Effect of oxymatrine on the replication cycle of hepatitis B virus in vitro

AIM:To determine the antiviral mechanism or target of oxymatrine against hepatitis B virus(HBV).METHODS:HepG2.2.15 cells were incubated with culture medium containing 500 μg/mL of oxymatrine for 2 and 5 d.The surface antigen of HBV(HBsAg) and e antigen of HBV(HBeAg) in supernatant were determined by ELISA.HBV DNA in supernatant,and intracellular covalently closed circular DNA(cccDNA),relaxed circular DNA(rcDNA) and pregenomic RNA(pgRNA) were quantif ied by specif ic real-time polymerase chain reaction(PCR) or reverse transcription(RT)-PCR.RESULTS:Treatment with oxymatrine for 2 d and 5 d reduced the production of HBV by the cell line,as indicated by the decline of HBsAg(22.67%,t = 5.439,P = 0.0322 and 22.39%,t = 5.376,P = 0.0329,respectively),HBeAg(55.34%,t = 9.859,P = 0.0101 and 43.97%,t = 14.080,P = 0.0050) and HBV DNA(40.75%,t = 4.570,P = 0.0447 and 75.32%,t = 14.460,P = 0.0047) in the supernatant.Intracellular cccDNA was also markedly reduced by 63.98%(t = 6.152,P = 0.0254) and 80.83%(t = 10.270,P = 0.0093),and intracellular rcDNA by 34.35%(t = 4.776,P = 0.0413) and 39.24%(t = 10.050,P = 0.0097).In contrast,intracellular pgRNA increased by 6.90-fold(t = 8.941,P = 0.0123) and 3.18-fold(t = 7.432,P = 0.0176) after 500 μg/mL of oxymatrine treatment for 2 d and 5 d,respectively.CONCLUSION:Oxymatrine may inhibit the replication of HBV by interfering with the process of packaging pgRNA into the nucleocapsid,or inhibiting the activity of the viral DNA polymerase.

Inhibition of hepatitis B virus by oxymatrine in vivo

AIM To investigate the anti-HBV effect ofoxymatrine (oxy) in vivo.METHODS HBV transgenic mice were producedby micro-injection of a 4.2kb fragmentcontaining the complete HBV genomes.Expression level of HBsAg and HBcAg in thetransgenic mice liver was determined byimmunohistochemical assay.RESULTS Four groups (6 mice in each group)were injected intraperitoneally with oxy at thedosage of 100,200, and 300 mg/kg or with salineonce a day for 30 days. Both HBsAg and HBcAgwere positive in livers of all the six mice in thecontrol group (injected with saline), and werepositive in livers of two mice in 100 mg/kg groupand 300mg/kg group. In 200mg/kg group,HBsAg and HBcAg were negative in livers of allthe six mice. Based on the results, 200 mg/kg isthe ideal dosage to explore the effect of oxy atdifferent time points. According to the oxytreatment time, mice were divided into fourgroups: 10 d, 20 d, 30 d and 60 d (4 mice in eachgroup). Each mouse underwent liver biopsy twoweeks before the treatment of oxy. Down-regulation of HBsAg and HBcAg appeared aftertreatment of oxymatrine for 10 d and 20 d, Dane-like particles disappeared after the treatment ofoxy for 20d under electron microscopy,however, the expression level of HBsAg andHBcAg returned to normal 60 d later after oxytreatment.CONCLUSION oxymatrine can reduce thecontents of HBsAg and HBcAg in transgenic miceliver, longer treatment time and larger dosagedo not yield better effects.

Inhibitory effect of oxymatrine on hepatocyte apoptosis via TLR4/PI3K/Akt/GSK-3β signaling pathway

AIM To evaluate the effect of oxymatrine(OMT) on hepatocyte apoptosis in rats with lipopolysaccharide(LPS)/D-galactosamine(D-Gal N)-induced acute liver failure(ALF). METHODS LPS/D-Gal N was used to establish a model of ALF in rats. To evaluate the effect of OMT, we assessed apoptosis by transmission electron microscopy, and the pathological changes in the liver by light microscopy with hematoxylin and eosin staining. An automated biochemical analyzer was used to measure serum alanine aminotransferase(ALT) and aspartate aminotransferase(AST). Enzyme-linked immunosorbent assay was used to determine the levels of tumor necrosis factor(TNF)-α and interleukin(IL)-1β. Western blotting was used to detect protein levels in liver tissues. Streptavidin peroxidase immunohistochemistry was used to observe expression of Toll-like receptor(TLR)4, active caspase-3, Bax and Bcl-2. RESULTS All rats in the normal control and OMT-pretreated groups survived. The mortality rate in the model group was 30%. OMT preconditioning down-regulated apoptosis of hepatocytes and ameliorated pathological changes in liver tissue. The levels of AST, ALT, TNF-α and IL-1β in the model group increased significantly, and were significantly reduced by OMT pretreatment. OMT pretreatment down-regulated expression of TLR4 and active caspase-3 and the Bax/Bcl-2 ratio, and upregulated expression of P-AktSer473(Akt phosphorylated at serine 473) and P-GSK3βSer9(glycogen synthase kinase 3β phosphorylated at serine 9) induced by LPS/D-Gal N. CONCLUSION OMT inhibits hepatocyte apoptosis by suppressing the TLR4/PI3K/Akt/GSK-3β signaling pathway, which suggests that OMT is an effective candidate for ameliorating acute liver failure.

Effect of oxymatrine on interferon-gamma and tumor necrosis factor-alpha serum levels in an experimental rat model of autoimmune encephalomyelitis

BACKGROUND:Studies have demonstrated that experimental autoimmune encephalomyelitis(EAE) onset correlates with increased interferon-γ(IFN-γ) and tumor necrosis factor-α(TNF-α) expression.Oxymatrine(OM) has been shown to inhibit autoimmune responses,but there are no reports showing that it could prevent the development of EAE.OBJECTIVE:To observe the effect of OM on serum levels of IFN-γ and TNF-α in a rat model of EAE.DESIGN,TIME AND SETTING:A randomized,controlled,animal study was performed at the Experimental Animal Center of Henan Academy of Chinese Medicine and at the Key Disciplines Laboratory Clinical Medicine of Henan Province between July and December 2008.MATERIALS:OM was purchased from Chia-tai Tianqing Pharmaceutical,China;complete Freund's adjuvant was purchased from Sigma,USA.METHODS:Forty female Wistar rats were randomly assigned to four groups:EAE model(M),low-dose OM treatment(OM-L),high-dose OM treatment(OM-H),and normal control(N,no immunization),with 10 rats in each group.EAE was established in the M,OM-L,and OM-H groups following immunization with Guinea pig spinal cord homogenate and complete Freund's adjuvant.The M and N groups were intraperitoneally injected with normal saline(6.7 mL/kg per day),the OM-L group received an intraperitoneal injection of OM(100 mg/kg per day),and the OM-H group received OM(150 mg/kg per day).MAIN OUTCOME MEASURES:At 16 days after immunization,the degree of histopathological changes in the spinal cord was assessed by hematoxylin-eosin stanining.Enzyme-linked immunosorbent assay was used to detect serum levels of IFN-γ,and radioimmunoassay was utilized to determine serum TNF-α level.Neurological scores were measured on a daily basis according to a 0-5 scale.RESULTS:Daily injections of OM,both high and low doses,resulted in decreased neurological scores in EAE rats(P < 0.01),as well as reduced cellular infiltration in the spinal cord and decreased levels of serum IFN-γ and TNF-α(P < 0.01).CONCLUSION:OM reduced the onset and severity of EAE,which correlated with decreased IFN-γ and TNF-α expression.

Evaluation of pharmacokinetics underlies the collaborated usage of lamivudine and oxymatrine in beagle dogs

Combinational therapy of lamivudine and oxymatrine has been employed in the battle against hepatitis B virus in clinical setting. However, the pharmacokinetic behavior of the drug or active metabolism in intravenous/oral co-administration regime is poorly investigated. Herein,we evaluated the pharmacokinetic characteristic through a tailor-designed 3 way crossoverLatin square experiment in adult male beagle dogs. Six dogs were randomly treated by intravenous administration of lamivudine(2.5 mg/kg), oxymatrine(15 mg/kg) and combinational dosage, named as intravenous regime. Meanwhile the other six dogs were orally administrated with lamivudine(2.5 mg/kg), oxymatrine(15 mg/kg) and combinational dosage,named as oral regime. The pharmacokinetic feature in simultaneous oral treatment appeared to have no significant difference when compared with individual administration,even including matrine, the active metabolite of oxymatrine. In intravenous regime, the main pharmacokinetic parameters of simultaneous administration were nearly consistent with intravenous regime remedy. The collaborated application of lamivudine and oxymatrine contributed to non-distinctive pharmacokinetic fluctuations of beagle dogs in intravenous/oral regime, compared with individual employment, which established a vital base for the clinical co-administration against hepatitis B. Furthermore, the present study demonstrated that the determination of pharmacokinetics between combinational and individual therapy might assist in the development of drug compatibility in clinical therapy.

Differentially expressed genes identified by microarray analysis following oxymatrine treatment of hepatic stellate cell

AIM: To investigate the effects of oxymatrine on the gene expression profile of hepatic stellate cell (HSC) and provide novel insights into the mechanism of oxymatrine against hepatic fibrosis. Methods: HSC was isolated from normal SD by in situ perfusion of collagenase and pronase and density Nycodenz gradient centrifugation. MTT colorimetry was used to study the effect of oxymatrine on the proliferation of HSC. Total RNA and mRNA of quiescent HSC, culture-activated HSC and oxymatrine treated HSC were extracted. Effect of oxymatrine on HSC gene expression profile was detected by oligonucleotide microarray analysis with Affymetrix gene chip rat U230A. Differentially expressed genes were annotated with Gene Ontology (GO) and analyzed with Kyoto encyclopedia of genes and genomes (KEGG) pathway using the Database for Annotation, Visualization and Integrated Discovery. Results: Oxymatrine could inhibit the proliferation of HSC in a dose-dependent manner. A total of 4641 differentially expressed genes were identified by cDNA chip between activated and quiescent HSC, among which 2702 genes were upregulated, and 1939 genes were down-regulated in activated HSC. cDNA microarray uncovered downregulation of 56 genes in response to oxymatrine, the representative genes including alpha 2 type I procollagen, alpha-1 type I collagen, tissue inhibitor of metalloproteinase 1, interleukin 1 beta, early growth response 1, chemokine ligand 2, chemokine ligand 1, CTGF, TGFβ1. The most enriched GO terms included response to wounding, inflammatory response, cell migration, cell motility, wound healing, TGFβ receptor signaling pathway. KEGG pathway analysis revealed that oxymatrine affected the ECM-receptor interaction, focal adhesion, cytokine-cytokine recaptor interaction, TGFβ signaling pathway, MAPK signaling pathway. There were 37 genes upregulated significantly following oxymatrine treatment. The most enriched GO terms included oxidation reduction, negative regulation of lipoprotein oxidation, regulation of lipoprotein oxidation, steroid metabolic process, regulation of lipase activity. Six genes were confirmed with QPCR, consistent with microarray. Conclusion: The mechanism of oxymatrine in inhibiting liver fibrogenesis is associated with multi-genes and multi-pathways regulation.

Effects of oxymatrine on calcium channels and GABA release in mice under neuropathic pain condition

OBJECTIVE To investigate effects of oxymatrine,an alkaloid from Sophora flavescens Ait.,on high-voltage dependent calcium channel and inhibitory neurotransmitter GABA under neuropathic pain condition.METHODS The partial sciatic nerve ligation(PSNL)was executed on C57/BL6 mice to produce neuropathic pain.Oxymatrine(150 mg·kg-1)was administrated intraperitoneally to PSNL mice.Mechanical hindpaw withdral threshold(MWT)was measured under Von-Frey filament stimulation with up-and-down method.In brain tissue,GABA concentration was measured with ELISA.Change of GABAAreceptor protein expression,N-type calcium channel(Cav2.2)and L-type calcium channel(Cav1.3)protein expressions were detected with Western-blot;intracellular calcium concentration was measured in cultured cortical neurons with Fluo-3/AM fluorescent probe.RESULTS Compared to saline,oxymatrine significantly increased ED50 of MWT on PSNL mice(P<0.05).GABA concentration and GABAAreceptor protein level in brain tissue were decreased in PSNL mice,while administration of oxymatrine increased both GABA concentration and GABAA receptor expression.Intracellular calcium concentration was increased in cultured cortical neurons by oxymatrine treatment,but this phenomenon was not seen under calcium-free condition.Protein expression of Cav2.2,but not Cav1.3,was found to be decreased in the brains of PSNL mice and to be restored to a normal level with oxymatrine administration.CONCLUSION Oxymatrine has analgesic effect on PSNL-induced neuropathic pain in mice.This phenominon relates to the increase of GABA release,GABAAreceptor expression,and also the restoration of expression level of Cav2.2 but not Cav1.3 in brain tissues,which suggesting that Ca2+ flow through Cav2.2 calcium channel may be the key point underlying oxymatrine analgesia.

Simultaneous determination of berberine,matrine and oxymatrine in traditional Chinese medicines by using nonaqueous capillary electrophoresis

A rapid method for the simultaneous determination of berberine(BBR),matrine(MT)and oxymatrine(OMT)by nonaqueous capillary electrophoresis(NACE)was developed.Optimum separation of the analytes was obtained on a 50cm×50μm i.d.fused-silica capillary using a non-aqueous buffer system of 70mM ammonium acetate,7.0% acetic acid and 10% acetonitrile at 25kV and 20℃.The relative standard deviations(R.S.D.)of the migration times and peak areas of the three active components were 0.06%-0.20% and 0.12%-3.41% for berberine,0.11%-0.60% and 0.74%-1.63% for matrine,0.15% and 0.45% for oxymatrine,respectively.Detection limits of berberine,matrine and oxymtrine were 0.18μg/mL,4.08μg/mL and 4.16μg/mL,respectively.In the tested concentration range,good linear relationships(0.9992 for berberine,0.9988 for matrine and 0.9988 for oxymatrine)were observed.The linear calibration ranges were 0.45-360.0μg/mL for berberine,8.16-408.0μg/mL for matrine and 20.8-416.0μg/mL for oxymatrine.This method has been successfully applied to the phytochemical analysis of alkaloids extracts from two commonly used traditional Chinese herbal drugs:Sophora flavescens Ait.(Kushen)and Cortex phellodendri chinensis(Huangbai)and their medicinal preparations.

Effects and Mechanism of Oxymatrine on Proliferation,Invasion and Migration of Hepatocellular Carcinoma Cells

[Objectives] To observe the effects of oxymatrine on the proliferation,invasion and migration of hepatocellular carcinoma cells,and to explore its possible molecular mechanism.[Methods]The hepatocellular carcinoma cell line Hep G2 was randomly divided into the negative control group and the low,medium and high concentration groups of oxymatrine.The negative control group was added to the cell culture medium,and the low,medium and high concentration groups of oxymatrine were added to the oxymatrine and cell culture medium with the final concentration of 1.0,2.0 and 4.0 mg/m L.The proliferation of each group was measured by MTT assay,and the inhibition rate of cell proliferation was calculated.The invasion and migration ability of each group was determined using Transwell chamber assay.The expression of E-cadherin and CD44 mRNA in each group was detected using Real-time PCR,while the expression of E-cadherin and CD44 protein was detected using Western blotting method.[Results] The inhibition rate of cell proliferation in the high concentration group of oxymatrine was higher than that in the medium and low concentration groups,and the inhibition rate of cell proliferation of medium concentration group was higher than that in the low concentration group.In the cell invasion and cell migration experiments,the number of transmembrane cells of low,medium and high concentration groups of oxymatrine was less than that in the negative control group(P < 0.05),the number of transmembrane cells of high concentration groups of oxymatrine was less than that in medium and low concentration groups,and the number of transmembrane cells of medium concentration group was less than in that of the low concentration group(P < 0.05).Compared with the negative control group,the expression of E-cadherin mRNA and protein increased in the low,medium and high concentration groups of oxymatrine,while the expression of CD44 mRNA and protein decreased(P < 0.05).[Conclusions] Oxymatrine has the effect of inhibiting the proliferation,invasion and migration of hepatocellular carcinoma cell line Hep G2 in vitro.The higher the concentration,the more significant the effect will be.The mechanism is possibly connected with the up-regulation of E-cadherin expression and down-regulation of CD44 expression.

Quantitative Analysis of Matrine and Oxymatrine in Sophora flavescens Extract and Its Biopesticides by UPLC

Successive cartridge clean-up method for the simultaneous determination of matrine and oxyma- trine in biopesticides containing Sophora flavescens extract was developed and validated by UPLC. The clean-up method was established with ENVI-Carb (0.5 g) and C18 SPE (0.5 g) cartridges for the bioactive alkaloid in biopesticides from S. flavescens, and the eluate was analyzed to quantify the matrine and oxymatrine by UPLC. The developed method was validated, and the recovery and LOQ of both materials were 105.0% and 103.6%, and 0.050 and 0.684 mg·kg-1, respectively. Of the twenty one samples, the total content of matrines were analyzed by using the developed method and the result showed the developed successive clean-up method could contribute to the manufacture and control of biopesticides including matrines, and can be ap- plied to the method development for the analysis of alkaloid materials in biopesticides.

T cell–associated immunoregulation and antiviral effect of oxymatrine in hydrodynamic injection HBV mouse model

Although oxymatrine(OMT) has been shown to directly inhibit the replication of hepatitis B virus(HBV) in vitro, limited research has been done with this drug in vivo. In the present study, the antiviral effect of OMT was investigated in an immunocompetent mouse model of chronic HBV infection.The infection was achieved by tail vein injection of a large volume of DNA solution. OMT(2.2, 6.7 and20 mg/kg) was administered by daily intraperitoneal injection for 6 weeks. The efficacy of OMT was evaluated by the levels of HBV DNA, hepatitis B surface antigen(HBs Ag), hepatitis B e antigen(HBe Ag)and hepatitis B core antigen(HBc Ag). The immunoregulatory activity of OMT was evaluated by serum ELISA and flow cytometry. Results shows that OMT at 20 mg/kg inhibited HBV replication, and it was more efficient than entecavir(ETV) in the elimination of serum HBs Ag and intrahepatic HBc Ag. Inaddition, OMT accelerated the production of interferon-γ(IFN-γ) in a dose-dependent manner in CD4^+T cells. Our findings demonstrate the beneficial effects of OMT on the enhancement of immunological function and in the control of HBV antigens. The findings suggest this drug to be a good antiviral therapeutic candidate for the treatment of HBV infection.

Anti-asthmatic effects of oxymatrine in a mouse model of allergic asthma through regulating CD40 signaling

The aim of the study was to investigate the anti-asthmatic effects of oxymatrine(OXY) and the possible underlying mechanisms.The mouse asthma model was established by ovalbumin(OVA) intraperitoneal injection.A total of fifty mice were randomly assigned to five groups:control,OVA,OVA + dexamethasone(Dex,2 mg·kg-1),and OVA + OXY(40 mg·kg-1),and OVA + OXY(80 mg·kg-1),respectively.Histological studies were conducted by the hematoxylin and eosin(HE) staining,the levels of interleukin-4(IL-4),interleukin-5(IL-5),interleukin-13,and Ig E were evaluated by enzyme-linked immunosorbent assay(ELISA),and the protein level of CD40 was analyzed by Western blotting.OXY inhibited OVA-induced increases in eosinophil count;the levels of IL-4,IL-5,Ig E,and IL-13 were recovered.It also substantially inhibited OVA-induced eosinophilia in lung tissues and the expression of CD40 protein.These findings suggest that OXY may effectively ameliorate the progression of asthma and could be explored as a possible therapy for patients with allergic asthma.

Oxymatrine Inhibits Bocavirus MVC Replication, Reduces Viral Gene Expression and Decreases Apoptosis Induced by Viral Infection

Oxymatrine(OMT), as the main active component of Sophoraflavescens, exhibits a variety of pharmacological properties,including anti-oxidative, anti-inflammatory, anti-tumor, and anti-viral activities, and currently is extensively employed to treat viral hepatitis; however, its effects on parvovirus infection have yet to be reported. In the present study, we investigated the effects of OMT on cell viability, virus DNA replication, viral gene expression, cell cycle, and apoptosis in Walter Reed canine cells/3873 D infected with minute virus of canines(MVC). OMT, at concentrations below 4 mmol/L(no cellular toxicity), was found to inhibit MVC DNA replication and reduce viral gene expression at both mRNA and protein levels, which was associated with the inhibition of cell cycle S-phase arrest in early-stage of MVC infection.Furthermore, OMT significantly increased cell viability, decreased MVC-infected cell apoptosis, and reduced the expression of activated caspase 3. Our results suggest that OMT has potential application in combating parvovirus infection.

Preliminary Study on Efficacy of Oxymatrine in Treatment of Patients with Chronic Hepatitis C

Objective: To evaluate the efficacy of oxymatrine in the treatment of chronic hepatitis C and todiscuss its mechanism. Methods: Forty-three patients with chronic HCV infection were randomly divided intothe treated group (20 cases) and the control group (23 cases). The treated group was given oxymatrine 600 mgper day intramuscularly for three months, and the control group was given the general liver protective agentssuch as vitamins. Serum HCV-RNA, alanine aminotransferase (ALT), soluble interleukin--2 receptor and collagen type tV (IV-C) were determined before and after treatment. Results: Eight out of 17 HCV-RNA-positive(47. 1 % ) in the treated group converted to HCV-RNA-negative cases, while in 18 cases of the control group,the negative convertion only took place in 1 patient (5. 6% ), the negative conversion rate was significantlyhigher in the treated group than that in the control group (P < 0. 05). The normalization rates of serum ALT ofthe treated group at the end of the first and second month treatment were higher than those of the controlgroup, but after three months treatment, the normalization rates of the two groups were not different significantly. BOth serum levels of IV-C and plasma levels of soluble interleukin-2 receptor were significantly reducedafter oxymatrine treatment for three months (P < 0. 05). Conclusion: Oxymatrine is effective on eliminatingHCV-RNA and reducing fibrosis activity, so it could be a safe, effective drug in the treatment of chronic hepatitis C.Keywods: oxymatrine; hepatitis C virus ribonucleic acid; soluble interleukin-2 receptor; collagen

Oxymatrine could promote mesenchymal stem cell therapy in hepatic fibrosis rats:an experimental research

Objective To investigate whether oxymatrine (OM) could promote mesenchymal stem cell (MSC) therapy in CCl4-induced hepatic fibrosis (HF) in rats and to initially explore its mechanisms.Methods Totally 50 male SD rats were randomly divided into five groups,i.e.,nor-

Quality Standard of Qiang Herb Sophora davidii var.chuansiensis

[Objectives]To establish the quality standard of common clinical folk herb Sophora davidii var.chuansiensis.[Methods]The qualitative identification was performed by microscopic method and thin-layer chromatography(TLC).The contents of moisture,total ash,acid insoluble ash and extract were determined according to the methods of the Chinese Pharmacopoeia(2020).High performance liquid chromatography(HPLC)was used to determine the contents of oxymatrine and oxysophocarpine.[Results]The microscopic characteristics were obvious,including crystal sheath fiber,calcium oxalate square crystal,non glandular hair,stone cells,epidermal cells,stomata,cork cells,vessels with marginal pits and so on.TLC spots were clear,and the resolution was good.The contents of moisture,total ash,acid insoluble ash and extract from 10 batches of samples were 4.70%-8.33%,3.43%-4.19%,0.65%-1.02%and 14.67%-22.04%respectively.The determination results of oxymatrine were between 0.10%-0.33%,with an average value of 0.19%.The determination results of sophocarpine oxide were between 0.30%-0.38%,with an average value of 0.34%.[Conclusions]The established quality standard of S.davidii var.chuansiensis had good specificity and accuracy,and could be used for the quality control of herb S.davidii var.chuansiensis.

氧化苦参碱注射液对大鼠实验性脑出血后作用的研究

氧化苦参碱（oxymatrine,OMT）是从豆科槐属植物苦参（So-phora flavescensAi.t）等中提取的生物碱,分子式为C15H24N2O,具有四环的喹嗪啶类结构。研究发现,OMT延缓乌头碱所致大鼠心律失常的发生,对TLR4表达有抑制作用。降低心肌细胞凋亡指数,抑制Fas蛋白表达,增强Bcl-2蛋白表达。

Preparation and Evaluation of MPEG-PCL Polymeric Nanoparticles Against Gastric Cancer

This paper provides a new treatment for gastric cancer with a new OMT delivery system.We synthesized MPEG-PCL,an amphiphilic polymer,to construct a nanoparticle encapsulated OMT by pH gradient method,and then examined the nanodrug’s therapeutic efficacy.An integral analytical method was used to characterize the structure of MPEG-PCL.The single factor method and orthogonal test were utilized to investigate the optimum preparation process.The morphology and average size of the OMT-NPs were analyzed by transmission electron microscopy and Zetasizer.CCK-8 assay and confocal fluorescent microscope were used to study the inhibitory effect on SGC-7901 gastric cancer cells.The average size of nanoparticles was 95.86±1.54 nm.The maximum encapsulation efficiency of OMT was 46.84±4.37%,while the drug loading content was 8.89±1.09%.The cumulative release of nanoparticles was 73.07±1.5%,inspected through dynamic dialysis in vitro.Compared with free OMT,OMT-NPs showed enhanced cytotoxic effects in SGC-7901 cells.The nanoparticles could efficiently deliver the OMT into the cancer cells and release it.The OMT delivery system prepared in this paper provides a potential platform for the treatment of gastric cancer..

Study on possible synergistic anticancer cachexia effects of the main active compounds of Radix Sophorae flavescentis

Background:Cancer cachexia is a complex disease secondary to cancer,and no specific therapy for it has been found.The Chinese herb Kushen(Radix Sophorae flavescentis)is the dried root of Sophora flavescens Aiton,which has been widely applied in treating digestive and urinary inflammatory diseases.Matrine,one of the main components of Radix Sophorae flavescentis,can alleviate cancer cachexia.Methods:Compounds from Radix Sophorae flavescentis were screened using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform.The targets related to cancer cachexia were queried from the Therapeutic Target Database(http://db.idrblab.net),DisGeNET database(http://www.disgenet.org),and Search Tool for Interacting Chemicals database,related literature,and constructed cancer cachexia-protein network.Cancer cachexia-protein network was merged with compound-protein networks respectively using Cytoscape software as well as network topology data and key targets counting.Pathway enrichment analysis was conducted via the Database for Functional Annotation Bioinformatics Microarray Analysis.Protein crystal structures and compound structures were queried from RCSB and PubChem databases,respectively.Molecular docking was conducted using Discovery Studio software.Results:The anticancer cachexia compounds of Radix Sophorae flavescentis were screened as oxymatrine,matrine,and kurarinol,and targets such as BIRC2,TNF and STAT3 were found.The mechanisms of oxymatrine,matrine,and kurarinol have the characteristics of synergy and complementarity.Kurarinol has a mechanism similar to that of matrine,which includes the FoxO signaling pathway,insulin resistance and mTOR signaling pathway.TNF signaling pathway is a common signaling pathway of kurarinol,oxymatrine and matrine.Adipocytokine signaling pathway is the other common pathway of kurarinol and oxymatrine except for the TNF signaling pathway.Kurarinol can be successfully docked with CYCS,GPX2,BIRC7,etc.,and kushenol C can be successfully docked with IKBKB and PIK3CD.Conclusion:Kurarinol,matrine,oxymatrine,and kushenol C may be the key compounds of Radix Sophorae flavescentis treating cancer cachexia.Additionally,TNF signaling pathway is the key pathway for the synergistic action of kurarinol,matrine and oxymatrine.