5-HA) and its metabolites of DHPR, 5-hydroxyindoleacetic acid (5-HIAA) and homovanillic acid in the striatum and marginal area of brain in rats were observed.RESULTS: There were 8 rats in each group both before and after the experiment without abnormalities or death. TAEP significantly lowered the contents of monoamine neurotranmitter but increased the contents of neutral and acidic metabolites of monoamine neurotranmitters in the striatum of rats. TAEP significantly decreased the monoamine neurotranmitters and significantly elevated the contents of monoamine metabolites of 5-HIAA and homovanillic acid, but insignificantly increased the content of DHPR in the marginal area of brain (P ＞ 0.05).CONCLUSION: The sedative and tranquilizing actions of TAEP are associated with the decreased contents of central monoamine neurotranmitters. TAEP has an action on the evacuation of monoamine which is similar to reserpine, which may be considered as one of the mechanisms of its sedative and tranquitrzing actions on central nerve system.
ABSTRACT
Object To study the inhibitory effect of Sargassum fusiforme (Harv.) Setch. polysaccharide (SFPS) on six different kinds of tissue tumor by antitumor experiments in vitro, and to observe tissue speciality of tumor treatment and reveal antitumor effect mechanism. Methods MTT and colony forming methods were adopted to study the antitumor activity of SFPS in vitro. Influences of SFPS on cell cycle and apoptosis were observed. LCM was adopted to measure [Ca 2+ ]i of the cells marked by Fluo 3/AM probe. Results SFPS had content antitumor activities to SGC 7901 and COLO 205. It prevented the transforming of SCG 7901 from G 0/G 1 period to S period. And it increased the apoptosis index (APO%). Applying SFPS, [Ca 2+ ]i increased first and then decreased. Further, after giving CaCl 2, [Ca 2+ ]i increased again. Conclusion By inducing the apoptosis of tumor cells, SFPS shows content antitumor activities to SGC 7901 and COLO 205. SFPS could increase the [Ca 2+ ]i in SGC 7901. And the Ca 2+ are from cellular storage.
ABSTRACT
Object To study the effects of total alkaloids in Equisetum pratense Ehrh. (TAEP) on the activity of monoamine oxidase B (MAO B) in mice brain, to reveal the mechanism of its inhibitory action on the central nervous system (CNS). Methods The activity of MAO B was determined by UV spectrophotometry. Results TAEP can markedly activate MAO B in cerebrum and antagonise the inhibition of MAO B activity by nialamide. Conclusion TAEP is a MAO B agonist with an action for the evacuation of monoamine, this may be considered as one of the mechanisms of TAEP sedative and tranquilizing action on CNS.
ABSTRACT
Object To study the antitumor mechanism of sea algae polysaccharide (SP). Methods The membrane fluidity of the S 180 and H 22 mice was observed by DPH fluorescent probe method. Results SP could increase the membrance fluidity of the S 180 and decrease that of the H 22 . Conclusion SP showed the antitumor effect through returning the tumor cell membrance fluidity to the normal one.
ABSTRACT
Object To study the effect of Haimidin(HMD) on erythrocyte membrane of H 22 ascites tumor bearing mice and DNA synthesis in human gastric tumor cell Methods By fluorescence spectrophotometry and laser scanning confocal microscopy Results HMD can lower erythrocyte microviscosity and elevate membrane lipid fluidity of H 22 ascite tumor bearing mice It also reduced DNA fluorescent intensity and DNA content of gastric tumor cells in vitro Conclusion HMD can improve blood circulation of tumor bearing mice, enhance immune adherence of erythrocyte on tumor cells and displays its antitumor activity by interferring DNA synthesis of tumor cells
ABSTRACT
Objective To explore the effects of humulon on kinetic parameters of N-acetyltransferase-1(NAT1) of human gastric cancer SGC-7901.Methods Employing HPLC,using para-aminobenzoic acid(PABA) as substrate,in intact SGC-7901 cells and their cytoplasm,taking the speed of PABA being acetylated to Ac-PABA by NAT1 as the rate of NAT1,using double reciprocal plot,taking the reciprocal of concentration of PABA and reaction rate of NAT1 as coordinates,regression equation was obtainied and the Michaelis constant(Km) and maximum reaction velocity(Vmax) were calculated.Results Study on enzyme kinetics demonstrated,as for intact SGC-7901 cells,Km and Vmax of control group were(3.910?0.087) ?mol/L and(0.306 0?0.006 7) pmol/L(1?106 cells),respectively,Km and Vmax of the humulon group were(3.830?0.123) ?mol/L and(0.275 0?0.005 8) pmol(1?106 cells),respectively.As for the cytoplasm of SGC-7901 cells,Km and Vmax of control group were(760.2?210.2) ?mol/L and(0.191 0?0.043 7) pmol/(mg?min),Km and Vmax of the humulon group were(449.0?72.9) ?mol/L and(0.094 0?0.010 4) pmol/(mg?min).Statistically,as for intact SGC-7901 cells or their cytoplasm,there was no difference of the Km between control group and humulon group,but there was remarkable difference of Vmax between control group and humulon group,P
ABSTRACT
Objective To study the anti-tumor effects of humulon on arylamine N-acetyltransferase-1(NAT1)activity.Methods Employing HPLC,using PABA as substrate,in intact SGC-7901 cells and their cytoplasm,making PABA being acetylated to Ac-PABA by NAT1 as the activity of NAT1.Reverse transcriptase polymerase chain reaction(RT-PCR)assay was used to study the expression of the NAT1 mRNA.Results The results show that humulon could inhibit the production of Ac-PABA in intact SGC-7901 cell and the cytoplasm,the production of Ac-PABA was gradually increased with the interaction time increasing.But comparing with corresponding negative control group's,the production of Ac-PABA was decreased evidently and the humulone could inhibit the expression of NAT1 mRNA.Conclusion Humulon could prevent the occurrence and deterioration of cancer.Its mechanisms can be attributed to its effect on decreasing the production of acetylation of carcinogenic aromatic amines,which is acetylated from aromatic amines,and inhibiting the NAT1 activity and expression of NAT1 mRNA.
ABSTRACT
Objective To study the relationship between apoptosis and mitochondria damage.Methods The cell morphology was observed under inverted microscope.Staining with Annexin V/PI,the cell apoptosis was observed by laser confocal scan microscope and the apoptosis rate was analyzed by flow cytomety. Transmission electron microscope(TEM) was used to observe mitochondrial structure.ROS was determined by confocal.Colorimetry assay was adopted to determine the reduced glutathione hormone (GSH).Results The cell number in solanine groups was fewer than that in control group,and some died.Staining with Annexin V/PI,Annexin V-FITC was high fluorescent in solanine groups,which is obviously apoptosis features.The early apoptosis rate is 4.0%,8.5%,and 20.1%inducing by 0.4,2,and 10 mol/L solanine for 24 h,respectively.Swelling mitochondria,absence of mitochondria crests and vesicles were found in mitochondria of HepG2 cell treated by solanine for 24 h.The ROS levels increased and GSH level decreased in solanine groups.Conclusion Solanine could induce the apoptosis and the effect may be attributed to decrease GSH,which interrupts the immediate remove of ROS so as to damage the mitochondria.
ABSTRACT
Object To reveal the mechanism of inhibitory action on the central nervous system (CNS) of total alkaloids of Equisetum pratense Ehrh. (TAEP). Methods Contents of monoamine neurotransmitters in brain of rats were determined by HPLC-EC and HPLC-UV. Results TAEP can significantly decrease the contents of monoamine neurotransmitter in the straite body and the bordering area of rat forebrain, increase the contents of neutral and acidic metabolites in the straite body, and increase the contents of 5-HIAA and HVA of the monoamines in the bordering area of rat forebrain. But its raise of DHPG showed no statistical significance. Conclusion TAEP has an evacuating action of monoamine similar to Reserpine. This may be considered as one of the mechanisms of sedative and tranquilizing effect CNS.
ABSTRACT
Object To study the effect of total alkaloids of Equisetum pratense Ehrh. (TAEP) on the contents of amino acid neurotransmitters and Ach in rat brain to reveal the mechanism of TAEP inhibitory action on the central nervous system (CNS). Methods Contents of amino acid neurotransmitters and Ach in rat brain were determined by double-wavelengh scan and GPI mensuration. Results TAEP could not influence four kinds of content of amino acid neurotransmitter (glutamic acid, glycin, ?-aminobutyric acid, aspartic acid), but TAEP could significantly lower the content of Ach in striatum of rat. Conclusion The inhibition of TAEP to CNS is attained by lowering the content of Ach in striatum to affect DA-2 receptor, and it is irrelevant to the amino acid neurotransmitters (glutamic acid, glycin, ?-aminobutyric acid, aspartic acid).
ABSTRACT
Objective To study the effect of Solanum nigrum alkaloid on erythrocyte immunity function. MethodsStudying the adherence between erythrocyte immunity and tumor-cell of S180 and H22 tumor-bearing mice; By determining with fluorescence polarization (P) and accounting the micro viscosity (?) with DPH as a probe the erythrocyte membrane fluidity of the S180 and H22 tumor-bearing mice could be investigated. ResultsS. nigrum alkaloid could increase the ratio of adherence anadem between erythrocyte and tumor-cell and promote the erythrocyte membrane fluidity of the two tumor-bearing mice. ConclusionThe inhibition of S. nigrum alkaloid on tumor can be accomplished through improving the erythrocyte membrane fluidity of two tumor-bearing mice and renewing erythrocyte immunity of tumor-bearing mice.
ABSTRACT
Objective To study the effect of Solanum nigrum total alkaloid(SNTA) on sialic acid(SA) and blocking degree of erythrocyte membrane in S_(180) tumor-bearing mice.Methods Using chromatometry to determine the erythrocyte membrane sialic acid and erythrocyte membrane blocking degree.Results SNTA could increase erythrocyte membrane sialic acid level and heighten erythrocyte membrane blocking degree in S_(180) tumor-bearing mice significantly(P
ABSTRACT
Objective To observe the effects of solanine on DNA and RNA level in tumor cell of S_(180) and H_(22) tumor-bearing mice.Methods S_(180) and H_(22) mice were divided into solanine(37.50、18.75,and(9.37) mg/kg) groups,negative control group and cytoxan(30 mg/kg) group,whom were given drugs by sc.Levels of RNA and DNA in tumor cell membrane in every groups were examined,respectively by laser scanning confocal microtechnic technology.Results Solanine(37.50、18.75 mg/kg) groups reduced the ratio of RNA and DNA in tumor cell of both S_(180) and H_(22) mice.Conclusion Solanine can reduce the ratio of RNA and DNA in tumor cell of S_(180) and H_(22) mice,which may be one of the mechanisms of solanine's(antitumor) effect.
ABSTRACT
Arylamine N-acetyltransferase(NATs)is one of phase Ⅱ drug-metabolizing enzymes,which is very important in drug metabolizing and detoxification.It may become a new target for studying pharmacological effects.The study on NATs is hitherto its crystal structure or polymorphisms among different species,different ethnicity,same species,and same ethnicity.However,the study on its physiological and pharmacological effects is unwontedly reported.In the paper the research work on NATs is summarized from the following aspects:the structure,carcinogenic mechanism,endogenous activities,inhibitor,substitute products,metabolic kinetics,polymorphism and relation with diseases,and detected methods of NATs.