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.

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.

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.

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.

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.

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).

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.

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.

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.