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Objective To observe the effects of diacetyldianhydrogalactito (DADAG) on the proliferation and apopotosis in lung cancer cell NCI-H460.Methods MTT assay was performed to determine the half inhibitory concentration of DADAG(2.88,5.75,11.50,23.00 and 46.00 μg/mL)for the NCI-H460 cells and colony formation assay was used to detect the ability of cell proliferation; through AO/EB staining method, morphological changes of apoptotic cell under fluorescent microscope were observed;cell apoptosis was detected by flow cytometry to test the effects of DADAG on NCI-H460;RT-PCR was used to detect the effect of DADAG on Bcl-2 and Bax mRNA expres-sion levels within the cell. Results Compared with that in blank control group, cell proliferation was inhibited in the group treated with DADAG; the number of colony formation decreased and AO/EB staining results showed that cell apoptosis was characterized by typical morphological changes such as swelling, shrinking and fragmentation. Flow cytometry detection results showed that apoptosis rate was increased in the group treated with DADAG(all P<0.05).RT-PCR results indicated that the expression of pro-apoptosis gene Bax was up-regulated,while expression of anti-apoptosis gene Bcl-2 down-regulated. Conclusions DADAG exhibits the inhibitory activity in lung cancer cell NCI-H460 and can induce the apoptosis.The potential mechanism maybe associated with up-regulating the ex-pression of Bax but down-regulating the expression of Bcl-2.
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Objective To study whether crystal-8 exerts analgesic effect on the central nervous system,and to observe the relationship between crystal-8 and receptors. Methods A total of 32 SD rats were randomly divided into the 0.9% sodium chloride solution group, positive control group ( rotundine 2 mg·kg-1 ) , high dose of crystal-8 group ( 2 mg·kg-1 ) and low dose of crystal-8 group ( 1 mg · kg-1 ) . The changes of pain threshold were measured by the thermal stimulation test following intracerebroventricular (ICV) injection.The SD rats or mice were randomly divided into the 0.9% sodium chloride solution group, receptor tool medicine group ( including naloxone, reserpine, haloperidol, scopolamine) , crystal-8 group, receptor tool medicine plus crystal-8 group. The pain threshold was detected by the thermal stimulation test at 15, 30, 45, 60, 90, 120 min after dosing, then the influence of analgesic effect of crystal-8 on the neurotransmitter was observed. Results Compared with the 0.9% sodium chloride solution group, the pain threshold of rats was improved after taking the crystal-8 by intracerebroventricular (ICV) injection(P<0.01).Compared with the crystal-8 group, the naloxone plus crystal-8 group, the haloperidol plus crystal-8 group, the scopolamine plus crystal-8 group couldn't increase the pain threshold, there was no significant difference among these groups(P>0.05).However, Reserpine plus crystal-8 group could significantly decrease the pain threshold in rats compared with the crystal-8 group(P<0.01).The analgesic effect of crystal-8 was interfered by reserpine. Conclusion The analgesic effect of crystal-8 may be involved in the central mechanism,which relates to monoamine neurotransmitters, but has nothing to do with the opioid receptor, M receptor, and the inhibition of central DA receptor.
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Objective To study whether crystal-8 exerts analgesic effect on the central nervous system,and to observe the relationship between crystal-8 and receptors. Methods A total of 32 SD rats were randomly divided into the 0.9% sodium chloride solution group, positive control group ( rotundine 2 mg·kg-1 ) , high dose of crystal-8 group ( 2 mg·kg-1 ) and low dose of crystal-8 group ( 1 mg · kg-1 ) . The changes of pain threshold were measured by the thermal stimulation test following intracerebroventricular (ICV) injection.The SD rats or mice were randomly divided into the 0.9% sodium chloride solution group, receptor tool medicine group ( including naloxone, reserpine, haloperidol, scopolamine) , crystal-8 group, receptor tool medicine plus crystal-8 group. The pain threshold was detected by the thermal stimulation test at 15, 30, 45, 60, 90, 120 min after dosing, then the influence of analgesic effect of crystal-8 on the neurotransmitter was observed. Results Compared with the 0.9% sodium chloride solution group, the pain threshold of rats was improved after taking the crystal-8 by intracerebroventricular (ICV) injection(P<0.01).Compared with the crystal-8 group, the naloxone plus crystal-8 group, the haloperidol plus crystal-8 group, the scopolamine plus crystal-8 group couldn't increase the pain threshold, there was no significant difference among these groups(P>0.05).However, Reserpine plus crystal-8 group could significantly decrease the pain threshold in rats compared with the crystal-8 group(P<0.01).The analgesic effect of crystal-8 was interfered by reserpine. Conclusion The analgesic effect of crystal-8 may be involved in the central mechanism,which relates to monoamine neurotransmitters, but has nothing to do with the opioid receptor, M receptor, and the inhibition of central DA receptor.
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Objective:To discuss the effects of panax notoginseng saponins (PNS) enteric-coated pellets on hemorrheology in rabbits.Methods:The rabbits were divided into the normal control group,the model control group,Xueshuangtong injection (lyophilization) group(15 mg·kg-1·d-1 ,im),PNS enteric-coated pellets groups respectively at high(45 mg·kg-1·d-1,ig),medium(30 mg·kg-1·d-1,ig) and low (15 mg·kg-1·d-1,ig) dose.The model was established by intragastric administration of high-fat diet.The whole-blood viscosity,plasma viscosity,erythrocyte aggregational index,crythrocyte index of rigidity and erythrocyte electro-phoresis rate in the groups were detected using hemorheological methods.Results:The above indices of hemorheology in the model control group were all significantly higher than those in the normal control group (P0.05).Compared with Xueshuangtong injection (lyophilization) group,PNS enteric-coated pellets group at medium dose could significantly reduce the whole blood middle shear viscosity(P<0.05).Conclusion:PNS enteric-coated pellets can reduce the whole-blood viscosity,plasma viscosity,erythrocyte aggregational index,crythrocyte index of rigidity and erythrocyte electro-phoresis rate,and effectively promote blood circulation and remove stasis,inhibit thrombosis formation and increase blood supply for heart and cerebral vessels.
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Objective To compare the immunoregulatory effects of Radix Millettia Speciosa (RM Speciosa) and Radix Millettia Championi (RM Championi)on immunosuppressed mice.Methods Kunming mice were randomly divided into normal control group,CTX model group,LMS positive group,RM Speciosa groups and RM Championi groups (20,10 and 5 g/kg).The mice were treated respectively with drug or NS once a day for consecutive 20 days.Mice in the other groups were injected intraperitoneally with CTX at days 8,10 and 12 to establish immunosuppressed mice model except the normal group.The changes of body weight,immune organ weight,white blood cell (WBC)number,carbon particle clearance capability of macrophages and delayed type hypersensitivity (DTH)of mice in all groups were determined and compared.Results Compared with that in CTX group,the WBC number was significantly increased (P0.05).Conclusion Both RM Speciosa and RM Championi can improve the immune function of CTX-induced immunosuppressed mice,and RM Speciosa is slightly superior to RM Championi in improving specific cellular immunity.
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Aim To evaluate the antitumor activity of dianhydrogalactitol ( DAG) in vitro, and further clarify its underlying mechanisms. Methods The inhibitory effect of DAG in NCI-H460 cells was detected by CCK-8 assay and colony formation assay. The morphology of cells treated with DAG was observed under optical mi-croscope. Nuclear morphology was captured by fluores-cence microscopy after Hoechst 33342 staining. Real-time PCR was used to analyze the expression level of topoisomerase Ⅱ ( Topo Ⅱ) mRNA. The protein ex-pression level of Topo Ⅱ was detected by Western blot. Additionally, molecular docking approaches were used to predict the interaction between DAG and TopoⅡ. Results DAG exhibited potent antitumor activity in NCI-H460 cells, and inhibited cell proliferation per-sistently. DAG obviously induced nuclear morphologi-cal changes of NCI-H460 cells. Furthermore, DAG could down-regulate the mRNA and protein expression level of Topo Ⅱ detected by Real-time PCR analysis and Western blot, respectively. Molecular docking predicted that DAG could bind to Topo Ⅱ. Conclu-sion DAG can significantly inhibit the proliferation of NCI-H460 cells, and its underlying mechanisms may involve the down-regulation of Topo Ⅱ mRNA and di-rect binding to Topo Ⅱ, leading to cancer cell death.
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BACKGROUND:Lymph-targeted tracing and therapy based on carbon nanotubes have been one of the hottest researches on targeting tumor diagnosis and treatment. To evaluate the accumulation of carbon nanotubes in axil ary lymph node can provide experimental evidences for developing nano-tracers and drug carriers which are more lymph-specific and more biocompatible. OBJECTIVE:To study the accumulation of the intravenously injected carboxylated single-wal ed carbon nanotubes in axil ary lymph nodes of Sprague-Dawley rats, and to evaluate their effect on blood cel s. METHODS:Sixty-four Sprague-Dawley rats were randomly divided into two groups. Rats in testing group were injected with carboxylated single-wal ed carbon nanotubes suspension (2 mg/kg), while those in control group were injected with 5%glucose solution (1 mL/kg), both through the tail vein, three times per week. Four periods of 7, 60, 90 and 120 days were set (the 120-day period referred to 90 days of administration fol owed by 30 days of drug withdrawal). At the end of each period, eight rats from each group were randomly picked out, to col ect blood samples via the abdominal aorta for blood routine test. Final y the axil ary lymph nodes were observed, and the lymph node samples of rats in the testing group were col ected and analyzed at 120 days by transmission electron microscope. RESULTS AND CONCLUSION:Compared with the control group, black staining of axil ary lymph nodes of rats in testing group was not obvious at the end of the 7-day period. However, with the increase of the dosing periods, the lymph nodes of the rats in the testing group became enlarged, firm and black stained, coupled with a significant rising in the percentage of blood neutrophils. After 30 days of drug withdrawal, the size of the rat axil ary lymph nodes was reduced and black staining partly faded, with the decreasing of blood neutrophil percentage. Under the transmission electron microscope, abundant carboxylated single-wal ed carbon nanotubes were uptaken by lymphocytes to form a large number of phagocytic vacuoles after drug withdrawal for 30 days. It indicates that the short-term tracing of rat axil ary lymph nodes by carboxylated single-wal ed carbon nanotubes injected through the tail vein is relatively weak, while the long-term intravenous injection can cause their accumulation in rat axil ary lymph nodes, coupled with the increase of neutrophils;after drug withdrawal, the carboxylated single-wal ed carbon nanotubes can be slowly cleared by the lymph nodes.
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Objective To establish a HPLC method for determination of naringenin and apigenin in Premna fulva. Methods The SHISEIDO￣SPOLAR C18(250 mm×4.6 mm,5μm) was used as analytical column.The mobile phase consisted of methanol￣0.2% phosphoric acid (42:58) with isocratic elution at a flow rate of 1.0 mL.min-1 .The detection wavelength of naringenin and apigenin was 288 nm and 340 nm, respectively.Column temperature was set at 35 ℃ , the injection Volume was 10 μL. Results Naringenin and apigenin had a good linear relationship in the concentration range of 0.180 ~ 3.60 μg (r =0.999 9) and 0. 0052 ~ 0. 1040 μg ( r = 0. 999 9), respectively. Conclusion The method is accurate and reliable. It is appropriate for the quantitative determination of naringenin and apigenin in Premna fulva and its preparations.
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<p><b>OBJECTIVE</b>To establish the characteristic fingerprint of Zanthoxylum nitidum by HPLC, and to provide a reference for the quality control of Z. nitidum in the market.</p><p><b>METHOD</b>The established HPLC characteristic fingerprint of Z. nitidum, combined with similarity evaluation and system clustering analysis method, were applied to distinguish 25 batches of samples purchased from market preliminarily, to identify the authenticity and quality of Z. nitidum ingredients.</p><p><b>RESULT</b>In the 25 batches of samples purchased from market, only 8 batches were identified as genuine with good quality, 7 batches were identified as defective, 7 batches were identified as common counterfeit Toddalia asiatica, and 3 batches were identified as counterfeit.</p><p><b>CONCLUSION</b>This method is accurate, convenient and reliable. It can be used for identification and quality control of Z. nitidum ingredients.</p>
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Chromatography , Drugs, Chinese Herbal , Chemistry , Reference Standards , Quality Control , Zanthoxylum , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To develop a method for elucidating " chromatography-efficacy" relation of the extract of Zanthoxylum nitidum on the gastric cancer cells.</p><p><b>METHOD</b>After obtaining the tumor inhibition rate and fingerprint peak data through MTT and HPLC, "chromatography-efficacy" relation was established by an appropriate statistical method.</p><p><b>RESULT</b>The gastric cancer "chromatography-efficacy" relation of Z. nitidum was established by step-back technique.</p><p><b>CONCLUSION</b>The "chromatography-efficacy" relation has statistically significant and practical significance, so it has reference value in some way.</p>
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Humans , Antineoplastic Agents, Phytogenic , Pharmacology , Cell Line, Tumor , Cell Proliferation , Chromatography, High Pressure Liquid , Methods , Drugs, Chinese Herbal , Pharmacology , Stomach Neoplasms , Drug Therapy , Zanthoxylum , ChemistryABSTRACT
Aim To investigate the anti-tumor effect of nitidine chloride(NC)on human HepG2 hepatocellular transplanted tumor in nude mice and its effect on topoisomerase.Methods The subcutaneous transplantable tumor model of human liver cancer in nude mice was established and the anti-tumor effect of NC was calculated.The effects of NC on TopoⅠ/Ⅱ mediated-pBR322 DNA relaxation were measured by using agarose gel electrophoresis.Results NC inhibited significantly the growth of hepatoma,The inhibitory rate at the dose of 2.5,5,10 mg·kg~(-1) was 12.06%,35.63% and 60.91% respectively.At the concentration of 6.25 μmol·L~(-1),NC completely inhibited the pBR322 DNA cleavage mediated by TopoⅠ;at the concentration of 25 μmol·L~(-1),NC completely inhibited the pBR322 DNA cleavage mediated by Topo Ⅱ.Conclusion Nitidine Chloride can inhibit hepatic carcinoma growth in nude mice,The anti-tumor mechanism is probably related to the inhibitory effect on Topo.
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<p><b>OBJECTIVE</b>A high performance liquid chromatography (HPLC) method was developed to determine the concentration of nitidine chloride in plasma and successfully applied to study pharmacokinetics after i.v. administration in rabbits.</p><p><b>METHOD</b>Twelve rabbits, randomized into 2 groups , were given i.v. at the dose of 4, 6 mg x kg(-1) respectively. Chloramphenicol was used as an internal standard. Nitidine chloride was extracted from plasma with ion pair reagent, and was determined by HPLC.</p><p><b>RESULT</b>The calibration curves of nitidine chloride was linear in the range of 0.03-2.04 mg x L(-1). Its recoveries were more than 95%, intra-day and inter-day precisions were lower than 6%. The concentration-time curve of nitidine chloride in rabbits after i.v. of 4 and 6 mg x kg(-1) were shown to fit a two-compartment model, the main pharmacokinetic parameters showed no significant difference between the low and high dosage, and the AUC values are directly relative to doses. T1/2alpha were (5.46 +/- 0.89), (4.76 +/- 0.33) min respectively, T1/2beta were (263.33 +/- 16.4), (274.71 +/- 16.52) min respectively, AUC were (46.56 +/- 1.80), (69.19 +/- 2.30) microg x min(-1) x mL(-1) respectively.</p><p><b>CONCLUSION</b>It is first time to establish the HPLC method to determine the concentration of nitidine chloride in rabbits plasma. The method is sensitive, accurate and reproducible. It is first time to study the pharmacokinetic characters of nitidine chloride in rabbits after i.v. administration, the elimination of nitidine chloride is linear pharmacokinetics.</p>
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Animals , Female , Male , Rabbits , Benzophenanthridines , Blood , Pharmacokinetics , Chromatography, High Pressure Liquid , Methods , Drugs, Chinese Herbal , Pharmacokinetics , Random AllocationABSTRACT
OBJECTIVE: To establish an HPLC-UV method for the content determination of noradrenaline (NA) in plasma of spontaneous hypertension rats (SHR).METHODS: The determination was performed on C18 column with mobile phase consisted of methanol-0.15 mol?L-1 potassium dihydrogen phosphate buffer (1 ∶ 9,V/V,pH 6.0) at flow rate of 1.0 mL?min-1.The detection wavelength was set at 280 nm and column temperature was room temperature.RESULTS: The linear range of NA was 0.5～40 ?g?L-1 (r=0.999 9) with an average recovery of 97.49% (RSD
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Aim To investigate the apoptosis-inducing effects of AgLA2 on lung carcinoma cells SPC-A-1 and its mechanism in vitro.Methods The MTT assay was used to assess the proliferation of SPC-A-1 cells treated with AgLA2 in vitro.Apoptosis-inducing effects was investigated by DNA agarose gel electrophoresis,cell morphology and Elisa.RT-PCR was used to measure the expression of bcl-2 and bax mRNA,and immunocytochemistry was used to measure the expression of bcl-2 and bax protein.Results The IC50 of AgLA2 to SPC-A-1 cells was(3.447?0.436)mg?L-1.Treated with AgLA2,typical nuclear chromatine condensation and fragmentation were observed.The concentration of Caspase-3 in the group treated with AgLA2 was higher than that of the control group.Treated with AgLA2,bcl-2 mRNA,protein expression decreased while bax mRNA,protein expression increased.Conclusions AgLA2 can inhibit proliferation and induce apoptosis of SPC-A-1 cells.Its mechanism of action may be related to changing the ratio of bax/bcl-2 and the set-point of apoptosis,making the apoptosis power hold dominance.
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Aim To study protective effects of aFGF on rats' renal tubular epithelial cells injured by gentamicin in vitro.Methods After renal cortice were grounded,filtered and digested with 0.25%trypsin,renal tubular epithelial cells were cultured.The injury models of epithelial cells of renal tubules were established by gentamicin and then the protective roles of aFGF in injured renal tubular epithelial cells were observed.Results ① By observing the morphology of the epithelial cells and comparing GM group with control group,CAT、SOD、Na~+,K~+-ATP enzyme activities decreased,while NAG activity and MDA level increased(P
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Aim To study the anti-inflammatory effect of Yinhuang micro-enema compound (YHMEC) in vitro and its possible anti-inflammatory mechanisms. Methods Peritoneal macrophages (PM?) obtained from Wistar rats were randomly divided into four groups: control group, lipopolysaccharide (LPS)-induced group, YHMEC intervention group and dexamethasone intervention group. The morphological changes of cells were observed under convert microscope. Peritoneal macrophage viability was test with MTT. The levels of tumor necrosis factor?(TNF-?) and interleukin-6 (IL-6) were measured using ELISA. The translocation of nuclear factor-kappa B (NF-?B) p65 was detected with immunocytochemical method. Results The levels of TNF-? and IL-6 were increased significantly when PM? were induced by LPS and p65 were translocated from the cell cytoplasm into the nucleus. TNF-?,IL-6 secretions and translocation of NF-?B induced by LPS were inhibited by YHMEC. Conclusion The anti-inflammatory effects of YHMEC may act at least partly through inhititing the translocation of NF-?B and thus depress the expressions of TNF-? and IL-6.
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AIM: To detect the inhibition effect of nitidine chloride on proliferation of human liver cell L-02 and human kidney cell 293 and the protective effect of acidic fibroblast growth factor(aFGF) on human liver cell L-02 and human kidney cell 293 damaged by nitidine chloride in vitro. METHODS: The MTT assay was used to assess the proliferation of human liver cell L-02 and human kidney cell 293 treated with nitidine chloride.The contents of SOD and MDA and LDH in cultural supernate were measured by ultraviolet spectrophotometry. RESULTS: Nitidine chloride inhibited the proliferation of human liver cell L-02 and human kidney cell 293 in a dose-dependent manner. CONCLUSION: Nitidine chloride has certain toxicity on human liver cell L-02 and human kidney cell 293.aFGF could protect human liver cell L-02 and human kidney cell 293 damaged by nitidine chloride.
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AIM: To establish the quality standard of Rhinitis Compound Spray (Caulis Luffae, Herba Ephedrae, etc. ) . METHODS: TLC was used in qualitative identification of Caulis Luffae and Herba Ephedrae. And TLC-scanning was used for assay of oleanolic acid in the preparation. RESULTS: Caulis Luffae, Herba Ephedrae could be detected by TLC. Oleanolic acid had a good linear relationship within the range of 2.0-10.0 ?g (r= 0.999 8 ). The average recovery was 98.91%, and the RSD was 1.71%. CONCLUSION: The method established is stable and reliable. It can be used for quality control of the preparation.
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Alkaloid is one of the main antineoplastic activities in Chinese Herbal Medicine.There has been big progress on the mechanism of antitumor in natural alkaloids and their derivatives in recent years,especially focusing on the targets of hypoxia inducible factor 1,telomere,and topoisomerase.The new trends of these three targets have been reviewed in this paper.
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Aim To explore effects of acidic fibroblast growth factor(aFGF)on the growth of renal tubular epithelial cells cultured in vitro. Methods Renal tubular epithelial cells in rats were gained through digestion by pamcreatin. aFGF was added into cultures of renal tubular epithelial cells, as experimental groups. The number of cells was counted and the shape of cells was observed at 12,24,48 and 72 h. Results Renal tubular epithelial cells were successfully obtained from kidneys. After treated for 72 h,the renal tubular epithelial cells showed different proliferation in experimental and control groups. There was no obvious difference at 12 h, but there was statistical difference between the two groups at 24 h,48h and 72 h(P