1.Effect of Apigenin on Inhibiting Proliferation and Apoptosis of Human NCI-H1650 Cell Line
Xiaotong PAN ; Zhouping YANG ; Huan LIU ; Xuediao PAN
Herald of Medicine 2016;35(8):835-839
Objective To determine the effect of apigenin on inducing apoptosis and inhibiting proliferation of non-small cell lung cancer cell line NCI-H1650. Methods NCI-H1650 cell line was cultured routinely in vitro, with blank control group and different concentrations of apigenin (10, 20, 40, 80 μmol?L-1). The blank control group was RPMI-1640 solution without apigenin. Cell proliferation was detected by MTT. Hoechst 33258 was applied to observe morphological changes of the apoptotic cells after treatment of different concentrations of apigenin. Flow cytometry AnnexinV-FITC/PI double staining method was used to determine cell apoptosis rate. The expression levels of apoptosis-related signaling molecules Bax and Bcl-2 protein were performed by Western blotting. Results MTT showed that apigenin inhibited proliferation of NCI-H1650 cell line in a concentration-and time-dependent manner (P<0.01).Hoechst 33258 nuclear staining showed typical characteristics of apoptosis in certain concentration-dependent manner, such as nuclear condensation and appearance of apoptotic bodies. In addition, the results of flow cytometry staining indicated that the apoptotic rate of NCI-H1650 cells cultured with blank control group and different concentrations of apigenin (10, 20, 40, 80 μmol?L-1) for 48 h was (5.00±0.33)%, (18.05±4.67)%, (21.48± 1.95)%, (43.24±1.11)%, (66.23±3.65)%, respectively (P<0.01).Western blotting results showed that the expression levels of Bax increased with increasing of the drug concentration, but Bcl-2 decreased with increasing of the drug concentration. Conclusion Apigenin can inhibit the proliferation and induce apoptosis of NCI-H1650 cells in non-small cell lung cancer. Mechanisms may be related to increase of the expression of apoptosis related protein Bax and decrease of the expression of anti apoptosis protein Bcl-2.
2.Effects of Breviscapine on intracellular Ca~(2+) concentration in human umbilical vein endothelial cells
Xuediao PAN ; Banghao ZHU ; Li MA ; Jie LIU
Chinese Pharmacological Bulletin 1986;0(05):-
Aim To investigate the effects of Breviscapine(BE)on intracellular Ca2+ concentration in human umbilical vein endothelial cells(HUVECs). Methods HUVECs were incubated with the calcium ion-sensitive fluorescent indicator fluo-3/AM, and then a laser confocal microscope was applied to measure changes of fluorescence intensity under different agonists to investigate the effects of Breviscapine on intracellular Ca2+ concentration in HUVECs. Results ①BE induced Ca2+ transients in the buffer with or without Ca2+;②The Ca2+ transients induced by BE could be overlapped by cyclopiazonic acid (CPA);③Pretreatment of BE inhibited Ca2+ influx caused by KCl.④ BE had no evident effect on the calcium influx in- duced by the restoration of extracellular Ca2+ after the depletion of intracellular Ca2+ stores.Conclusion BE depletes CPA-sensitive intracellular Ca2+ stores and inhibits Ca2+ influx through voltage-dependent Ca2+ channels, and BE has no evident effect on the calcium influx induced by the restoration of extracellular Ca2+ after the depletion of intracellular Ca2+ stores.
3.Screening of tumor mark for the lung cancer by using peptide ZS-6
Wenbin FENG ; Xuediao PAN ; Jie ZHOU ; Lei SHI ; Junqiang LUO ; Xun LI ; Linquan ZANG
Chinese Pharmacological Bulletin 2010;26(1):44-47
Aim To select a potential biomarker for early lung cancer diagnosis and targeted therapy by using the cancer specific bounded peptide ZS-6 which had already been obtained from the laboratory.Methods The peptide ZS-6 marked by biotin was used as a probe to pan out the human lung cancer cDNA phage display library,after 4 rounds of subtraction panning,the specific binding clones of ZS-6 were identified.After amplification and purification,then those DNA sequences were identified and analyzed with bioinformatics.Results 18 phage clones were identified to the specific peptide ZS-6 and the DNA sequence showed one of them was an unknown new gene while the others were known tumor related genes.Conclusion A tumor biomarker selected from human lung cancer cDNA library provides a potential tool for early lung cancer diagnosis and therapy.
4.Effects of ERK1/2/PPARα/SCAD signal pathways on physiological car-diac hypertrophy and pathological cardiac hypertrophy
Qiuju HUANG ; Jinxian HUANG ; Jiani LUO ; Peiqing LIU ; Shaorui CHEN ; Xuediao PAN ; Linquan ZANG ; Sigui ZHOU
Chinese Journal of Pathophysiology 2014;(8):1427-1432
[ABSTRACT]AIM:ToinvestigatethedifferenteffectsofERK1/2/PPARα/SCAD(short-chainacyl-CoAdehy-drogenase) signal pathways on the cardiac hypertrophy induced by insulin-like growth factors 1 ( IGF-1) or phenylephrine ( PE) .METHODS:The neonatal rat cardiomyocytes induced by IGF-1 were used as the model of physiological cardiac hypertrophy , and those induced by PE were used as the model of pathological cardiac hypertrophy .The surface area of the cardiomyocytes, the expression of p-ERK1/2, PPARαand SCAD, the activity of SCAD and the content of free fatty acid in the cardiomyocytes were measured .RESULTS:Compared with the control cells , the surface area of the cardiomyocytes in-duced by IGF-1 and PE were both increased .Compared with the controls , the expression of SCAD and PPARα, and the activity of SCAD in the cardiomyocytes induced by IGF-1 were increased , while the expression of p-ERK1/2 was de-creased.However, the cardiomyocytes treated with PE showed decreased expression of SCAD and PPARα, decreased activ-ity of SCAD and increased expression of p-ERK1/2.Meanwhile, the decrease in free fatty acid in IGF-1-induced cardio-myocytes and the increase in PE-induced cardiomyocytes indicated that the fatty acid utilization was increased in the cardio -myocytes induced by IGF-1, but decreased in the cardiomyocytes induced by PE .CONCLUSION: The changes of p-ERK1/2, PPARαand SCAD in the cardiac hypertrophy induced by IGF-1 or PE indicate that the effects of ERK 1/2/PPARα/SCAD signal pathways are different between physiological cardiac hypertrophy and pathological cardiac hypertro -phy , and that SCAD may be a molecular marker of these 2 different cardiac hypertrophies and a potential therapeutic target for pathological cardiac hypertrophy .
5.Preparation and Characterization of Scutellarin Nanosuspension
Rentong LIU ; Xuediao PAN ; Xiaohui LUO ; Weisi LIN ; Wenting LIU ; Zhicheng YANG
China Pharmacist 2017;20(1):42-45
Objective: To prepare scutellarin nanosuspension ( SCU-NS) and study the main influencing factors in the prepara-tion. Methods:The technology parameters were determined, and then the influencing factors of SCU-NS were studied. The optimal formula was confirmed by orthogonal design with zeta potential as the evaluation index. Results: The optimal formula process was as follows:drug amount was 0. 5 g, Pluronic? F68 amount was 0. 1 g, phospholipid amount was 0. 2 g, SDS amount was 0. 05 g and HPMC E5 amount was 0. 05 g. The average particle size and the zeta potential of SCU-NS was (122 ± 4) nm and ( -25. 5 ± 0. 6) mV, respectively. The result of transmission electron microscope showed that SCU-NS was spherical and uniform, and the dissolution of SCU-NS in 30 min was more than 90%. Conclusion:Nanosuspension can significantly enhance the dissolution of SCU.
6.Effective components of Semen Ziziphi Spinosae for sedative-hypnotic based on receptor ligand binding assay
Jiahong LI ; Qiling TANG ; Jiquan GUO ; Qiyou WANG ; Shuguang ZHU ; Youdi WANG ; Chen YANG ; Siyuan GAO ; Jinyong LIU ; Ruiping GAO ; Junye WANG ; Xiaoqiong YE ; Xuediao PAN ; Linquan ZANG
Chinese Pharmacological Bulletin 2016;32(4):508-513
Aim To determine the effective compo-nents of Semen Ziziphi Spinosae for sedative-hypnotic and its mechanism. Methods The extraction of Se-men Ziziphi Spinosae and the rat brain homogenates were prepared. High concentrations of Diazepam com-petitively replaced the ligand compounds of Semen Ziz-iphi Spinosae combining BDZ receptor in brain tissue, and all the compounds with sedative and hypnotic effects were collected and identified by HPLC and LC-MS technique, as the compounds extracted from the brain tissue were administered with Semen Ziziphi Spi-nosae. The brain tissue was administered with Diaze-pam, and with Semen Ziziphi Spinosae and Diazepam. Results The HPLC chromatograms show that the peak time of BDZ receptor ligand compounds was 2. 71 min and 46. 87min, when compared with Diazepam. And the LC-MS chromatograms display the relative molecu-lar weight of the ligand compounds was 274. 28 m/z, 453. 34 m/z,496. 34 m/z and 608. 38 m/z respective-ly. According to the fingerprint of Semen Ziziphi Spi-nosae, these compounds may be fatty acid substances and lupine pill triterpene compounds. Conclusions On the basis of the principle of receptor ligand bind-ing, we established a way to quickly analyze and iden-tify the role of natural products in the same drug target compounds. The method not only can clearly define the effective components of natural products, but also clar-ify the mechanism of action of the compounds. The ac-tive ingredient of calm hypnosis in Semen Ziziphi Spi-nosae may be fatty acid substances Palmitic acid ( C16 H32 O2 ) and lupine pill triterpene compounds Alphitolic acid( C30 H48 O4 ) and Spinosin( C28 H32 O15 ) . They exert their sedative and hypnotic effects by combining with BDZ receptor, and the research has laid a theoretical foundation for the further study about mechanism of Se-men Ziziphi Spinosae.
7.Effect of apigenin on proliferation and apoptosis of human lung cancer NCI-H460 cells.
Xuediao PAN ; Zhouping YANG ; Zhiying YANG ; Sigui ZHOU ; Haitao ZHANG ; Linquan ZANG
Journal of Southern Medical University 2013;33(8):1137-1140
OBJECTIVETo study the effect of apigenin on the proliferation and apoptosis of human lung cancer cell line NCI-H460.
METHODSNCI-H460 cells were cultured with different concentrations of apigenin, and MTT assay was used to evaluate the cell inhibition rates. Apoptosis of NCI-H460 cells was observed under a fluorescence microscope with Hoechst 33258 staining and quantified by flow cytometry using annexin V-FITC/PI stain. The expressions of apoptosis-related proteins Bax, Bcl-2 and caspase-3 were analyzed by Western blotting.
RESULTSApigenin causes concentration- and time-dependent inhibition of the proliferation of the cells. NCI-H460 cells treated with apigenin showed significant morphological changes of apoptosis, and the cell apoptotic rates increased as apigenin concentration increased. Western blotting demonstrated that apigenin increased the protein levels of Bax and caspase-3 and reduced the protein expression of Bcl-2.
CONCLUSIONApigenin can inhibit the proliferation and induce apoptosis of NCI-H460 cells possibly by up-regulating expression of Bax and caspase-3 and down-regulating the expression of Bcl-2.
Apigenin ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Signal Transduction ; bcl-2-Associated X Protein ; metabolism
8.Change of short-chain acyl-CoA dehydrogenase in heart failure after myocardial infarction in rats and the intervention of aerobic exercise
Yingqin LIAO ; Zhonghong LI ; Zhaohui SHU ; Xiaoyi ZHONG ; Yongshao SU ; Zhichao MA ; Peiqing LIU ; Jing LU ; Linquan ZANG ; Xuediao PAN ; Sigui ZHOU
Chinese Critical Care Medicine 2019;31(2):172-177
Objective? To?Study?the?changes?of?short-chain?acyl-CoA?dehydrogenase?(SCAD)?in?heart?failure?(HF)?after?myocardial?infarction?(MI),?and?the?effect?of?aerobic?exercise?on?SCAD.? Methods? Healthy?male?Sprague-Dawley?(SD)?rats?were?divided?into?sham?operation?group?(Sham?group),?sham?operation?swimming?group?(Sham+swim?group),?HF?model?group?(LAD?group)?and?HF?swimming?group?(LAD+swim?group)?by?random?number?table?method,?with?9?rats?in?each?group.?The?left?anterior?descending?branch?of?coronary?artery?(LAD)?was?ligated?to?establish?a?rat?model?of?HF?after?MI.?In?Sham?group,?only?one?loose?knot?was?threaded?under?the?left?coronary?artery,?and?the?rest?operations?were?the?same?as?those?in?LAD?group.?Rats?in?Sham+swim?group?and?LAD+swim?group?were?given?swimming?test?for?1?week?after?operation?(from?15?minutes?on?the?1st?day?to?60?minutes?on?the?5th?day).?Then?they?were?given?swimming?endurance?training?(from?the?2nd?week?onwards,?60?minutes?daily,?6?times?weekly,?10?weeks?in?a?row).?Tail?artery?systolic?pressure??(SBP)?was?measured?before?swimming?endurance?training?and?every?2?weeks?until?the?end?of?the?10th?week.?Ten?weeks?after?swimming?training,?echocardiography?was?performed?to?measure?cardiac?output?(CO),?stroke?volume?(SV),?left?ventricular?ejection?fraction?(LVEF),?shortening?fraction?(FS),?left?ventricular?end-systolic?diameter?(LVESD),?left?ventricular?end-diastolic?diameter?(LVEDD),?left?ventricular?end-systolic?volume?(LVESV),?and?left?ventricular?end-diastolic??volume?(LVEDV).?Morphological?changes?of?heart?were?observed?by?Masson?staining.?Apoptosis?of?myocardial?cells?was?detected?by?transferase-mediated?deoxyuridine?triphosphate-biotin?nick?end?labeling?stain?(TUNEL)?and?apoptosis?index?(AI)?was?calculated.?Reverse?transcription-polymerase?chain?reaction?(RT-PCR)?and?Western?Blot?were?used?to?detect?the?mRNA?and?protein?expression?of?myocardial?SCAD?respectively.?In?addition,?the?enzyme?activity?of?SCAD,?the?content?of?adenosine?triphosphate?(ATP)?and?free?fatty?acid?(FFA)?in?serum?and?myocardium?were?detected?according?to?the?kit?instruction?steps.? Results? Compared?with?Sham?group,?Sham+swim?group?showed?SBP?did?not?change?significantly,?with?obvious?eccentric?hypertrophy?and?increased?myocardial?contractility,?and?LAD?group?showed?persistent?hypotension,?obvious?MI,?thinning?of?left?ventricle,?and?decreased?myocardial?systolic/diastolic?function.?Compared?with?LAD?group,?SBP,?systolic/diastolic?function?and?MI?in?LAD+swim?group?were?significantly?improved?[SBP?(mmHg,?1?mmHg?=?0.133?kPa):?119.5±4.4?vs.?113.2±4.5?at?4?weeks,?120.3±4.0?vs.?106.5±3.7?at??6?weeks,?117.4±1.3?vs.?111.0±2.3?at?8?weeks,?126.1±1.6?vs.?119.4±1.9?at?10?weeks;?CO?(mL/min):?59.10±6.31?vs.?33.19±4.76,?SV?(μL):?139.42±17.32?vs.?84.02±14.26,?LVEF:?0.523±0.039?vs.?0.309±0.011,?FS:?(28.17±2.57)%?vs.?(15.93±3.64)%,?LVEDD?(mm):?8.80±0.19?vs.?9.35±0.30,?LVESD?(mm):?5.90±0.77?vs.?7.97±0.60,?LVEDV?(μL):?426.57±20.84?vs.?476.24±25.18,?LVESV?(μL):?209.50±25.18?vs.?318.60±16.10;?AI:?(20.4±1.4)%?vs.?(31.2±4.6)%;?all?P?0.05].?Compared?with?Sham?group,?the?mRNA?and?protein?expression?of?myocardium?SCAD,?the?activity?of?SCAD?in?Sham+swim?group?were?significantly?increased,?the?content?of?ATP?was?slightly?increased,?the?content?of?serum?FFA?was?significantly?decreased,?and?the?content?of?myocardial?FFA?was?slightly?decreased;?conversely,?the?mRNA?and?protein?expression?of?myocardium?SCAD,?the?activity?of?SCAD?and?the?content?of?ATP?in?LAD?group?were?significantly?decreased,?the?content?of?serum?and?myocardial?FFA?were?significantly?increased.?Compared?with?LAD?group,?the?mRNA?and?protein?expression?of?myocardium?SCAD,?the?content?of?ATP?were?significantly?increased?in?LAD+swim?group?[SCAD?mRNA?(2-ΔΔCt):?0.52±0.16?vs.?0.15±0.01,?SCAD/GAPDH?(fold?increase?from?Sham?group):?0.94±0.08?vs.?0.60±0.11,?ATP?content?(μmol/g):?52.8±10.1?vs.?14.7±6.1,?all?P?0.05],?the?content?of?serum?and?myocardial?FFA?were?significantly?decreased?[serum?FFA?(nmol/L):?0.11±0.03?vs.?0.29±0.04,?myocardial?FFA?(nmol/g):?32.7±8.2?vs.?59.7±10.7,?both?P?0.05],?and?the?activity?of?SCAD?was?slightly?increased?(kU/g:?12.3±4.3?vs.?8.9±5.8,?P?>?0.05).? Conclusion? The?expression?of?SCAD?in?HF?was?significantly?down-regulated,?and?the?expression?was?significantly?up-regulated?after?aerobic?exercise?intervention,?indicating?that?swimming?may?improve?the?severity?of?HF?by?up-regulating?the?expression?of?SCAD.