1.The Effects of Methontrexate on the Pancreas of Rats: A Histochemical and Ultrastructural Study.
Hong Yul CHOI ; Chung Sook KIM ; Yoo Bock LEE
Yonsei Medical Journal 1969;10(2):117-124
Methonexate is one of the well known anti-cancer chemotherapeutic agents and exerts its action by inhibiting mitoses by inhibition of nucleic acid synthesis. Its effect on actively proliferating normal and pathologic tissues have been well documented. However, little information is available on its effect on tissue which shows no active mitoses but does have a very active metabolic process, such as the pancreas. The present study investigated the histochemical and ultrastructural changes which take place within 24 hours after a single intraperitoneal injection of methotrexate. Using a light microscope for observation, no .specific or constant alteration was noted except for a mild acinar cell dissociation 18 hours after the injection. However, electron microscopic observations showed that several organelles of pancreatic acinar cell revealed ultrastructural changes such as vesiculation and dilation of the cisternae of endoplasmic reticulum, and the appearance of autophagic vacuoles which contained cellular organelles, and showed hyperplasia and dilation of Golgi complexes. The nuclei and zymogen granules were not significantly altered. The changes of endoplasmic reticulum were ,distinctly seen from 1 hour after the injection and were most severe at 6 hours. Autophagic vacuoles appeared at 6 hours and had progressively increased in number and size 18 hours after the injection. Similar changes were also reported in experimental animals which were treated with several cytotoxic agents. According to this study, it is evident that a single administration of methotrexate within short time interval induced a series of ultrastructural alterations in several organelles of the pancreatic acinar cells. It is not clear as yet whether or not this is a specific reaction of cells to methotrexate.
Animals
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Female
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Male
;
Methotrexate/*pharmacology
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Pancreas/cytology/*drug effects
;
Rats
2.Shenghua decoction reversed methotrexate-induced interferon-gamma immunosuppression.
Xia LI ; Xue-zhi CHI ; Li WANG
Chinese Journal of Integrated Traditional and Western Medicine 2011;31(3):363-367
OBJECTIVETo observe the effect of Shenghua Decoction in reversing IFN-gamma inhibition induced by methotrexate (MTX).
METHODSPregnant Balb/C mice were randomly divided into the negative control group, Shenghua Decoction group, the MTX control group, and the MTX combined with Shenghua Decoction group. Pregnant mice in the 4 groups were respectively treated with normal saline (NS), Shenghua Decoction, MTX, and MTX combined with Shenghua Decoction for 7 successive days. Then mice were sacrificed 1 h after the last administration. Living fetus and dead fetus were counted to assess meto-maternal rejection. The percentages of interferon-gamma+(IFN-gamma+) and CD3+CD+IFN-gamma+ cells in spleen lymphocytes of pregnant mice were double stained for cytometric analysis. At the same time IFN-gamma message ribonucleic acid (mRNA) in the spleen mononuclear cells was detected by reverse transcription polymerase chain reaction (RT-PCR).
RESULTSCompared with the negative control group, the number of dead fetus increased significantly after the administration of Shenghua Decoction (P > 0.05). The percentages of IFN-gamma+ and CD3+CD4+IFN-gamma+ in spleen mononuclear cells were elevated by 28.81% and 35.61% respectively (both P < 0.05). IFN-gamma mRNA expression increased (P < 0.05). After the administration of MTX, the number of dead fetus increased significantly. The percentages of IFN-gamma+ and CD3+CD4+IFN-gamma+ in spleen mononuclear cells were reduced by 24.23% and 26.77% respectively compared with the negative control group. Besides, the expression of IFN-gamma mRNA decreased obviously (P < 0.05). However, after the administration of MTX in combination of Shenghua Decoction, the number of dead fetus was further added. Moreover, the percentages of IFN-gamma+ and CD3+CD4+IFN-gamma+ cells obviously increased. The increment was 64.96% and 71.38% respectively when compared with the MTX group, and the expression of IFN-gamma mRNA was obviously up-regulated (P < 0.05).
CONCLUSIONShenghua Decoction showed reversing effect on MTX induced IFN-gamma inhibition by promoting the differentiation of IFN-gamma+Th1 subpopulation and up-regulating IFN-gamma expression, thus promoting the feto-maternal immunological rejection.
Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Immunosuppression ; Interferon-gamma ; immunology ; Methotrexate ; pharmacology ; Mice ; Mice, Inbred BALB C ; Pregnancy
3.Resistance mechanisms to Methotrexate.
Chinese Journal of Pediatrics 2003;41(5):388-390
4.Treatment of Refractory Rheumatoid Arthritis by Huayu Tongbi Recipe Combined Methotrexate.
Xiu-min CHEN ; Run-yue HUANG ; Jing-yao YAN ; Zhi-hong LIU ; Yong-liang CHU ; Qing-chun HUANG
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(11):1326-1330
OBJECTIVETo evaluate the clinical efficacy and safety of Huayu Tongbi Recipe (HTR) combined methotrexate (MTX) in treating refractory rheumatoid arthritis (RRA).
METHODSTotally 167 RRA patients were assigned to the treatment group (73 cases) and the control group (94 cases) according to different therapeutic methods. Patients in the treatment group were treated with HTR combined MTX, while those in the control group were treated with leflunomide (LEF) combined MTX. Clinical signs and symptoms, RF, CRP, ESR, disease activity score 28 (DAS28), and safety indicators were compared between the two groups before treatment, at week 12 and 24 after treatment. The efficacy and safety indices were also evaluated.
RESULTSAt week 12 after treatment the total effective rate was 82.2% (60/73 cases) in the treatment group and 79.8% (75/94 cases) in the control group, showing no statistical difference between the two groups (chi2 = 0.15, P > 0.05). At week 24 after treatment the total effective rate was 78.1% (57/73 cases) in the treatment group and 755% (71/94 cases) in the control group, showing no statistical difference between the two groups (chi2 = 0.15, P > 0.05). There was statistical difference in the total effective rate between week 24 and week 12 in the control group (chi2 = 0.49, P < 0.05). Clinical signs and symptoms, RF, CRP, ESR, and DAS28 were significantly improved in the two groups after 12- and 24-week treatment (P < 0.01). There was no statistical difference in the improvement at week 12 after treatment between the two groups (P > 0.05). There was statistical difference in time of morning stiffness, tender joint numbers, swollen joint numbers, patient global assessment, RF, CRP, and DAS28 at week 24 after treatment between the two groups (P < 0.05). Besides, adverse reactions occurred less in the treatment group than in the control group (P < 0.01).
CONCLUSIONThe efficacy of HTR combined MTX was equivalent to that of LEF (10 mg per day) combined MTX, but with more stable therapeutic effects and less adverse reactions.
Antirheumatic Agents ; pharmacology ; therapeutic use ; Arthralgia ; Arthritis, Rheumatoid ; drug therapy ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Humans ; Isoxazoles ; Methotrexate ; pharmacology ; therapeutic use ; Phytotherapy ; Treatment Outcome
5.Plasma levels of oral methotrexate in children receiving maintenance chemotherapy for acute lymphocytic leukemia.
Soo Kwan LEE ; Kih Yeon SONG ; Young Hee HWANG ; Young Hwan LEE ; Jeong Ok HAH ; Chun Dong KIM
Journal of the Korean Pediatric Society 1993;36(7):936-943
This study was conducted to investigate plasma levels of oral methotrexate in rabbits and children receiving maintenance chemotherapy for acute lymphocytic leukemia. Eight New Zealand white rabbits, weighing 2kg in body weight, were divided into 3 groups and 5mg of methotrexate from 3 different manufactorying company was administered to the each group rabbits via nasogastric tube. Time to peak concentration ranged from 30 minutes to 3 hours (mean 1.2+/-0.9 hour)and the peak plasma concentration ranged from 0.08 micro M to 0.21micron M(mean 0.14+/-0.05 micronM)and area under the plasma concentration-time curve ranged from 0.6micron M.hr to 1.66micron M,hr (mean 1.06+/-0.36micronM,hr). There were no statistically significant difference in AUC of methotrexate in 3 groups, but interindividual variability in plasma levels of methotrexate was noted. Twelve patients with ALL who were receiving maintenance chemotherapy at pediatric department of Yeungnam University Hospital from August, 1988 to August, 1991 were studied. Plasma levels of oral methotrexale were monitored following an oral dose of 3.3 mg~10mg/m2 which was modified from recommended dose of 10 mg/m2 due to hepatotoxicity or myelosuppression. Time to peak concentration ranged from 30 minutes to 2 hours(mean 1.2+/-0.4 hour) and the peak plasma concentration ranged from 0.34 micron M to 0.8 micron M (mean 0.58+/-0.18micron M). The area under the plasma concentration-time curve ranged from 1.25micron M,hr to 3.79 micronM,hr (mean 2.71+/-0.84microM,hr)while standard area under the plasma concentration-time curve ranged from 0.13micronM, hr/mg/m2 to 0.54micronM, hr/mg/m2 (mean0.4+/-0.15micronM hr/mg/m2).Interindividual variability in plasma levels following an oral dose of methotrexate was noted. Peak plasma concentrations of study patients were all less than 1 micronM which is necessary for antileukemic effect of methotrexate in vitro. It seems to be necessary to increase the dose of methotrexate for all study patients, however optimal dose increment of methotrexate avoiding hepatotoxicity and myelosuppression need to be investigated further and measurement of plasma level of methotrexate is recommended when dose modification of methotrexate is made.
Area Under Curve
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Body Weight
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Child*
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Humans
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Maintenance Chemotherapy*
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Methotrexate*
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Pharmacology
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Plasma*
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Precursor Cell Lymphoblastic Leukemia-Lymphoma*
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Rabbits
6.Effects of sinomenine and methotrexate on fibroblast-like synoviocytes in rheumatoid arthritis.
Yue SUN ; Cong-zhu DING ; Yao YAO
Chinese Journal of Integrated Traditional and Western Medicine 2012;32(8):1107-1111
OBJECTIVETo investigate the effects of sinomenine (SIN) and methotrexate (MTX) on the proliferation and apoptosis of in vitro cultured fibroblast-like synoviocytes (FLS) in rheumatoid arthritis (RA) patients, as well as the expression of osteoclast differentiation factor in FLS.
METHODSFLS were isolated from the synovium of RA patients and cultured in vitro. FLS were incubated with different concentrations of SIN and MTX respectively or combined: 0.001, 0.010, 0.100, 1.000 mg/mL SIN; 0.001, 0.010, 0.100, 1.000 mg/mL MTX; 0.001 mg/mL SIN + 0.001 mg/mL MTX, 0.010 mg/mL SIN + 0.010 mg/mL MTX, 0.100 mg/mL SIN + 0.100 mg/mL MTX, 1.000 mg/mL SIN + 1.000 mg/mL MTX, namely SIN1, 2, 3, 4 groups; MTX1, 2, 3, 4 groups and the combination 1, 2, 3, 4 groups. The medium without drugs was used as a control group. There was a total of 13 groups, each group with 3 complex holes. MTT was applied to detect the growth of FLS. The flow cytometry was applied to detect the apoptosis of FLS. The expressions of FLS receptor activator of nuclear factor kappa B ligand (RANKL) mRNA and osteoprotegerin (OPG) mRNA were observed by semi-quantitative RT-PCR.
RESULTSCompared with the control group, RA FLS proliferation OD values of all the drug groups were lower (P < 0.05). The RA FLS apoptosis OD value of the combination 3 group increased, the OPG mRNA expression increased, the expression of RANKL mRNA decreased with statistical difference (P < 0.05). The RA proliferation OD values of the SIN3 group and the MTX3 group increased when compared with the combination 3 group (P < 0.05).
CONCLUSIONSSIN and MTX had synergistic effects in inhibiting FLS. This might be one of the mechanisms for inhibiting RA bone damage.
Apoptosis ; drug effects ; Arthritis, Rheumatoid ; pathology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Fibroblasts ; drug effects ; Humans ; Methotrexate ; pharmacology ; Morphinans ; pharmacology ; Synovial Membrane ; cytology ; drug effects
7.Synthesis of methotrexate-poly (ethylene glycol) conjugate and their anti-tumor activity in vitro.
Jian-Feng ZHANG ; Dong-Zhi WEI ; Xiong ZHOU ; Feng JIANG
Acta Pharmaceutica Sinica 2007;42(6):607-610
To improve the physical property and bioactivity of methotrexate, this paper investigated the new formation of conjugate methotrexate-poly (ethylene glycol) and in vitro anti-tumor activity of the synthesized conjugate. The conjugate of methotrexate-poly (ethylene glycol), which was verified by the spectroscopy analysis of UV, IR and 13C NMR, was synthesized by chemical catalysis and micro-wave irritation. The determination for the conjugate of solubility in water and distribution coefficient in octanol-water system of the conjugate was done to examine its deliquescence property. The solubility in water and the distribution coefficient of the conjugate was greatly improved, which was increased by 128 folds and 5 folds, respectively. The in vitro anti-tumor activity of the conjugate was tested by mouse L(1210) leukaemia cells, and the synthesized conjugate showed the same anti-tumor activity as the original methotrexate. Compared to the reported literature, the modification of methotrexate by poly (ethylene glycol) is more rapid and convenient.
Animals
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Antineoplastic Agents
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chemical synthesis
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Leukemia L1210
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drug therapy
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Methotrexate
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chemical synthesis
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chemistry
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pharmacology
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Mice
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Polyethylene Glycols
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chemical synthesis
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chemistry
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pharmacology
;
Solubility
8.Effects of zhengqing fengtongning tablet and methotrexate on the serum OPG/RANKL and IL-17 of collagen-induced arthritis rats.
Cong-Zhu DING ; Yao YAO ; Yun FANG ; Ling-Yun SUN ; Yue WANG
Chinese Journal of Integrated Traditional and Western Medicine 2013;33(2):256-260
OBJECTIVETo study the effects of Zhengqing Fengtongning Tablet (ZFT) and methotrexate (MTX) on the expression of osteoprotegerin (OPG), receptor activator of nuclear factor-kappaB ligand (RANKL), and interleukin 17 (IL-17) in collagen-induced arthritis (CIA) rats, thus addressing their bone protection.
METHODSThe CIA rat model was established by intradermally injecting type II collagen emulsion from the rats' back and tail. Totally 28 successfully modeled rats [with the arthritis index (AI) more than 2] were randomly divided into the model group, the Chinese medicine (CM) treatment group, the MTX group, and the ZFT + MTX treatment group, 7 rats in each group. Another 7 rats were recruited as the normal control group. Rats were administered from the 7th day of modeling. Rats in the MTX group were treated with MTX at 3.8 mg/kg once a week. Those in the CM group were treated with ZFT at the daily dose of 130 mg/kg, once a day. Those in the ZFT + MTX treatment group were treated with both MTX (at 3.8 mg/kg once a week) and ZFT (at the daily dose of 130 mg/kg, once a day). Those in the model group and the normal control group were administered with normal saline of the equal volume by gastrogavage. All the intervention lasted for 26 days. The destruction of joints in the four limbs were observed using X-ray. The AI was recorded. The expression levels of serum OPG, RANKL, and IL-17 were detected at the end of the experiment.
RESULTSDuring the whole process, all rats except those in the model group were in a good condition. On the 21st day of modeling the AI of all rats reached the peak, but it decreased after treatment. Compared with the model group, the AI decreased in the CM treatment group, the MTX group, and the ZFT + MTX treatment group with statistical difference (P < 0.05). Compared with the model group, the OPG increased and RANKL decreased in the MTX group; the OPG and OPG/RANKL increased in the CM treatment group; the OPG, RANKL, and OPG/RANKL increased, and IL-17 decreased in the ZFT + MTX treatment group, all showing statistical difference (P < 0.05). Compared with the MTX and the ZFT + MTX treatment group, OPG/RANKL increased and IL-17 decreased in the ZFT + MTX treatment group (both P < 0.05).
CONCLUSIONZFT + MTX could synergistically elevate peripheral OPG/RANKL and down-regulate IL-17 in CIA model rats.
Animals ; Arthritis, Experimental ; blood ; chemically induced ; drug therapy ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Interleukin-17 ; blood ; Methotrexate ; pharmacology ; therapeutic use ; Osteoprotegerin ; blood ; RANK Ligand ; blood ; Rats
9.Studies on concentrations and interactions of drugs in patients with administration of high-dose of cytosine arabinoside and methotrexate.
Yan-ning QU ; Bin JIANG ; Yu-hang WANG
Chinese Journal of Hematology 2012;33(12):1049-1051
Adolescent
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Adult
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Cytarabine
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administration & dosage
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blood
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pharmacology
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Drug Interactions
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Female
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Humans
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Instillation, Drug
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Leukemia
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blood
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drug therapy
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Male
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Methotrexate
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administration & dosage
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blood
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pharmacology
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Middle Aged
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Young Adult
10.Purification and activity evaluation of methionine synthase.
Ying GUO ; Chao LI ; Zhi-Li ZHANG ; Chao TIAN ; Xiao-Wei WANG ; Jun-Yi LIU
Acta Pharmaceutica Sinica 2012;47(11):1463-1469
Methionine synthase (MS, EC2.1.1.13), a key enzyme in the folate metabolism area catalyzing methyl transfer from N5-methyltetrahydrofolate to homocysteine to give tetrahydrofolate and methionine, takes a core position in folate cycle, one-carbon-unit transfer and sculpture amino acid pathways. Cobalamin-dependent methionine synthase was purified from rat liver. The enzyme was purified 609-fold to near homogeneity by batch chromatography on DE-52, anion-exchange chromatography on Q Sepharose Fast Flow and CHT-I hydroxyapatite column and was identified by SDS-PAGE and Western blotting. The enzyme activity was determined by spectrophotometric assay. In addition, the influencing factor and optimal reaction condition were performed. The steady state kinetic of rat liver methionine synthase was similar to that of other mammalian cobalamin-dependent methionine synthase which employed a Ping-Pong mechanism. The result indicated that cobalamin-dependent methionine synthase purified from rat liver is suitable for screening and studying methionine synthase specific inhibitors.
5-Methyltetrahydrofolate-Homocysteine S-Methyltransferase
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isolation & purification
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metabolism
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Animals
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Electrophoresis, Polyacrylamide Gel
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Folic Acid Antagonists
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pharmacology
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Liver
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chemistry
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Male
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Methotrexate
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pharmacology
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Quinazolines
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pharmacology
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Rats
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Rats, Wistar
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Tetrahydrofolates
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metabolism
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Thiophenes
;
pharmacology