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Currently, the domestic and foreign research methods for the qualitative basis of Chinese materia medica (CMM) mainly include phytochemical separation method, pharmacological activity tracer method, chromatography, etc. The above methods have played an active role in elucidating the qualitative basis of CMM. However, how to form a multi-component research model that can better reflect the "holistic view" characteristics of CMM and reflect the effectiveness of CMM has become an important direction for scholars in the industry to explore. Serum spectrum-effect of CMM is based on the theory of spectrum-effect to study the relationship between serum chemical fingerprints and pharmacodynamics in vivo. In this paper, by consulting relevant literature, the research idea of serum spectrum-effect was collected, the research results were summarized, and the research bottleneck was analyzed and discussed, so as to provide reference for revealing the material basis of CMM.
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Objective: To investigate the effect of Liuwei Dihuangwan on connexin 32 (Cx32) in hepatoma cell line CBRH7919 and its gap junction intercellular communication (GJIC), and furthermore study its mechanism of enhancing the bystander killing effect of suicide gene therapy. Method: Liuwei Dihuangwan (32 g·kg·d-1) and the same volume of normal saline were given to the rats by intragastrical administration. Blood was taken to prepare the medicated serum of Liuwei Dihuangwan and blank control serum, respectively. The hepatoma cell line CBRH7919 were treated by control serum and medicated serum of Liuwei Dihuangwan in different concentrations. There were four groups in experiment:the blank control group (volume fraction of 10%), medicated serum high dose group of Liuwei Dihuangwan (the volume fraction of 10%), medicated serum middle dose group of Liuwei Dihuangwan (the volume fraction of 5%), and medicated serum low dose group of Liuwei Dihuangwan (the volume fraction of 2.5%). The expression levels of Cx32 protein and mRNA in hepatoma cell line CBRH7919 were detected by indirect immunofluorescence assay (ⅡA) and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) assay. The fluorescence redistribution after photobleaching (FRAP) method was used to detect the function of GJIC of hepatoma cell line CBRH7919. Result: ① The indirect immunofluorescence assay (ⅡA) analysis indicated that as compared with the blank control group, the cx32 expression of CBRH7919 cells was up-regulated in a concentration-dependent manner in each dose group of the serum containing Liuwei Dihuangwan (PPPPConclusion: The mechanism of medicated serum of Liuwei Dihuangwan in enhancing the bystander killing effect of suicide geneis related to gap junction. Liuwei Dihuangwan may enhance the function of GJIC by increasing the localization of cx32 on the cell membrane of CBRH7919 cells and increasing the expression of cx32 mRNA and protein to achieve the synergistic action.
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Objective@#To investigate the effect of Yanghe decoction serum on the proliferation of breast cancer stem cells MDA-MB-231 and the expression of Smad3 and NF-κB.@*Methods@#According to the random number table method, 20 female SD rats were divided into blank control group and Yanghe decoction high, medium and low dose group. Drug serum were given gastric gavage of Yanghe decoction 28, 14, 7 g/kg, and control group were given gastric gavage of same volume saline. Each group of rats was given orally for 3 days to prepare 10% Yanghe decoction serum and blank serum. MDA-MB-231 cells were divided into blank control group and Yanghe decoction low, medium and high dose group according to random number table method. Low, medium and high dose Yanghe decoction groups were cultured with medium containing 10% Yanghe decoction high, medium and low dose drug serum, and the blank control group was cultured with medium containing 10% control serum. After drug intervention, the effects of each group on the proliferation of MDA-MB-231 cells at 24 h, 48 h, and 72 h were detected by MTT assay. The expression of Smad3 and NF-κB protein in each group was detected by Western blot. The expression of Smad3 and NF-κB mRNA in each group were detected by real-time fluorescence quantitative PCR.@*Results@#Compared to the control group, Yanghe decoction low, medium and high dose group of cells for 24 h (1.143 ± 0.093, 0.953 ± 0.069, 0.874 ± 0.041 vs. 1.239 ± 0.160), 48 h (1.223 ± 0.129, 0.989 ± 0.093, 0.864 ± 0.105 vs. 1.389 ± 0.095), 72 h (1.092 ± 0.147, 0.881 ± 0.095, 0.719 ± 0.086 vs. 1.353 ± 0.150) proliferation activity were significantly lower (P<0.05 or P<0.01). After 48 h of drug intervention, the expression of Smad3 protein (0.974 ± 0.098, 0.844 ± 0.084, 0.789 ± 0.105 vs. 1.214 ± 0.012), NF- κB p50 protein (0.994 ± 0.047, 0.911 ± 0.015, 0.765 ± 0.084 vs. 1.147 ± 0.103) in the low, medium and high dose Yanghe decoction group were significantly lower than those in the control group (P<0.05 or P<0.01). The expression of Smad3 mRNA (1.223 ± 0.129, 0.989 ± 0.093, 0.864 ± 0.105 vs.1.389 ± 0.095), NF-κB mRNA (1.092 ± 0.147, 0.881 ± 0.095, 0.719 ± 0.086 vs. 1.353 ± 0.150) significantly decreased in the Yanghe decoction group (P<0.05 or P<0.01).@*Conclusions@#Yanghe decoction can inhibit the proliferation of breast cancer stem cell MDA-MB-231, and lower the expression of Smad3 and NF-κB.
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The aim of this paper was to investigate the molecular mechanism of Calculus Bovis Sativus( CBS) in alleviating lipid accumulation in vitro by serum pharmacology. The CBS-containing serum of mice was obtained by serum pharmacology method to evaluate its effect on the proliferation of LO2 hepatocytes. The lipid reducing effects of CBS-containing serum through Nrf2 was evaluated by fructose-induced LO2 hepatocyte steatosis model,nuclear factor erythroid 2 related factor 2( Nrf2) agonist oltipraz combined intervention,cell oil red O staining and intracellular triglyceride( TG) content. The effects of CBS-containing serum on lipid peroxidation and hepatocytes apoptosis were evaluated by reactive oxygen species( ROS) and apoptosis assay,respectively. Real-time quantitative polymerase chain reaction( PCR) was used to detect the relative expression of lipid synthesis-related genes and apoptosis-related genes.RESULTS:: showed that CBS drug-containing serum had no significant effect on LO2 hepatocyte proliferation. As compared with the model group,CBS-containing serum could effectively reduce the formation of lipid droplets in fructose-induced LO2 hepatocytes,significantly reduce intracellular TG and ROS levels,and significantly reduce hepatocyte apoptosis rate( P < 0. 05). As compared with the model group,carbohydrate responsive element binding protein( ChREBP),sterol regulatory element binding protein-1 c( SREBP-1 c),fatty acid synthase( FAS),acetyl-CoA carboxylase 1( ACC1),stearoyl-CoA desaturase 1( SCD1),Bax and caspase-3 mRNA levels were significantly reduced in CBS drug-containing serum treatment group( P<0. 05). All of the above effects could be reversed by oltipraz.In conclusion,CBS-containing serum can significantly inhibit the fructose-induced LO2 liver fat deposition,and the mechanism may be related to reducing intracellular ROS level through the Nrf2 pathway and improving intracellular peroxidation state to reduce apoptosis.
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Animals , Cattle , Mice , Apoptosis , Cells, Cultured , Fatty Liver , Fructose , Gallstones , Chemistry , Hepatocytes , Cell Biology , Metabolism , Lipid Metabolism , Lipid Peroxidation , Liver , Medicine, Chinese Traditional , Reactive Oxygen Species , Metabolism , Serum , Chemistry , Sterol Regulatory Element Binding Protein 1 , Metabolism , TriglyceridesABSTRACT
Objective To investigate the effect of Yanghe decoction serum on the proliferation of breast cancer stem cells MDA-MB-231 and the expression of Smad3 and NF-κB. Methods According to the random number table method, 20 female SD rats were divided into blank control group and Yanghe decoction high, medium and low dose group. Drug serum were given gastric gavage of Yanghe decoction 28, 14, 7 g/kg, and control group were given gastric gavage of same volume saline. Each group of rats was given orally for 3 days to prepare 10% Yanghe decoction serum and blank serum. MDA-MB-231 cells were divided into blank control group and Yanghe decoction low, medium and high dose group according to random number table method. Low, medium and high dose Yanghe decoction groups were cultured with medium containing 10% Yanghe decoction high, medium and low dose drug serum, and the blank control group was cultured with medium containing 10% control serum. After drug intervention, the effects of each group on the proliferation of MDA-MB-231 cells at 24 h, 48 h, and 72 h were detected by MTT assay. The expression of Smad3 and NF-κB protein in each group was detected by Western blot. The expression of Smad3 and NF-κB mRNA in each group were detected by real-time fluorescence quantitative PCR. Results Compared to the control group, Yanghe decoction low, medium and high dose group of cells for 24 h (1.143 ± 0.093, 0.953 ± 0.069, 0.874 ± 0.041 vs. 1.239 ± 0.160), 48 h (1.223 ± 0.129, 0.989 ± 0.093, 0.864 ± 0.105 vs. 1.389 ± 0.095), 72 h (1.092 ± 0.147, 0.881 ± 0.095, 0.719 ± 0.086 vs. 1.353 ± 0.150) proliferation activity were significantly lower (P<0.05 or P<0.01). After 48 h of drug intervention, the expression of Smad3 protein (0.974 ± 0.098, 0.844 ± 0.084, 0.789 ± 0.105 vs. 1.214 ± 0.012), NF-κB p50 protein (0.994 ± 0.047, 0.911 ± 0.015, 0.765 ± 0.084 vs. 1.147 ± 0.103) in the low, medium and high dose Yanghe decoction group were significantly lower than those in the control group (P<0.05 or P<0.01). The expression of Smad3 mRNA (1.223 ± 0.129, 0.989 ± 0.093, 0.864 ± 0.105 vs.1.389 ± 0.095), NF-κB mRNA (1.092 ± 0.147, 0.881 ± 0.095, 0.719 ± 0.086 vs. 1.353 ± 0.150) significantly decreased in the Yanghe decoction group (P<0.05 or P<0.01). Conclusions Yanghe decoction can inhibit the proliferation of breast cancer stem cell MDA-MB-231, and lower the expression of Smad3 and NF-κB.
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Objective To observe the changes of iNOS, COX-2 and NF-κB in breast cancer MCF-7 cells treated with Yanghe decoction containing serum. Methods The MCF-7 human breast cancer cells were divided into blank group, low dose group, middle dose group and high dose group (4.5, 9, 18 g/kg, respectively). Real-time RT-PCR was used to detect the mRNA expression of iNOS and COX-2 at 24, 48, 72 and 96 hours, and Western-blot was used to detect the expression of NF-κB protein. Results Compared with the blank group, the expression of iNOS and COX-2 mRNA in the low dose group, middle dose group and high dose group decreased significantly at 24, 48, 72 and 96 hours(P<0.05), the the expression of NF-κB protein in the low dose group, middle dose group and high dose group decreased significantly at 24, 48, 72 and 96 hours (P<0.05). So it reduced with the increase of drug concentration and time. There was no significant difference in 72 hours after intervention. Conclusions The Yanghe decoction could reduce the expression of NF-κB, and then reducing the related inflammatory factors COX-2 and iNOS.
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Objective To study the influences of Yang-warming, Blood-activating, and Toxin-removing Recipe(YBTR)-containing serum sampled under different conditions on the proliferation of human umbilical vein endothelial cells(HUVECs). Methods HUVECs were cultured in vitro for the experiment. Pharmacological serum for the experiment was prepared as follows: Thirty-two SD rats (male in half) were randomly divided into 4 groups, namely normal saline group and low-, middle-, and high-dose YBTR groups (at the intragastric dosage of 10.35, 31.05, 93.15 g·kg-1 respectively, twice a day). The pharmacological serum was taken from one female rat and one male rat in various groups at 4 time points, i.e. at hour 1, 2 after first intragastric administration on the fourth feeding day, and at hour 1, 2 after first intragastric administration on the sixth feeding day(abbreviated as D3H1, D3H2, D5H1, D5H2 respectively). The effects of YBTR-containing serum on the proliferation of HUVECs were observed by CCK-8 assay method. Results The difference of proliferation-inhibition rate of HUVECs was statistically significant after treated with YBTR-containing serum prepared from rats of different genders at different time(Pgender=0.000<0.01, Ptime=0.000<0.01). The difference of interaction of time and gender was also significant (Ptime × gender=0.001<0.01), and the effect at D3H1 and D5H1 varied with the gender (PD3H2×gender = 0.000 < 0.01, PD5H1×gender = 0.002 < 0.01). The inhibitory action of YBTR-containing serum became stronger with the increase of the dosage of serum of female rat at D3H2, and the inter-group difference was statistically significant (P < 0.05), the effect showing concentration-dependent tendency. The inhibition of HUVECs proliferation reached the peak after treated by various doses of YBTR-containing serum from the female rat at D3H2, while the inhibition arrived to the peak after treated by low- and middle-dose YBTR-containing serum from the male rat at D5H1, and the inhibition arrived to the peak in the group of high-dose YBTR-containing serum from the male rat at D3H1. Conclusion The inhibitory action of YBTR-containing serum on the proliferation of HUVECs was stronger when the serum was taken from the female rat at D3H2.
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Objective To investigate the effect of Liuwei Dihuang Pills (LDP)on the expression of connexin 43 (Cx43) in rat hepatocarcinoma CBRH7919 cells. Methods LDP-containing serum was prepared by serum pharmacological method. The protein expression of Cx43 in CBRH7919 cells was determined by Western blotting method, indirect immuno-fluorescent staining labeled by fluorescein isothiocyanate (FITC)-laser scanning confocal microscope technology and flow cytometry. And the mRNA expression of Cx43 in the hepatoma cells was detected by reverse transcription real-time quantitative polymerase chain reaction(RT-qPCR). Results Compared with the blank serum control group,the protein and mRNA expression levels of Cx43 were increased by volume fraction of 2.5%, 5%, 10% LDP-containing serum (P < 0.05 or P < 0.01). Conclusion LDP play an anti-cancer role through promoting the expression of Cx43 in CBRH7919 cells, altering Cx43 membrane location, and improving the gap junctional intercellular communication(GJIC).
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Objective:To explore the effects of serum containing hemerocallis citrine baroni flavonids (HCBF)on the proliferation and apoptosis of human hepatoma HepG2 cells,and to clarify their mechanisms.Methods:The HepG2 cells were divided into blank control group, low (900 mg · kg-1 ), middle (1 800 mg · kg-1 )and high (2 700 mg·kg-1 )doses of serum containing HCBF groups.The inhibitory rates of growth of HepG2 cells were measured with MTT assay;inverted microscope was used to observe the morphologic changes;Annexin Ⅴ-PI double staining and flow cytometry were used to detect apoptotic rates of HepG2 cells; enzyme linked immunosorbent assay was used to detect the expression levels of Bax,bcl-2,and Caspase-3.Results:Compared with blank control group,the inhibitory rates of growth of HepG2 cells in middle and high doses of HCBF groups were increased (P <0.05 or P <0.01).The cells were shrank with irregular shape,and the nucleus was condensed and broken in HCBF group under inverted microscope.Compared with blank control group,the apoptotic rates of HepG2 cells in middle and high doses of HCBF groups were increased (P < 0.05 or P < 0.01).Compared with blank control group,the expression levels of bcl-2 protein in middle and high doses of HCBF groups were decreased (P <0.05 or P < 0.01),and the expression levels of Bax protein were significantly increased (P < 0.05 or P <0.01),and the expression level of Caspase-3 protein in high dose of HCBF group was significantly increased (P <0.05).Conclusion:The serum containing HCBF could induce the apoptosis of HepG2 cells,and its mechanism may be related to mitochondrial pathway.
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Objective To investigate the effects of Xinshang Xuduan Decoction(XXD, mainly composed of Rhizoma Drynariae, Radix Dipsaci, and Eupolyphaga seu Steleophaga) on the proliferation and differentiation of rat osteoblasts and bone morphogenetic protein 2(BMP-2)expression in the osteoblasts . Methods The animal serum containing XXD was prepared by serum pharmacological method and then was mixed together with α-MEM for the cell culture. Osteoblasts were isolated from the skull bone of SD neonatal rats by collagenase digestion and were identified by alkaline phosphatase(ALP) staining and alizarin red staining. The third and fourth generations of osteoblasts were treated with XXD at the volume fraction of 5%, 10%, 20% for 24, 48 , 72 hours respectively, and then the proliferation of the cells was evaluated by Cell Counting Kit-8(CCK-8). In the other test, osteoblasts were cultured with blank control serum, and 10% serum containing XXD, Radix Dipsaci, and Rhizoma Drynariae, respectively, and then the ALP activity was examined by using ALP assay kit and the expression of BMP-2 was investigated by enzyme-linked immunosorbent assay(ELISA). Results XXD had a dose- and time-dependent effect on the proliferation of rat osteoblasts in a suitable volume fraction range from 5% to 20%, and the effect of XXD at 10% was the best. Compared with the blank control serum group, ALP activity was increased in the cells treated with 10% serum containing XXD, Radix Dipsaci, and Rhizoma Drynariae(P<0.01). On culturing day 7, the expression of BMP-2 was increased in 10% XXD group and Rhizoma Drynariae group(P<0.05). Conclusion XXD can increase the ALP activity and BMP-2 expression in the osteoblasts in vitro, so does the single herb of Rhizoma Drynariae. And their therapeutic mechanism in promoting the healing of fractures may be related with the enhancement of osteogenesis of osteoblasts.
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Objective: To study the in vivo pharmacodynamics characteristics of Buyang Huanwu Decoction (BHD) in mice, and to provide reference for rational clinical drug use. Methods: The level of MDA and activity of SOD in liver tissue of mice were determined after ig treatment with BHD or its drug-containing serum. Further more, the time-effect and dose-effect relationship of BHD on MDA level and SOD activity were carried out. The parameters of pharmacodynamics were estimated based on the time-effect and dose-effect curves. Results: Compared with control group, both BHD and its drug-containing serum could reduce MDAlevel, while raise SOD activity in the liver of mice (P < 0.05). Time-effect curves both present multi-peak but most pharmacodynamics parameters showed significant differences (P < 0.05), except tp and t1/2(Ka). Conclusion: BHD has obvious antioxidant effect and pharmacodynamics characteristics can be described as one-compartment model.
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This review generalized and analyzed the studies on serum pharmacology and serum pharmacochemistry in Chinese materia medica (CMM) in recent years, mainly including the relationship between efficacy and serum, the preparation of serum containing drugs, and the significance of serum pharmacochemistry on improvement of CMM quality control standards. We summarized the advantages and disadvantages of serum pharmacology and serum pharmacochemistry in in vitro studies on the efficacy, and further envisaged the application of serum fingerprint combined with microdialysis sampling techniques to promote the development of serum pharmacology and serum pharmacochemistry.
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The preparation of Chinese materia medica (CMM) possesses the characteristic of holistic function involving multi-component, multi-target, and multi-link. This characters make it difficult to optimize the preparation process and construct the quality standards of CMM. Thus process building and quality evaluation system based on correctly identifying the components that closely related to the efficacy is a key scientific issue in the preparation process design and quality control of CMM. In this paper, a more comprehensive analysis on the feasibility of different research methods was carried out, including the spectral-efficiency relationships, pharmacokinetics, serum medicinal chemistry and serum pharmacology of drugs, the effect of integration effect correlation analysis, virtual screening, and comparative analysis of knock-in/knock-out. The purpose of this article is to provide the reference for identifying the bioactive marker group which is used for the CMM preparation process and quality evaluation.
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Objective Using serum pharmacology method to assess the optimal dose of Rhodiolasachalinensis extract (RSE)and Codonopsispilosula extract (CPE)combination as well as the effects of the combination on the immune function of mice.Methods Using the 3×3 facterial design,nine medicated serum of CPE and RSE mixture were made to assess the optimum combinational dose of CPE and RSE by detecting the T,B lymphocyte proliferation abilities and NK cell activity in vitro .Using the optimum combinational dose and reducing 2.5,5 and 10 times as high,middle and low doses of RSE+CPE groups,and the T,B lymphocyte proliferation abilities and the activity of NK cells in the mice were detected in vivo .Results The result of serum pharmacology indicates that compared with control group,the proliferation abilities of T,B lymphocytes and the activity of NK cells inducing by ConA and LPS in 200 mg· kg-1 RSE + 790 mg· kg-1 CPE group were increased (P < 0.05).The result of in vivo experiment indicated that compared with cyclophosphamide group,the spleen indexes (SI)in middle and high doses of RSE+CPE groups were significantly increased (P <0.05);compared with low dose of RSE+CPE group,the WBC number in middle dose of RSE + CPEgroup was significantly increased (P < 0.05 ). Compared with cyclophosphamide group,the T and B cell proliferation abilities induced with ConA and LPS and killing activities of NK cells in low,middle and high doses of RSE+CPE groups and positive drug CVT-200 group were significantly increased (P <0.05).Compared with CVT-200 group,low and high doses of RSE+CPE groups,the T and B cell proliferation abilities induced with ConA and LPS and the activity of NK cells in middle dose of RSE+CPE group were significantly increased (P <0.05).Conclusion RSE and CPE mixture can enhance the immune function of mice;RES 200 mg·kg-1 and CPE 790 mg·kg-1 are the optimal doses.
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Tong-Mai(TM) granules were composed ofRadix Salviae Miltiorrhizae(Danshen),Rhizoma Chuanxiong(Chuanxiong) andRadix Puerariae(Gegen). It had the effect of activating blood circulation. It had been used to treat ischemic cardio-cerebrovascular diseases in the clinical practice. This research combined serum pharmacochemistry and serum pharmaocology to study the material basis of active components in TM granules. After single or multiple intragastric administrations of TM granules, serum blood samples of rats were collected at different time points. LC-MS/MS method was developed to analyze chemical components of TM in blood serum samples. The cardiomyocyte hypoxia / reoxygenation (H/R) model was used in the evaluation of cardiomyocyte protection by TM. The correlation analysis was also conducted between serum concentration of TM and cardiomyocyte activity. The results showed that 8 components of pueraria flavonoid, 5 components of salvianolic acids and 2 components fromGegen were promptly absorbed and reached their highest concentrations at 5 or 30 min after administration. After 3 times of medication, the serum concentration was obviously higher compared to single medication. The drug-serum of TM showed significant protective effect on the cardiomyocyte H/R injury with dose-effect relationship. Daidzein, lithospermic acid, salvianolic acid A, salvianic acid A and rosmarinic acid presented as the most correlated components linked to the effect of activating blood circulation by TM. The serum pharmacochemistry / serum pharmacology related studies provided references for the verification of material basis of active components in compound Chinese medicine.
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BACKGROUND: Chinese medicine Gukang prescription has a clear effect on clinical treatment of osteoporosis, but the therapeutic pathway is stil unclear. OBJECTIVE:To investigate the effects of Chinese medicine Gukang on the expression of receptor activator of nuclear factor kappa B ligand and osteoprotegerin by regulating core binding factor alpha 1 expression to control the growth and development of osteoblasts. METHODS:Sprague-Dawley neonatal rats within 24 hours after delivery were used for the separation and culture of osteoblasts. Adult Sprague-Dawley rats were used to prepare drug-containing serum, and then divided into two groups randomly:normal control group and Gukang group. Rats in the normal control and Gukang groups were intragastrical y administrated with extract of Gukang prescription and normal saline based on rat’s body surface area, for 1 consecutive week. Two hours after the last administration, blood samples were taken from the heart. Then the serum was col ected. Osteoblasts at passage 3 were confirmed with alkaline phosphatase assay and digested. After counting and planting, al osteoblasts were divided into two groups and treated with col ected serum for 72 hours. Proliferative rate of osteoblasts was detected by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test. Secretion of alkaline phosphatase was detected by using enzyme linked immunosorbent assay and corrected with the corresponding absorbance value. mRNA expression of core binding factor alpha 1, receptor activator of nuclear factor kappa B ligand and osteoprotegerin were detected by using reverse transcription-PCR in al groups. RESULTS AND CONCLUSION:mRNA expression of osteoprotegerin and core binding factor alpha 1 in the Gukang group was significantly higher than that in the normal control group, but protein and mRNA expression of receptor activator of nuclear factor kappa B ligand were dramatical y lower in the Gukang group compared with the normal control group (Pcore binding factor alpha 1, thereby adjusting the expression of receptor activator of nuclear factor kappa B ligand and osteoprotegerin, which may be one of the mechanisms underlying Gukang treatment for osteoporosis.
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[Objective] To observe the inhibiting capacity of Bingcha Shuan Containing serum to HEPG2 cells' reproductive activity and to conjecture.Bingcha Shuan has the potential to cure the liver carcinoma.[Method]According to serum pharmacology,HEPG2 cells were cultured in vitro and interfered by various concentrations of Bingcha Shuan contained serum for 24,48 hours respectively.The inhibiting capacity of the Bingcha Shuan contained Serum to HEPG2 cells were detected by MTT.[Result] The Bingcha Shuan contained Serum has the inhibiting capacity to HEPG2 cells in some degree.And there was a marked positive correlation between drug concentration,time and inhibition ratio.[Conclusion] It is suggested that Bingcha Shuan has the inhibiting capacity of cell proliferation and it can provide the support on how to cure the liver cancer of clinic.
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@#Objective To investigate the inhibitory effect of Pingliukang (a prescription) medicated serum on the proliferation of cultured rat C6 glioma cells and influence on cell cycle in vitro. Methods MTT colorimetry were performed to measure the levels of the proliferation of rat C6 glioma cells cultured with 2.5%, 5%, 10% and 20% of Pingliukang medicated serum for 24 h, 48 h and 72 h in vitro. The effect of Pingliukang medicated serum on cell cycle were observed by FCM. Results The proliferation of C6 cell was obviously inhibited by Pingliukang medicated serum with dose-effect relationship. The inhibitory effect of 20% of medicated serum was the strongest. When the C6 glioma cells were treated with 10% and 20% medicated serum for 48 h and 72 h, the cells of S period declined. Conclusion The Pingliukang medicated serum can inhibit the proliferation and block cell cycle of cultured C6 glioma cells.
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Objective To study the influence of serum containing Ziqi Ruangan Pill on inhibitory effect of alcohol LO2 and to investigate the regulation on apoptosis related genes and the protective mechanism for LO2. Methods Rabbit sera saline containing Ziqi Ruangan Pill was prepared. The apoptotic LO2 model was established by being cultured with alcohol. The effect of serum on the model was detected by MTT method. Identification of apoptosis was carried out using flow cytometry and Hoechst3325 fluorescent staining. The expression of apoptotic inducing gene,casepase3 and NF-?B was detected by Real-time RT PCR and Western-Blot. Results The serum containing Ziqi Ruangan Pill could regulate the expression of the casepase3,NF-?B and reduce the apoptosis. Conclusion Ziqi Ruangan Pill can protect the LO2 by regulating the expression of apoptosis stimulative gene casepase3 and NF-?B.
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Objective To explore the effect of Schisandrae Sphenantherae on proliferation and differentiation of rat osteoblast in vitro. Methods Using the method of serum pharmacology, osteoblast was isolated from calvaria of newborn SD rats by means of modified sequential collagenase digestion and incubated in RPMI 1640 medium. The cell morphology was observed under a phase contrast inverted microscope. MTT assay, ALP activity, mineral node count were detected to determine the status and activities of proliferation and differentiation. Results In 10%, 15% concentration groups after cultured 48 h and 5%, 10% after cultured 72 h, 75% ethanol extracts of Schisandrae Sphenantherae significantly stimulated the proliferation (P