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AIM To investigate the inhibitory effects on the growth of SMMC-7721 human hepatocellular carcinoma cell line and its mechanism by theasinesin. METHODS MTT assay,colony formation test, 3H-TdR, 3H-Leu incorporation test, determination of the intracellular cAMP and cGMP level, in situ hybridization were used. RESULTS Theasinesin at 50 mg*L-1 could significantly inhibit the growth and colony formation of SMMC-7721 cells. Theasinesin could strongly inhibit DNA and protein synthesis and increase intracellular cAMP levels. C-myc gene expression was significantly decreased and p53 gene increased strongly in a dose-dependent manner.CONCLUSION Theasinesin demonstrates significant inhibitory effects on the growth of SMMC-7721 cells. Its mechanisms may involve the inhibition of DNA and protein synthesis ,the elevation of intracellular cAMP levels, inhibition of c-myc gene expression and enhancement of p53 gene expression.
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OBJECTIVE:To study the antitumor activity and immunomodulatory effects of theasinesin on mice transplanted tumor and tumor-bearing mice.METHODS: With three mice transplanted tumor models i.e. Ehrlich ascites tumor, sarcoma 180 and hepatic carcinoma H22,we strdied the antitumor activity of theasinesin .The immunomodulatory effects of theasinesin on S180-bearing mice were measured by delayed type hypersensitivity(DTH), cleaning charcoal particles method, and splenocyte proliferation test .RESULTS: Theasinesin at doses of 400,200, 50mg/kg could markedly inhibite the growth of Ehrlich ascites tumor(solid tumor) .Although theasinesin didn't have significant inhibitory effects on sarcoma 180 and hepatic carcinoma H22 in routine oral administration, the average inhibitory rates of each group, in manner of preventive administration, were all greater than 30% .Theasinesin at doses of 50,100,200mg/kg could significantly restore the decreased DTH in S180-bearing mice. It could also markedly increase K, a and promote proliferation of activated T cells in S180- bearing mice. CONCLUSION :Theasinesin has significant inhibitory effects on mice transplanted tumors. It can also enhance the decreased immunological function on tumor-bearing mice.
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AIM: To study the effects of theasinesin(TS) on proliferation and apoptosis of murine lymphocytes. METHODS: MTT method was employed to investigate the proliferatory response of murine splenocytes. Cellular morphological observation, DNA agarose gel electrophoresis and flow cytometry analysis were used to study the effects of TS on apoptosis of murine thymocyte and splenocyte. RESULTS: 50,150 and 500 mg?L -1 TS could markedly promote ConA-induced splenocyte proliferation in mice. In the cellular morphologic observation and DNA biochemical analysis, murine thymocytes cultured for 20 h in vitro showed keryopyk-nosis, condensation of cell, typical "ladder" on gel electrophoresis and typical apoptotic peak (AP) by FACS analysis. After treatment with 50, 150 and 500 mg?L -1 TS , DNA ladder and apoptotic peak were decreased gradually and apoptotic rates were reduced to 12.14%, 9.49%, 6.71% respectively from 19.87% in control group. But the apoptosis of murine splenocytes induced by 10 -5 mol?L -1 DEX was not affected by TS. CONCLUSION: TS can significantly promote splenocyte proliferation in mice and has an inhibitory effect on spontaneous apoptosis of murine thymocytes.
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Object To study the immunomodulatory effects of theasinesin (TS) on human peripheral blood mononuclear cells (PBMC) in vitro. Methods The proliferation response of lymphocytes and IL 2 production of human PBMC were detected by MTT method. IL 2R expression was determined by immunocytochemistry and indirect immunofluorescence. The intracellular cAMP level was assayed by radioimmunoassay. Results 50, 150, 500 ?g/mL TS could enhance phytagglutinin (PHA) induced human PBMC proliferation in a dose dependent manner. The IL 2 production and IL 2R expression of PHA activated PBMC were increased markedly under the presence of TS. TS also decreased cAMP level of PBMC and intensively weaken the cAMP upregulating effect of prostaglandin E 1 (PGE 1). Conclusion TS possesses a potential immunoenhancing effect which might be made by decreasing the cAMP level of PBMC, promoting IL 2 production and IL 2R expression.
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Objective To investigate the scavenging abilities of ginkgo albumin to reactive oxygen in vitro, as well as the protection and mechanism of ginkgo albumin on damaged DNA. Methods Lotus Seedpod Procyanidin (LSPC) and bovine serum albumin (BSA) were used as control sample, the effect of ginkgo albumin on removal of superoxide anion was determined by Pyrogallol-Luminol system. Scavenging ability of ginkgo albumin on hydroxide free radical was determined by CuSO4-Phen-Vc-H2O2, FeSO4-Luminol-H2O2, and FeSO4-Luminol systems. Luminol-H2O2 system was used to measure the scavenging effect on hydrogen peroxide. Preventive effect of ginkgo albumin on in vitro damaged DNA was determined by CuSO4-Phen-Vc-H2O2-DNA system. Results Ginkgo albumin possessed a good scavenging potency on reactive oxygen and protection on damaged DNA, but promoted oxidation in the FeSO4-Luminol-H2O2 and Luminol-H2O2 chemiluminescence systems. Conclusion Not every chemiluminescences system is suitable for investigating the antioxidant activity of ginkgo albumin.
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AIM To investigate the inhibitory effects on the growth of SMMC 7721 human hepatocellular carcinoma cell line and its mechanism by theasinesin. METHODS MTT assay,colony formation test, 3H TdR, 3H Leu incorporation test, determination of the intracellular cAMP and cGMP level, in situ hybridization were used. RESULTS Theasinesin at 50 mg?L -1 could significantly inhibit the growth and colony formation of SMMC 7721 cells. Theasinesin could strongly inhibit DNA and protein synthesis and increase intracellular cAMP levels. C myc gene expression was significantly decreased and p53 gene increased strongly in a dose dependent manner. CONCLUSION Theasinesin demonstrates significant inhibitory effects on the growth of SMMC 7721 cells. Its mechanisms may involve the inhibition of DNA and protein synthesis ,the elevation of intracellular cAMP levels, inhibition of c myc gene expression and enhancement of p53 gene expression.
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The progress is reviewed on the studies of procyanicdins biological and pharmacological activities in the past ten years, including antioxidant,protecting cardiovascular system,regulating immunocompetence, antiviral, anticancer, antiulcer and antidepressant,antimutagenic properties, etc.
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AIM To study the effects of procyanidins' extract from the lotus' seedpod(LSPC) on experimental myocardia injury in rats. METHODS Compared with diltiazem(10 mg?kg -1 ?d -1 ) and complex salvia (8 mg?kg -1 ?d -1 ), after one hour supplementation of LSPC, the model of myocardial injury induced by subcutaneous injection of isoproterenol (40 mg?kg -1 , twice )and by cardiac ischemia in situ were studied with rat hearts. RESULTS LSPC pretreatment (100 and 200 mg?kg -1 ) significantly antagonized the high serum of CK,CK MB,LDH, calcium and the high content of myocardial water, calcium, provided significant reduction in myocardial infarct size, increased the ratio between superoxide dismutase activity and malondialdehyde production in serum, lighted the degree of myocardial pathological damage. CONCLUSION LSPC provided a protection on ischemic myocardium in rats and the action is considered to be related to the inhibition of lipid peroxidation or myocardial Ca 2+ accumulation.
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AIM To investigate the mechanism of the anticancer effect of theasinesin. METHODS MTT assay, electronic microscopy technique, DNA gel electrophoresis, flow cytometric analysis with Annexin V Fluos staining,RT PCR and immunocytochemistry were performed for apoptosis and its mechanism. RESULTS After HL 60 cells were treated with theasinesin, HL 60 cells were characterized by condensation of the nuclear chromatin, margination against the nuclear envelope, and apoptotic body formation with scanning and transmission electronic microscopy. A typical DNA ladder was detected by agarose gel electrophoresis of DNA extracted from HL 60 cells. Flow cytometric analysis of HL 60 cells by Annexin V Fluos and propidium iodide double staining demonstrated that theasinesin induced apoptosis had dose effect and time effect relationships, but treatment with high dose or long time would induce cell necrosis principally. RT PCR and immunocytochemistry indicated that bcl 2 gene was reduced strongly in HL 60 cells. CONCLUSION\ Theasinesin induces apoptosis in HL 60 cells. The mechanisms for antitumor activity of theasinesin may involve in induction of apoptosis in cancer cells.
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Objective: To study the effects of procyanidins extract from the lotus seedpod (LSPC) on active oxygen radicals and lipid peroxidation. Methods: (1) The scavenging effects of LSPC on O ?- 2 and ?OH was investigated by using chemiluminescence method; (2) The effects of different concentrations of LSPC on RBC auto oxidation and LPO formation of rat liver mitochondria in vitro were observed; (3) The LPO level of liver homogenates incubated with Fe 2+ and H 2O 2 in vitro were determined; (4) the mice were treated with a single intragastric feeding of 5% CCl 4 soya bean oil solution (10ml/kg bw) after seven days supplementation of LSPC 100, 200, 400 mg/(kg bw?d). The activities of SOD, GST and the level of LPO in liver and plasma of CCl 4 toxic mice were determined. Results: In vitro, LSPC 169.0mg/L or 105.3mg/L was shown to markedly scavenge O ?- 2 produced in xanthine/xanthine oxidase and ?OH in Fenton systems respectively, and could significantly inhibit the formation of liver LPO. In vivo, LSPC (100,200mg/kg) evidently reduced the level of LPO and increase the activities of SOD and GST. Conclusion: It suggests that LSPC may be used as an antioxidant and might effectively protect liver from the injury due to lipid peroxidation.
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Objective:To study the effects of LSPC on impairment of learning and memory in mice induced by scopolamine and alcohol,and its possible mechanism.Method:One hundred male Kun-Ming mice were randomly divided into 2 batches,50 in each.Each batch was then divided into 5 groups(n=10):control,model,LSPC of 22.5,45,90 mg/kg bw.Group 3-5 was treated with lotus seedpod procyanidins(LSPC,dissolved in normal saline,22.5,45,90 mg/kg bw)by ig.each day for 20 d,while group 1-2 was treated with ig.normal saline.The model of learning and memory impairment was established by ip.3 mg/kg bw scopolamine hydrobromide or ig.40% alcohol in 0.1 ml/10 g bw respectively.Then,effect of LSPC on learning and memory in mice were tested by Y-maze.As to the mice whose learning and memory ability was impaired by alcohol,their brains were dissected out after Y-maze experiment for detecting the level of MDA,the activity of SOD and MAO-B.Results:The learning and memory abilities of mice impaired by scopolamine were remarkably improved after22.5 and 90mg/kg bw LSPC treatment.In the alcohol experiment,the impairment was significantly improved after three different doses of LSPC administration.The level of MDA was reduced,the activity of SOD increased and the activity of MAO-B decreased in mice that learning and memory was impaired by alcohol.Conclusion:LSPC can improve learning and memory abilities of the mice impaired by alcohol and scopolamine,and the mechanism in alcohol experiment probably lay on its suppressive effect on MAO-B activity and protection of neurons from oxidative stress.
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Objective:To study the anti-tumor activity of ginkgo albumin (GA) and its mechanism. Method: Molecular weight of GAⅡa was detected by MS (mass spectrum). The antioxidative effect of GA was studied by deoxyribonucleic acid method and nitroblue tetrazolium (NBT) photo-reduction, and the anti-tumor activity of GA on S180 sarcoma was studied both in vivo and in vitro. Results: Molecular weight of GAⅡa was 29 248u. GA had strong scavenging effect on superoxide radical and hydroxyl radical. The growth of S180 both in vitro and in vivo was inhibited significantly by GA. Conclusion: The anti-tumor activity of GA was probably related with its antioxidative effect.
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Objective: To study the contents and constituents of LSPC (procyanidins from Lotus Seedpod) and its chemoprophylactic effects on 9,10-dimethyl-1,2-benzanthracene (DMBA)-induced golden hamsters buccal-pouch carcinomas. Methods: ESI-MS was used to analyze LSPC, and the change of body weight, mortality during test, and the value of serum MDA, GSH-Px, T-SOD,the gross and pathological change of buccal-pouch mucosa were investigated when golden hamsters were via gastric intubation or the buccal pouch mucosa was smeared with 100mg/(kg bw?d)LSPC. Results and conclusion: The contents of LSPC exceeded 98% and mono-, di-, tri-, tetrameric procyanidins as well as di-, trimeric galic ester were constituted of LSPC with molecular weight ranging from 290-1154. LSPC had chemoprophylactic effects on DMBA-induced golden hamsters buccal-pouch carcinomas, and the effect was superior through smearing LSPC rather then via gastric intubation .
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Objective: To probe into the antioxidation mechanism of lotus seedpod in vivo.Methods:Nineteen 4-month old rats were randomly divided into control group and procyanidins group. After feeding 100 mg/kg LSPC for 35 d, MDA and SOD in skin and serum, GSH-Px and Hyp in blood and skin were measured.Results: MDA levels in skin and serum in the procyanidins group were obviously lower than that of the control(P
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Objective: To analyze and identify the carotenoid in gingko seed.Methods: The chemical composition of the pigment from gingko seed was identified as carotenoid by UV absorption spectrum. Its content was determined by spectrophotometric method and its composition was analyzed and identified by thin-layer chromatography and HPLC method.Results: Gingko contained carotenoids, roughly about 489 ?g/100 g in which lutein, ?-carotene and ?-carotene amounted to 69.20%,15.80% and 7.45% respectively. Conclusion: Gingko contained large quantity of carotenoids, mainly lutein, and next in order ?-carotene and ?-carotene.