Matrine promotes colorectal cancer apoptosis by downregulating shank-associated RH domain interactor expression

BACKGROUND The 5-year survival rate of patients with colorectal cancer(CRC)in China is only 56.9%,highlighting the need for new therapeutic drugs.Previous studies have shown that matrine exhibits antitumor effects by inducing apoptosis.However,the mechanism by which matrine regulates antiapoptotic proteins in CRC remains unclear.AIM To identify apoptotic proteins from proteomics and investigate the role of matrine in impeding CRC apoptosis by regulating these proteins.METHODS Tumor and adjacent normal tissues were collected from 52 patients with CRC who underwent surgery between January and December 2021.Data-independent acquisition quantitative proteomic analysis was performed to identify differentially expressed apoptotic proteins.The selected apoptotic proteins were identified through their association with tumor-node-metastasis(TNM)stage and prognosis,then confirmed by immunohistochemical(IHC)staining in validation cohort.In vitro,the role of matrine or apoptotic proteins on cancer cells were analyzed.RESULTS Compared to normal tissues,88 anti-apoptotic proteins from proteomic results were selected.Among them,Shankassociated RH domain interactor(SHARPIN)was identified because of its relationship with TNM stage and overall survival in TCGA database.In the IHC-confirmed cohort,SHARPIN was highly expressed in CRC tissues and localized in the cytoplasm.Higher SHARPIN expression was associated with TNM stage,carbohydrate antigen 153 levels,and gross type compared to low expression.SHARPIN knockdown promoted apoptosis,significantly upregulated the expression of Bcl-2 associated agonist of cell death,Bcl-2 associated X protein,caspase 3,and caspase 8,and downregulated B-cell lymphoma-2(P<0.05).Importantly,matrine treatment promoted apoptosis and reversed the proliferation,invasion,and migration of CRC cells by repressing SHARPIN.CONCLUSION SHARPIN was identified as an upregulated anti-apoptotic protein in CRC,and matrine exhibited anticancer effects by downregulating its expression.Thus,matrine appears to be a promising drug for CRC.

Study on the mechanism of the anti-tumor effects of matrine via regulation of lipid metabolism mediated by SREBP signaling pathway

Objective:To investigate the therapeutic effects of matrine on CT26 tumor-bearing mice and its effect on the sterol regulatory element binding protein(SREBP)signaling pathway.Methods:A CT26 tumor-bearing mouse model was established using CT26 cells.Different doses of matrine were orally administered to mice,and the tumor size and weight in each group of mice before and after administration were measured to calculate the tumor inhibition rate of matrine.Subsequently,tumor tissues were subjected to hematoxylin and eosin(HE)staining to observe morphological changes in tumor tissue,and quantitative polymerase chain reaction(qPCR)was performed to detect the expression of the genes of the lipid metabolism-related enzymes sterol regulatory element binding transcription factor 1(Srebf1),ATP citrate lyase(Acly),acetyl-Coenzyme A carboxylase alpha(Acc),and fatty acid synthase(Fasn)in tumor tissues before and after matrine intervention.Results:Compared to those in the model group,tumor-bearing mice in both the low and high-dose matrine groups showed significantly reduced tumor weights.HE staining showed that matrine significantly inhibited tumor cell proliferation in both the low and high-dose groups.The qPCR results showed that,compared with the model group,the expression levels of the genes of lipid metabolism-related enzymes Srebf1,Acly,Acc,and Fasn in tumor tissues were significantly downregulated in both the low and high-dose matrine groups.Conclusion:Matrine modulates the lipid metabolism pathway,affects tumor cell lipid metabolism,and exerts antitumor effects.

Effects of Matrine on Adenine-Induced Chronic Tubulointerstitial Fibrosis in Rats

Aim ,To study the mechanism relative to therapeutic effects of matrine on adenine-induced renal interstitial fibrosis in rats. Methods Sixty male Wistar rats were selected and randomly divided into 3 groups： normal control group （NCG） consisted of 8 rats, adenine treated group （ATG） 28 rats, and matrine treated group （MTG） 24 rats. Each rat in ATG and MTG was gavaged with adenine （250 mg·kg^-1·d^-1 ） for 21 d. After gavage with adenine for one week, each rat in MTG was administered intraperitoneally matrine（20 mg·kg^-1·d^-1 ） in vehicle （ 1 mL of 0.9% sodium chloride） daily. On days 14, 21, and 28, the serum levels of urea nitrogen, creatinine, and IL-6 were determined and the rat kidneys of ATG, MTG and NCGwere examined pathologically. Radioimmunoassay for serum IL- 6 immunohistochemical staining for TGF-β1 expression in the kidney and semiquantitative analysis were performed. HE staining for semiquantitative analysis of tubulointerstitial injury. Results The serum levels of urea nitrogen and creatinine in MTG were lower as compare to ATG （ P 〈 0.05 ） whereas serum IL- 6 and renal TGF-β1 expression levels were significantly lower than those in ATG （P 〈0.05 ）, but all these indexes were higher than those in NCG （P 〈 0.01 ）. In MTG, the index of tubulointerstitial lesion was lower than that in ATG （P 〈 0.05 ）. Conclusion Matrine inhibits the renal tubulointerstial fibrosis in the adenine-induced rat model by suppressing serum level of IL-6 and expression of TGF-β1 in the tubulointerstitium.

Antifungal activities of matrine and oxymatrine and their synergetic effects with chlorthalonil

The EC50 values of matrine and oxymatrine against five forest pathogenic fungi （Fusarium oxysporum, Valsa pini, Cladosporium oxysporum, Sphaeropsis sapinea, Marssonina brunnea） were examined by bioassay methods. The results demonstrated that matrine and oxymatrine had strong inhibitory activities to the conidium germination of the tested fungi. The ECso values of matrine for inhibiting the conidium germination of Marssonina brunnea, Cladosporium oxysporum, Sphaeropsis sapinea were 123μg·mL^-1, 272 μg·mL^-1, 1133 μg·mL^-1, respectively, and the EC50 values of oxymatrine for inhibiting the conidium germination of Fusarium oxysporum, Sphaeropsis sapinea were 532μg·mL^-1, 601μg·mL^-1, respectively. The hyphal growth of the fungi was also significantly inhibited by matrine and oxymatrine. The ECs0 values of matrine inhibiting the conidium germination of Sphaeropsis sapinea, Valsa pini, Fusarium oxysporum were 428μg·mL^-1, 535 μg·mL^-1, 592 μg·mL^-1, respectively. The EC50 values of oxymatrine inhibiting the conidium germination of Valsa pini, Fusarium oxysporum were 323, 618μg·mL^-1, respectively. In the synergetic tests the ECs0 values of the mixtures of thiophanate methyl （or chlorthalonil） and matrine （or oxymatrine） were lower than 34 μg·mL^-1 while their co-toxicity coefficients were significantly higher than 1130 It indicated that the mixture of the alkaloids and the chemical had potential practical utilization in controlling certain forest fungal diseases.

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.

Determination of Matrine and Oxymatrine in Radix Sophorae Flavescentis by Resonance Rayleigh Scattering, Second-order Scattering and Frequency Doubling Scattering Technique

In 0.1 mol/L HCl medium, 12-tungstophosphoric（TP） acid reacted with matrine（Mat） and oxymatrine（Oxy） to form an ion-association complex. As a result, the new spectra of resonance Rayleigh scattering（RRS）, second-order scattering（SOS） and frequency doubling scattering（FDS） appeared and their intensities were enhanced greatly. The maximum scattering wavelengths of RRS, SOS and FDS were located at 370, 670 and 390 nm, respectively. The in-crements of scattering intensity were directly proportional to the concentration of Mat and Oxy in a certain range. Based on this, the method for the determination of matrine and oxymatrine has been established. It has been applied to the determination of matrine and oxymatrine in samples of Radix sophorae flavescentis with satisfactory result. The reaction mechanism and reasons of RRS enhancement were discussed.

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.

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.

Antitumor effect of matrine in human hepatoma G2 cells by inducing apoptosis and autophagy

AIM: To study the antitumor effect of matrine in human hepatoma G2 (HepG2) cells and its molecular mechanism involved in antineoplastic activities. METHODS: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to detect viability of HepG2 cells. The effect of matrine on cell cycle was detected by flow cytometry. Annexin-V-FITC/PI double staining assay was used to detect cellular apoptosis. Cellular morphological changes were observed under an inverted phase contrast microscope. Transmission electron microscopy was performed to further examine ultrastructural structure of the cells treatedwith matrine. Monodansylcadaverine (MDC) staining was used to detect autophagy. Whether autophagy is blocked by 3-methyladenine (3-MA), an autophagy inhibitor, was evaluated. Expression levels of Bax and Beclin 1 in HepG2 cells were measured by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR).RESULTS: Matrine signif icantly inhibited the proliferation of HepG2 cells in a dose- and time-dependent manner, and induced G1-phase cell cycle arrest and apoptosis of HepG2 cells in a dose-dependent manner. The total apoptosis rate was 0.14% for HepG2 cells not treated with matrine. In contrast, the apoptosis rate was 28.91%, 34.36% and 38.80%, respectively, for HepG2 cells treated with matrine at the concentration of 0.5, 1.0 and 2.0 mg/mL. The remarkable morphological changes were observed under an inverted phase contrast microscope. Abundant cytoplasmic vacuoles with varying sizes were observed in HepG2 cells treated with matrine. Furthermore, vacuolization in cytoplasm progressively became larger and denser when the concentration of matrine was increased. Electron microscopy demonstrated formation of abundant autophagic vacuoles in HepG2 cells after matrine treatment. When the specif ic autophagic inhibitor, 3-MA, was applied, the number of autophagic vacuoles greatly decreased. MDC staining showed that the fluorescent density was higher and the number of MDC-labeled particles in HepG2 cells was greater in matrine treatment group than in control group. Fewer autophagic vacuoles were observed in the combined 3-MA and matrine treatment group when 3-MA was added before matrine treatment, indicating that both autophagy and apoptosis are activated when matrine-induced death of hepatoma G2 cells occurs. Real-time quantitative RT-PCR revealed that the expression levels of Bax gene, an apoptosis-related molecule, and Beclin 1 gene which plays a key role in autophagy were higher in matrine treatment group than in control group, indicating that Beclin 1 is involved in matrineinduced autophagy and the pro-apoptotic mechanismof matrine may be related to its upregulation of Bax expression. CONCLUSION: Matrine has potent antitumor activities in HepG2 cells and may be used as a novel effective reagent in treatment of hepatocellular carcinoma.

Effects of Matrine on Aconitine-Induced Electrophysiological Changes in Rat Ventricular Myocytes

Aim To explore the reason that the antiarrhythmic effect of the extract oftraditional Chinese medicinal herb, matrine, is weaker than quinidine and verapamil by comparison ofthe effect and efficacy of matrine on various kinds of transmembrane ionic currents with those ofquinidine and verapamil; and to demonstrate the best targets for antiarrhythmic drugs. MethodsWhole-cell patch-clamp techniques were used to record the action potential and ionic currents insingle cells of rat ventricular myocytes. Aconitine was used to induce the changes of ioniccurrents, then study the effects of matrine and quinidine, verapamil on aconitine-induced unbalancedchannel currents and action potential. Results Aconitine 1 μmol·L^(-1) induced significantchanges in transmembrane currents and action potential in single cells of rat ventricular myocytes.APD was significantly prolonged by aconitine. Simultaneously, aconitine increased sodium, L-typecalcium and inward rectifier potassium currents. Matrine 100 μmol· L^(-1) reversed theaconitine-induced changes of sodium current (I_(Na)) from (-70.2+- 10.5) pA/pF to ( - 39.6+-4.0)pA/pF(n = 5, P < 0.05 vs aconitine); L-type calcium current (I_(Ca-L)) from (20.4+- 3.8) pA/pF to (- 12.9+- 2.9) pA/pF ( n = 6, P < 0.01); the inward rectifier potassium current (I_(k1) ) from (-32.2+- 1.08) pA/pF to ( -24.0+-3.4) pA/pF (n = 6, P < 0.01), and action potential duration. Thereversal effects of quinidine and verapamil on aconitine-induced changes of APD and ionic currentswere more marked than matrine. Conclusion Aco-nitine significantly disturbs the normal equilibriumof ion channels in ventricular myocytes. It induces changes of I_(Na), I_(Ca-L), I_(K1) andprolongation of action potential duration. Matrine at concentration 50 or 100 μmol·L^(-1)statistically significantly suppresses aconitine-induced changes of APD and ionic currents. Thepotency and efficacy of inhibitory effect of matrine are markedly weaker than those of commonly usedverapamil and quinidine.

Synergistic protection of astragalus polysaccharides and matrine against ulcerative colitis and associated lung injury in rats

BACKGROUND Ulcerative colitis(UC)is a main form of inflammatory bowel disease.Due to complicated etiology and a high rate of recurrence,it is quite essential to elucidate the underlying mechanism of and search for effective therapeutic methods for UC.AIM To investigate the effects of astragalus polysaccharides(APS)combined with matrine on UC and associated lung injury.METHODS UC was induced in rats by colon mucosal tissue sensitization combined with trinitro-benzene-sulfonic acid-ethanol.Then,the effects of the treatments of salazopyrine,APS,matrine,and APS combined with matrine on histopathological changes of lung and colon tissues,disease activity index(DAI),colon mucosal damage index(CMDI),serum endotoxin(ET)level,serum diamine oxidase(DAO)activity,the contents of tumor necrosis factor-αand interleukin-1β,and the activities of myeloperoxidase,superoxide dismutase,and malondialdehyde in lung tissues,as well as the protein expression of zonula occludens(ZO)-1,Occludin,and trefoil factor 3(TFF3)were detected in UC rats.RESULTS The treatments of salazopyrine,APS,matrine,and APS combined with matrine reduced DAI scores and improved histopathological changes of colon and lung tissues,as well as decreased CMDI scores,ET levels,and DAO activities in UC rats.Moreover,in lung tissues,inflammatory response and oxidative stress injury were relieved after the treatments of salazopyrine,APS,matrine,and APS combined with matrine in UC rats.Furthermore,the expression of ZO-1,Occludin,and TFF3 in lung and colon tissues was increased after different treatments in UC rats.Notably,APS combined with matrine exerted a better protective effect against UC and lung injury compared with other treatments.CONCLUSION APS combined with matrine exert a synergistic protective effect against UC and lung injury,which might be associated with regulating TFF3 expression.

Efficacy of intramuscular matrine in the treatment of chronic hepatitis B

BACKGROUND: Hepatitis B virus (HBV) infection, a glo- bal public health problem, is the leading cause of cirrhosis and hepatocellular carcinoma (HCC) worldwide. There are more than 350 million HBV carriers in the world and up to one million die annually due to hepatitis B associated liv- er disease. So far no optimal treatment is available for pa- tients with chronic hepatitis B. In the paper we investigated the efficacy of intramuscular matrine in the treatment of chronic hepatitis B. METHODS: One hundred and twenty patients with chronic hepatitis B were randomly divided into matrine treatment group (n =60) and control group (n =60). The patients of the matrine group were given intramuscularly with matrine (an alkaloid extracted from a traditional Chinese herb Radix Sophorae Flavescentis by Guangzhou Ming Xing Pharmaceu cal Factory, Guangzhou, China) of 100 mg daily for 90 days in addition to conventional liver-protective drugs in- cluding glucurone, inosine, compound vitamin B and caryophyllin. The control group received conventional liv- er-protective drugs alone. Clinical manifestations and labo- ratory parameters including liver biochemistry and serum hepatitis B virus markers were monitored before and after treatment in the two groups. RESULTS: Significant differences were seen between the two groups in terms of improvement of clinical symptoms and signs, recovery of liver functions, and serum conver- sion from hepatitis Be antigen to HBe antibody and from positive to negative serum HBV DNA (P <0.05-0.01). The result of the matrine group was more marked than that of the control group. Serious side-effects were not observed except mild pain at the site of injection of matrine in a few patients. CONCLUSION: These results indicate that intramuscular matrine may be an economical, efficacious, safe drug for the treatment of chronic hepatitis B.

Matrine inhibits bladder cancer cell growth and invasion in vitro through PI3K/AKT signaling pathway:An experimental study

Objective:To study the inhibitory effect of matrine on bladder cancer cell growth and invasion in vitro through PI3K/AKT signaling pathway. Methods:Human T24 bladder cancer cell lines were cultured and treated with different doses of matrine(0.25 mg/mL,0.5 mg/mL and 1.0 mg/mL) as well as 20 μmol/L PI3K inhibitor LY294002 for 24 h,and the cell proliferation activity,the number of invasive cells as well as the expression of p-PI3K,p-AKT,proliferation genes and invasion genes were determined. Results:Different doses of matrine could decrease the cell viability value,the number of invasive cells as well as the expression of p-PI3K,p-AKT,MMP2 and MMP9,and increase the expression of p16,p21 and p27 in dose-dependent manner; p16,p21 and p27 expression in cells of 20 μmol/L LY29002 group were significantly higher than those of 0 μmol/L LY29002 group while MMP2 and MMP9 expression were significantly lower than those of 0 μmol/L LY29002 group(P<0.05). Conclusions:Matrine can inhibit bladder cancer cell proliferation and invasion in vitro and regulate the expression of cell cycle-inhibiting molecules and invasion-related genes through PI3K/AKT signaling pathway.

Anti-tumor effect of matrine combined with cisplatin on rat models of cervical cancer

Objective:To observe the anti-tumor effect of matrine combined with cisplatin on U14 rat models of cervical cancer.Methods:A total of 80 female Kunming rats were used to establish U14 rat models of cervical cancer and then divided into groups Ⅰ,Ⅱ,Ⅲ and Ⅳ,with 20 rats in each.For Group Ⅰ,the control group,injection of normal saline was given around the tumors.For Group Ⅱ,injection of 2 mg/kg cisplatin was given around the tumors.For Group Ⅲ,injection of 75 mg/kg matrine was given around the tumors while the combined injection of matrine and cisplatin was given for Group Ⅳ with the same doses as Groups Ⅱand Ⅲ.The animals were sacrificed 10 d after the injection and tumors were taken out for the comparisons of tumor weights after injection and calculation of anti-tumor rates,while thymus and spleen were taken for thymus index and spleen index.Blood in eyeball was collected for determination of changes in serum creatinine and urea nitrogen levels.Sections of tumor issue were prepared and morphological changes in tumor tissue cells were observed by using immunohistochemistry technique.Results:After injection,the thymus index and spleen index in Groups Ⅲ and Ⅳ were significantly higher than those in Groups Ⅰ and Ⅱ(P<0.05)while the two indexes in Group Ⅱ were significantly lower than Group Ⅰ(P<0.05).The tumor weights in Groups Ⅱ and Ⅳ were significantly smaller than those in Groups Ⅰ and Ⅲ(P<0.05) with significantly higher anti-tumor rates than Groups Ⅰ and Ⅲ(P<0.05).The serum creatinine and urea nitrogen levels in Groups Ⅲ and Ⅳ were significantly lower than Group Ⅱ(P<0.05) and the two indicators in Group Ⅲ were significantly lower than those in Group Ⅳ(P<0.05).The observation under the histological microscope showed densely arranged tumor cells in Group Ⅰ,growing as a crumby structure and diffuse appearance,with hyperchromatic and large nuclei,and abundant cytoplasm.In the case of Group Ⅱ,it showed less tumor cells,with extensive degenerative necrosis,sparse arrangement and karyopyknosis as well as karyoclasis.For Group Ⅲ,necrosis of tumor cells in different sizes and heterogeneous color in nuclei were observed.For Group Ⅳ,the number of tumor cells was significantly smaller than Groups Ⅰ and Ⅲ and the tumor cells presented an appearance of crumby structure as cancer nests,with more proliferation of connective tissue.Conclusions:The treatment of matrine combined with cisplatin can significantly improve the anti-tumor effect on U14 rats with cervical cancer,which can be a new option for the treatment for cervical cancer.

Effect of matrine on transforming growth factor β1 and hepatocyte growth factor in rat liver fibrosis model

Objective:To observe the preventive and control effect of matrine on transforming growth factor(TCF- β1) and hepatocyte.growth factor(HCF) of liver fibrosis tissue in rals.Methods:A total of48 SD rats were randomly divided into A,B,C,D groups with 12 in each,group A as the normal control group and groups B.C,D as liver fibrosis models using composite modulus method with carbon tetrachloride(CCL_4).Group B was the model group,group C adopted γ— interferon lavage therapy in the second day of modeling,and group D adopted matrine lavage treatment,at 4 and8 weeks after treatment.Six rats were executed for detection of TGF- β1 and HGF,liver tissue histology and comparison fibrosis degree changes of rat liver tissue between groups.Results:Croups B,C,D showed a more significantly increased TCF- β1 at each time point compared with group A(P<0.05);Group B showed a more significantly increased TGF- β1 than groups C and D at weeks 4 and 8(P<0.05);group D showed a lowest level of TGF-β1,followed by groups C and B.HGF of group B decreased more significantly than A group at weeks 4 and 8(P<0.05);HGF of groups C and D was significantly elevated at 4 and 8 weeks than groups A and B(P<0.05),in which the group D showed the highest level of HGF.According to tissue histologic observation,rat liver tissue structure of group A was clear and normal,tissue structure of group B was destroyed with obvious fibrous tissue hyperplasia and fatty change of hepatic cells;groups C and D showed a slighter liver tissue damage,cell necrosis and connective tissue hyperplasia in collect abbacy than group B with a trend of obvious improvement.Conclusions:Matrine can reduce TGF- β1expression and enhance the activity of HGF,so as to realize the inhibition effect on liver fibrosis in rats.

Anti-HBV effect of liposome-encapsulated matrine in vitro ana in vivo

AIM: To study the anti-HBV effect of liposome-encapsulated matrine (Lip-M) in vitro and in vivo. METHODS: 2.2.15 cell line was cultured in vitro observe the effect of Lip-M and matrine on the secretion of HBsAg and HBeAg. The toxicity of Lip-M and matrine to 2.2.15 cell line was also studied by MTT method. In in vivo study, drug treatment experiment was carried out on the 13th day after ducks were infected with duck hepatitis B virus (DHBV). The ducks were randomly divided into 4 groups with 5-6 ducks in each group. Lip-M and matrine were given to DHBV-infected ducks respectively by gastric perfusion. Four groups were observed: group of Lip-M (20 mg/kg), group of Lip-M (10 mg/kg), group of matrine (20 mg/kg) and group of blank model. The drug was given once daily for 20 d continuously, and normal saline was used as control. The blood was drawn from the posterior tibial vein of all ducks before treatment (T0), after the medication for 5 (T5), 10 (T10), 15 (T15), 20 (T20) d and withdrawl of the drug for 3 d (P3). The serum samples were separated and stored at -70 ℃, DHBV-DNA was detected by the dot-blot hybridization. RESULTS: After addition of Lip-M and matrine to 2.2.15 cell line for eleven d, the median toxic concentration (TC50) of Lip-M and matrine was 7.29 mg/mL and 1.33 mg/mL respectively. The median concentration (IC50) of Lip-M to inhibit HBsAg and HBeAg expression was 0.078 mg/mL and 3.35 mg/mL respectively. The treatment index (TI) value of Lip-M for HBsAg and HBeAg was 93.46 and 2.17 respectively, better than that of matrine. The DHBV-infected duck model treatment test showed that the duck serum DHBV-DNA levels were markedly reduced in the group of Lip-M (20 mg/kg) after treated by gastric perfusion for 10, 15 and 20 d (0.43±0.22 vs 0.95±0.18, t = 4.70, P= 0.001<0.01.0.40±0.12 vs 0.95±0.18, t = 6.34, P= 0.000<0.01. 0.22±0.10 vs 0.95±0.18, t = 8.30, P= 0.000<0.01), compared to the group of matrine (20 mg/kg) (0.43±0.22 vs 0.79±0.19, t = 3.17, P= 0.01<0.05. 0.40±0.12 vs 0.73±0.24, t = 3.21, P= 0.009<0.05. 0.22±0.10 vs0.55±0.32, t = 2.27, P= 0.046<0.05.), and the control (0.43±0.22 vs50.98±0.29, t = 3.68, P = 0.005<0.01. 0.40±0.12 vs 0.97±0.30, t = 4.26, P= 0.002<0.01. 0.22±0.10 vs 0.95±0.27, t = 5.76, P= 0.000<0.01). After the treatment for 20 d and withdrawl of the drug for 3 d, duck serum DHBV-DNA level in the group of Lip-M (10 mg/kg) markedly reduced (0.56±0.26 vs0.95±0.38, t = 5.26, P= 0.003<0.05. 0.55±0.25 vs 0.95±0.38, t = 5.52, P= 0.003<0.05), and the difference was significant as compared with the control (0.56±0.26 vs 0.95±0.27, t = 2.37, P = 0.042<0.05. 0.55±0.25 vs 0.89±0.18, t = 2.55, P= 0.031<0.05), but not significant as compared with the group of matrine (20 mg/kg). After withdrawl of the drug for 3 d, the levels of DHBV-DNA did not relapse in both groups of Lip-M. CONCLUSION: Lip-M can evidently inhibit the replication of hepatitis B virus In vitro and in viva, its anti-HBV effect is better than that of matrine.

Effect of matrine and carvedilol on collagen and MMPs activity of hypertrophy myocardium induced by pressure overload

Objective： To explore the effect and mechanism of matrine （Mt.） on myocardial interstitial fibrosis induced by pressure overload. Methods： Pressure overloaded myocardial hypertrophy was produced by banding of aorta abdominalis in 67 male Sprague-Dawley rats weighing （200±15） g. The rats were assigned into one of the following groups： sham-operation control, operation control, operation group treated with matrine （15 mg/（kg·d）） and treated with carvedilol （Car.） （3.6 mg/（kg·d）） group. The rats were given drugs one day after operation. Five weeks after treatment, the left ventricular weight （LVW） was measured and the volume of myocardial cells was detected with Hematoxylin-Eosin （H-E） stain and Masson stain was used to assess the level of fibrosis of the myocardial matrix. Myocardial metalloproteinase activity was quantified with zymography, and survival rate was calculated. Results： Survival rate significantly decreased （P〈0.05）, LVW/BW （body weight）, MMP-2 （matrix metalloproteinase-2） activity （P〈0.05）, size of cardiomyocytes and interstitial fibrosis obviously increased in the operation group compared with sham control group. Mr. and Car. treatment can significantly increase survival rate （P〈0.05）, decrease LVW/BW （P〈0.05） and MMP-2 activity （P〈0.05）, decrease size of cardiomyocytes and interstitial fibrosis compared with operation group. But there was difference compared with sham group. Conclusion： Matrine was shown to be able to prevent cardiac remodelling of bypertrophy cardium induced by pressure overload including myocardial hypertrophy and fibrosis which may be associated with the decrease in MMP-2 activity of heart.

Matrine promotes neural circuit remodeling to regulate motor function in a mouse model of chronic spinal cord injury

In chronic phase of spinal cord injury, functional recovery is more untreatable compared with early intervention in acute phase of spinal cord injury. In the last decade, several combination therapies successfully improved motor dysfunction in chronic spinal cord injury. However, their effectiveness is not sufficient. We previously found a new effective compound for spinal cord injury, matrine, which induced axonal growth and functional recovery in acute spinal cord injury mice via direct activation of extracellular heat shock protein 90. Although our previous study clarified that matrine was an activator of extracellular heat shock protein 90, the potential of matrine for spinal cord injury in chronic phase has not been sufficiently evaluated. Thus, this study aimed to investigate whether matrine ameliorates chronic spinal cord injury in mice. Once daily intragastric administration of matrine(100 μmol/kg per day) to spinal cord injury mice were starte at 28 days after injury, and continued for 154 days. Continuous mat rine treatment improved hindlimb motor function in chronic spinal cord injury mice. In injured spinal cords of the matrine-treated mice, the density of neurofilament-H-positive axons was increased. Moreover, matrine treatment increased the density of bassoon-positive presynapses in contact with choline acetyltransferase-positive motor neurons in the lumbar spinal cord. These findings suggest that matrine promotes remodeling and reconnection of neural circuits to regulate hindlimb movement. All protocols were approved by the Committee for Animal Care and Use of the Sugitani Campus of the University of Toyama(approval No. A2013 INM-1 and A2016 INM-3) on May 7, 2013 and May 17, 2016, respectively.

Relaxant Effects of Matrine on Aortic Smooth Muscles of Guinea Pigs

Objective To determine whether matrine, a kind of traditional Chinese medicinal alkaloid, can relax the aortic smooth muscles isolated from guinea pigs and to investigate the mechanism of its relaxant effects. Methods Phenylephrine or potassium chloride concentration-dependent relaxation response of aortic smooth muscles to matrine was studied in the precontracted guinea pigs. Results Matrine （1×10^-4 mol/L -3.3×10^-3 mol/L） relaxed the endothelium-denuded aortic rings pre-contracted sub-maximally with phenylephrine, in a concentration-dependent manner, and its pre-incubation （3.3× 10^- 3 mol/L） produced a significant rightward shift in the phenylephrine dose-response curve, but had no effects on the potassium chloride-induced contraction. The anti-contractile effect of matrine was not reduced by the highly selective ATP-dependent K^＋ channel blocker glibenclamide （10.5 mol/L）, either by the non-selective K^＋channel blocker tetraethylammonium （10^-3 mol/L）, or by the β-antagonist propranolol （10^-5 mol/L）. In either ＂normal＂ or ＂Ca^2＋-free＂ bathing medium, the phenylephrine-induced contraction was attenuated by matrine （3.3×10^-3 mol/L）, indicating that the vasorelaxation was due to inhibition of intracellular and extracellular Ca^2＋ mobilization. Conclusion Matrine inhibits phenylephrine-induced contractions by inhibiting activation of α-adrenoceptor and interfering with the release of intracellular Ca^2＋ and the influx of extracellular Ca^2＋.

Synthesis and biological evaluation of nitric oxide-releasing matrine derivatives as anticancer agents

A series of furoxan-based nitric oxide-releasing matrine derivatives(10a-f) were synthesized.The biological evaluation showed that compounds 10a,10b,10e and 10f had stronger cytotoxic activities than 5-fluorouracil against human hepatoma cells(HepG2) in vitro.