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Objective To observe the effect of β-asarone on the proliferation, cycle, apoptosis and migration of tumor cells A549, PC3, and PC9-R, thus to provide experimental basis for the application of β-asarone to the treatment of cancer. Methods After A549, PC3, and PC9-R were cultured with different concentrations ofβ-asarone for 24 h, 48 h, and 72 h respectively, CCK-8 was used to detect the optical density (D) of cell proliferation, and then the cell proliferation rate was calculated. The flow cytometry was used to measure the cell DNA cycle and cell apoptosis, and Transwell Chambers were used to detect the cell migration. Results After treatment with different concentrations of β-asarone for 24 h, 48 h, and 72 h respectively, the growth of A549, PC3, and PC9-R showed declining trend in concentration- and time-dependent manner. The proportion of A549, PC3, and PC9-R at G0/G1 phase was increased, the proportion of the three kinds of cells at S phase and the proliferation indexes were declined, the apoptotic rate of A549, PC3, and PC9-R was increased, and the migration of A549, PC3, and PC9-R was reduced (P<0.05 or P<0.01 compared with those of the normal control group). Conclusion β-asarone has certain effects on suppressing proliferation, blocking G0/G1 phase developing into S phrase, inhibiting DNA synthesis, promoting the apoptosis, and inhibiting the migration of A549, PC3 and PC9-R.
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Objective To investigate the protective effect of Buyang Huanwu Decoction ( BYHWD) on PC12 cell apoptosis induced by oxygen-glucose deprivation ( OGD) and to screen out the main ingredients of BYHWD on protecting PC12 cell from injury induced by OGD. Methods Seven herbs of BYHWD were selected as the influencing factors, and they were Radix Astragali, Radix Paeoniae Rubra, the carda part of Radix Angelicae Sinensis, Pheretima Asiatica, Rhizoma Chuanxiong, Semen Persicae, and Flos carthami. Two levels were designed for each factor, and the experiment was arranged according to L8 ( 27) orthogonal test table. Apoptosis of PC12 cells induced by OGD was detected by serum pharmacology method and flow cytometry techniques. With apoptotic rate as the evaluation index, visual analysis and variance analysis were used for the comparison of apoptosis of PC12 cells in the blank control group ( oxygen + sugary medium + blank serum) , model group (oxygen-glucose-free medium + blank serum) and drug groups (oxygen-glucose-free medium +serum containing BYHWD or the separate recipe of BYHWD ) . Results Compared with the blank control group, apoptotic rate was increased in PC12 cells of the model group, the difference was statistically significant ( P<0.01) . Compared with the model group, the apoptotic rate was decreased in BYHWD group, and the differences was statistically significant (P<0.01) . The results of orthogonal analysis showed that Semen Persicae, Flos carthami and Radix Paeoniae Rubra were the main active ingredients of BYHWD on depressing apoptosis of PC12 cell induced by OGD ( P<0.01). The apoptotic rate of serum containing the prescription with Radix Paeoniae Rubra was lower than that of the prescription without Radix Paeoniae Rubra. Conclusion BYHWD could inhibit OGD-induced PC12 cell apoptosis, and Radix Paeoniae Rubra is the main active herb on depressing apoptosis in BYHWD.
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AIM:To investigate the inhibitory effect of ethanol extract of propolis (EEP) on oxidized low-den-sity lipoprotein ( ox-LDL )-induced macrophage apoptosis and the underlying molecular mechanisms . METHODS:RAW264.7 macrophages were pretreated with EEP (7.5, 15 and 30 mg/L), 4-phenylbutyric acid (PBA, 5 mmol/L) or diphenyleneiodonium ( DPI, 5μmol/L) for 1 h and then treated with ox-LDL (100 mg/L) or tunicamycin ( TM, 4 mg/L) for 24 h.The cell viability and apoptosis were determined by MTT assay and Annexin V-FITC apoptosis detection kit , re-spectively.The activity of superoxide dismutase (SOD), and the levels of reactive oxygen species (ROS) and malondial-dehyde (MDA) in the cells were measured.The protein levels of caspase-12, a proapoptotic molecule under endoplasmic reticulum stress ( ERS) , were examined by Western blot analysis .RESULTS:Like PBA ( an ERS inhibitor ) , EEP pro-tected RAW264.7 macrophages from ox-LDL-induced injury in a dose-dependent manner , as assessed by the increased cell viability and the decreased apoptotic rate .The decrease in cell viability and increase in apoptotic rate induced by TM , an ERS inducer, were also attenuated by EEP .Moreover, EEP suppressed ox-LDL-induced oxidative stress as revealed by the decreased generation of ROS and MDA as well as elevated SOD activity , which were similar to DPI , an oxidative stress in-hibitor.Furthermore, EEP significantly suppressed ox-LDL-or TM-induced activation of caspase-12.Similar results were observed in the cells pretreated with PBA or DPI and then treated with ox-LDL.CONCLUSION: EEP may protect RAW264.7 macrophages from ox-LDL-induced apoptosis and the mechanism is at least partially involved in the ability of EEP to suppress oxidative stress and subsequent activation of caspase -12.
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[ABSTRACT]AIM:ToinvestigatetheeffectofD4F,anapolipoproteinA-Imimeticpeptide,onoxidizedlow-density lipoprotein ( ox-LDL)-induced macrophage apoptosis and activation of caspase-12, a key molecule in endoplasmic reticulum stress ( ERS )-associated apoptotic pathway, and to elucidate the underlying molecular mechanisms. METHODS:RAW264.7 macrophages were pretreated with D4F (12.5, 25 and 50 mg/L), 4-phenylbutyric acid (PBA, 5 mmol/L) or diphenyleneiodonium ( DPI, 5 μmol/L) for 1 h and then treated with ox-LDL (100 mg/L) or tunicamycin ( TM, 4 mg/L) for 24 h.The cell viability and apoptosis were determined by MTT assay and TUNEL detection, respective-ly.The levels of malondialdehyde ( MDA) and reactive oxygen species ( ROS) in the cells and the activities of superoxide dismutase ( SOD) and nicotinamide adenine dinucleotide phosphate ( NADPH) oxidase were determined.The protein level of caspase-12 was examined by Western blot analysis.RESULTS: Similar to the ERS inhibitor PBA, D4F protected RAW264.7 macrophages from ox-LDL or TM ( an ERS inducer)-induced decrease in the viability and increase in apoptotic rate in a dose-dependent manner.Like DPI (an oxidative stress inhibitor), D4F significantly inhibited ox-LDL-induced ox-idative stress, as expressed by the decreased generation of ROS and MDA ( P<0.01) , the increased activity of SOD and the decreased activity of NADPH oxidase (P<0.05).Moreover, similar to PBA and DPI, D4F significantly suppressed ox-LDL-induced activation of caspase-12 in a concentration-dependent manner ( P<0.05) .Furthermore, D4F also inhibi-ted the caspase-12 activation induced by TM (P<0.05).CONCLUSION: D4F inhibits macrophage apoptosis induced by ox-LDL, and the mechanism is at least partially by reducing oxidative stress and inhibiting the activation of caspase-12.
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AIM:To explore the effect of β-asarone on vascular endotheliam and adhesion molecule expression of endothelium induced by β-amyloid peptide from Alzheimer's disease and to estimate the injury repair.METHODS:Cultured ECV304 cells were incubated with freshly solublizeal Aβ1-42 and the mixture of Aβ1-42 and β-asarone,the expression of three central adhesion molecules,CD106,CD62P,CE62E and Ca2+ concentration were examined and apoptosis was recorded by Flow eytometry.Test viability of cells by MTT methods.RESULTS:The results showed that in model group and treated group,ligation of endothelial CD106,CD62P,CE62E,markers for endothelial cell activation and Ca2+ concentration,leads to a lot of release.The livability decreased and the apoptosis increased.Further more,simultaneous treatment of ECV304 cells with β-asarone resulted in the decrease significandy in these three adhesion molecules described above and Ca2+ concentration as well as the livability upper and apoptosis lower.CONCLUSION:CD106,CD62P,CE62E,important inflammational factor of Aβ-induced endothelial injury,may be promotion of the inflammatory scade in vascular endothelial.β-asarone may protect ECV304 cell apoptosis by regulate Ca2+ and expression of cell surface markers.
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Objective To study the protective effects of ?-asarone on PC12 cells damage induced by Glutamate. Method The effects of ?-asarone on PC12 cells after Glutamate intoxication on morphology, extent of damage, livability, intracellular calcium concentration and apoptosis ratio were observed. Result Morphological changes, LDH leakage and intracellular calcium concentration increasing, and cell survival decreasing were observed in PC12 cells exposured to Glutamate. 7.5, 15, 30 ?g/mL?-asarone can increase cell survival, decrease LDH leakage. 15, 30 ?g/mL ?-asarone can reduce intracellular calcium concentration and apoptosis ratio. Conclusion ?-asarone prevents the toxicity of Glutamate, and it maybe attribute to its effect of anticalcium.
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Objective To study the changes of diastolic function of left ventricle by tissue Doppler imaging (TDI) and the serum level of endocrine factors,and analyse the relations between these factors and mass or diastolic function of left ventricle in patients with essential hypertension.Methods Sixty one patients with essential hypertension were divided into left ventricular hypertrophy(LVH) group and no left ventricular hypertrophy(NLVH) group.Twenty healthy subjects were considered as control group.The early(e) and late(a) diastolic maximal myocardial velocity and e/a of lateral wall motion of mitral valve annulus were recorded by TDI.The levels of atrial natritic peptide(ANP),brain natriuretic peptide(BNP),endothelium(ET),calcitonic gene related peptide(CGRP) and insulin like growth factor 1(IGF 1) were measured by radio immunology analysis.Results The e wave maximal velocity in LVH group was lower than that of NLVH group and control group [( 14.56 ? 7.83 ) cm/s,( 16.40 ? 0.66 ) cm/s,( 18.68 ? 3.78 ) cm/s,respectively],and a wave maximal velocity in LVH group was higher than that of NLVH group and control group [( 18.28 ? 9.60 ) cm/s,( 16.03 ? 5.88 ) cm/s ,( 14.53 ? 1.28 ) cm/s,respectively]; The e and a maximal velocity velocities in both LVH and NLVH groups had statistic differences with those of control group(P
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Objective To observe the protective effect of?-asarone,the active components from Rhizoma Acori Tarinowii,on human umbilical venous endothelial cell strain ECV304 injury induced by oxidized low-density lipoprotein(ox-LDL)and to investigate its influence on proliferation of vascular smooth muscle cells(VSMC). Methods The well-grown ECV304 cells were randomized into 5 groups:normal group,control group,high-, middle-and low-dose?-asarone groups.Except the normal group,ECV in the other groups were co-cultured with ox-LDL 25?g/mL,and the?-asarone groups were additionally with?-asarone 30,15 and 7.5?g/mL added respectively.Flow cytometry was used to detect the cell apoptotic rate and the mitochondrial membrane potential (MMP)of ECV304 as well as the proliferation index and the DNA content in different cell phases of VSMC. Results In the model group,the apoptotic rate of ECV was increased,MMP was decreased,and VSMC proliferation was promoted(P
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OBJECTIVE:To study the quality standards for Shenchang injection.METHODS:TLC was applied to identify the properties of Borneol and volatile oil of Rhizoma Acori Tatarinowii in the formula.HPLC was adopted to determine the content of ?-asarone in Shenchang injection.RESULTS:?-asarone showed good linear relationship in the range of 0.632 3～ 1.264 6? g.The average recovery of ammonium ?-asarone was 99.92%(RSD=0.91%).CONCLUSION:The standard can be used for the quality control of Shenchang injection.
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OBJECTIVE:To study the effect of different dosage of grassleaf sweetflag rhizome on central nervous system.METHODS:The effects of different dosage of grassleaf sweetflag rhizome on the hours of sleep and hypoxia live time of mice were observed,and the effects of which on water contents of brain tissues,apoptosis,endothelin(ET),malonaldehyde(MDA),erythrocuprein(SOD),glutathione peroxidase(GSH-Px),etc.in rats with cerebral ischemia were observed as well.RESULTS:Compared with the control group,different dosage of grassleaf sweetflag rhizome could shorten the sleeping time of mice and prolong the live time of hypoxia mice;high dosage of grassleaf sweetflag rhizome could remarkably decrease the water content of brain tissue and MDA level and inhibit the apoptosis of brain tissue cells while increase the activity of SOD and GSH-Px.CONCLUSION:High dosage of grassleaf sweetflag rhizome has the strongest protective effects to central nervous system,the effects of middle and low dosage of grassleaf sweetflag rhizome are lower than that of the high dosage.
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Gas chromatography combined with mass spectrography (GC - MS) was used to analyze the content of Muscone, a form of natural Moschus in high purity, with N2 as the carrier gas and isopsoralen as the internal standard. Results: The linear range of Muscone was from 0.056 to 0.451 g/L, regression equation: y=0. 3454 x-0. 1076,r=0.9999,the recovery was 97.99%and its RSD was 5.0%. RSD of within - day precision was 3.85% and RSD of day - to - day was 2.20% . The contents of Muscone from three batches of natural Moschus were 1.82% (RSD= 2.50%) ,3.33%(RSD = 1.36%) and 4.40%(RSD3.85%) respectively. The method is simple, rapid, reliable, and can be used as quality control for Muscone of Moschus and its preparation.
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Objective To observe the effect of ?-asarone combined with eugenol against PC12 cell injury induced by amyloid beta protein (A?25-35) and to explore the advantage of compatibility of monomers in herbs.Methods PC12 cells injury were induced by A?25-35.The morphological features and survival rate of PC12 cells as well as the changes of intracellular Ca2+concentration and NO concentration were observed after administration.Results The optimum combination was 10?mol/L ?-asarone+3 ?mol/L eugenol,which can decrease the concentration of intracellular Ca2+,inhibit the production of NO and increase the survival rate of PC12 cells.Conclusion The combination of effective components in Acorus tatarinowii has a better protective effect on A?25-35-induced PC12 cell injury than Acorus tatarinowii.
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[Objective] To analyze the main chemical constituents of decoction and concentrated decoction of Rhizoma Acori Tatarinowii (RAT) by gas chromatography-mass spectrometry (GC-MS). [Methods] RAT was decocted and concentrated in the pottery for two times and then 6 batches of the decoction and its concentrated decoction were analyzed by GC-MS. [Results] Five kinds of components in a higher amount were found in the first and second decoction of RAT, including: ?-asarone, ?-asarone, 2, 3-dihydro-3, 5-dihydroxy-6-methyl-4H-pyran-4-one, 5-hydroxymethylfurfural and acoramone. The contents of volatile components of ?-asarone and ?-asarone were lower while those of water-soluble components higher in the concentrated decoction of RAT. [Conclusion] The therapeutic effect of RAT is the co-action of the multiple components of RAT; the effect is related not only with the volatile components but also with the water-soluble components. Therefore, more attention should be paid to the difference of components in the concentrated decoction, which is generally used in the research of new Chinese herbal medicine, and in the clinically used decoction.
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Object To research the percutaneous absorption of ?-asarone in various solvents in vitro. Methods To measure the concentration of ?-asarone passed through rat skin in various solvents which contained borneol and azote-ketone by HPLC, count the penetration of accumulation within 24 h and steady velocity. Results 0.1% Borneol could not promote the penetration of ?-asarone; azote-ketone could decrease the penetration of ?-asarone. The penetration of accumulation within 24 h and steady velocity of ?-asarone in 20% ethanol were (352.624?87.049) ?g/cm2, (18.902?4.840) ?g/(cm2?h), respectively. Conclusion The percutaneous absorption of ?-asarone is the best when the solvents contain no promoter.
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Objective: To determine the component of naphtha from Acorus tatarinowii Schott.which can pass through the blood-brain barrier. Methods: Naphtha in rat brain was analyzed by GC-MS after gastric infusion of naphtha. Results: The methylisoeugenol,elemicin, ?-asarone and ?-asarone were detected in rat brain. Conclusion: The resuscitative effect of naphtha is resulted from the comprehensive action of multiple components.
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Objective To investigate the effects of ?-asarone,the major constituent of Acorus tarinowii schott,on the expression of endothelial cell adhesion molecules induced by hypoxia and reoxygenation.Methods The expression rates of endothelial cell surface adhesion molecules VCAM-1 and CD62E were detected by flow cytometry(FCM) and specific labeled monoclonal antibody.Results Compared to the normal group,the expression rates of VCAM-1 and CD62E were increased significantly in the model group(hypoxia-reoxygenation group)(P
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AIM:To explore the effect of ?-asarone on vascular endothelium and adhesion molecule expression of endothelium induced by ?-amyloid peptide from Alzheimer's disease and to estimate the injury repair.METHODS:Cultured ECV304 cells were incubated with freshly solublized A?_ 1-42 and the mixture of A?_ 1-42 and ?-asarone,the expression of three central adhesion molecules,CD106,CD62P,CE62E and Ca 2+ concentration were examined and apoptosis was recorded by Flow cytometry.Test viability of cells by MTT methods.RESULTS:The results showed that in model group and treated group,ligation of endothelial CD106,CD62P,CE62E,markers for endothelial cell activation and Ca 2+ concentration,leads to a lot of release.The livability decreased and the apoptosis increased.Further more,simultaneous treatment of ECV304 cells with ?-asarone resulted in the decrease significantly in these three adhesion molecules described above and Ca 2+ concentration as well as the livability upper and apoptosis lower.CONCLUSION:CD106,CD62P,CE62E,important inflammational factor of A?-induced endothelial injury,may be promotion of the inflammatory scade in vascular endothelial.?-asarone may protect ECV304 cell apoptosis by regulate Ca 2+ and expression of cell surface markers.
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Objective: To establish the quality control standards of two intermediate products of Xingshen Nasal Drops:musk aromatic water and volatile oil of Acorus tatarinowii Schott so that the content of main components and composition of the different batches of ultimate products remained stable.Methods: The content limit of muskone of musk aromatic water was ascertained by GC-MS. And the characteristic finger print of volatile oil of Acorus tatarinowii Schott was established. Results: The content of muskone of musk aromatic water should be controlled at the range of 0.166～0.202mg/mL. RSD of muskone content of 3 batches of ultimate products was 4.49%, not larger than 10%. The characteristic finger point of volatile oil of Acorus tatarinowii Schott was composed of 6 main component peaks: Methyleugenol, cis-Methylisoeugenol, trans-Methylisoeugenol, Elemicin, ?-Asarone and ?-Asarone. This characteristiz finger print could be repeated when the ultimate product was determined. Conclusion: The establishment of quality control standard of intermediate product can raise the product quality of compound preparations. And GC-MS is one of effective determination methods.
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AIM: To study the antiepileptic effect of Annao Tablet (?-asarone extracted from Acorus tatarinowii schott). METHODS: The medicine was administered orally. The effects were observed in mouse's convulsion models induced by picrotoxin, thiosemicarbazide, maximal electroshock seizure (MES) and Penicillin G. RESULTS: Annao Tablet had obvious effect against convulsion induced by picrotoxin and thiosemicarbazide, postponed the delitescence, reduced the convulsion times, prolonged the survival time; antagonized MES. The medicine could depress the epileptic degree, postpone spasm latent period, reduce the wet dog shack times. CONCLUSION: Annao Tablet has obvious antiepileptic effect.