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Objective To investigate the epidemiological characteristics of SARS-CoV-2 pneumonia in kidney transplant recipients and analyze the risk and protective factors of severe/critical infection with SARS-CoV-2. Methods Clinical data of 468 kidney transplant recipients infected with SARS-CoV-2 were retrospectively analyzed. According to the severity of infection, they were divided into mild SARS-CoV-2 infection recipients (n=439) and SARS-CoV-2 pneumonia group (n=29). Among the 439 mild SARS-CoV-2 infection recipients, 87 recipients who were randomly matched with their counterparts in the SARS-CoV-2 pneumonia group according to sex, age and transplantation time at a ratio of 3∶1 were allocated into the mild SARS-CoV-2 infection group. Twenty-nine recipients in the SARS-CoV-2 pneumonia group were divided into the moderate SARS-CoV-2 pneumonia group (n=21) and severe/critical SARS-CoV-2 pneumonia group (n=8). Baseline data of all recipients were collected. The risk and protective factors of SARS-CoV-2 infection in kidney transplant recipients were identified. Results The proportion of recipients complicated with 2-3 types of complications in the SARS-CoV-2 pneumonia group was higher than that in the mild SARS-CoV-2 infection group, and the proportion of recipients treated with tacrolimus(Tac)+mizoribine+glucocorticoid immunosuppression regimen in the SARS-CoV-2 pneumonia group was lower than that in the mild SARS-CoV-2 infection group, and significant differences were observed (both P<0.05). In 29 kidney transplant recipients with SARS-CoV-2 pneumonia in the SARS-CoV-2 pneumonia group, white blood cells, the absolute values of lymphocytes, eosinophils, total T cells, CD4+T cells and CD8+T cells, and serum uric acid levels were significantly lower, whereas ferritin levels were significantly higher than the values prior to SARS-CoV-2 pneumonia, and significant differences were observed (all P<0.05). Compared with the moderate SARS-CoV-2 pneumonia group, the proportion of recipients with hypoxemia was higher, the proportion of recipients treated with Tac/ciclosporin (CsA)+mycophenolate mofetil+glucocorticoid immunosuppression regimen was higher, and the proportion of recipients administered with 2-3 doses of SARS-CoV-2 vaccine was lower in the severe/critical SARS-CoV-2 pneumonia group, and significant differences were observed (all P<0.05). Conclusions More complications and immunosuppression regimen containing mycophenolate mofetil are the risk factor for SARS-CoV-2 infection in kidney transplant recipients. Vaccination with SARS-CoV-2 vaccine and immunosuppression regimen containing mizoribine are probably the protective factors for lowering the risk of SARS-CoV-2 infection. The levels of inflammatory cytokines are associated with the severity of SARS-CoV-2 pneumonia.
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Objective: Immunosuppressive therapy for interstitial lung disease (ILD) is often necessary, but the standard regimen for antisynthetase-associated ILD has not been established.Patient: An 80-year-old man was hospitalized for severely progressive dyspnea. Bilateral interstitial shadows occurred 1 month before the event. Serological findings showed that he had antisynthetase-associated ILD, as identified by strong positivity for anti-aminoacyl-transfer RNA synthetase (ARS) antibody, despite no evidence of myositis. He was treated transiently with noninvasive positive pressure ventilation and steroid-pulse therapy followed by 60 mg/day of oral prednisolone. However, his diabetes mellitus was aggravated by corticosteroid therapy; thus, a combination of low-dose steroid and mizoribine (MZB), which has a low risk of aggravating glucose intolerance, was used.Results: The patient’s clinical symptoms and daily life activities have been well persevered as an outpatient and well maintained with 200 mg of MZB and 10 mg of prednisolone for several months without obvious clinical recurrence and without any remarkable steroid- and MZB-related side effects.Conclusion: The use of MZB appeared to suppress the pathophysiology of anti-ARS antibody-associated ILD.
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Objective To assess the efficacy and safety of mizoribine (MZR) in initial immunosuppression in living-related renal transplant recipients.Methods From October 2015 to October 2017,twenty-two patients undergoing initial living-related renal transplantation received MZR (3-4 mg/kg/d) plus tacrolimus and corticosteroid.During a follow-up period of 12 months,patient/graft survival,incidence of acute rejection and adverse events were observed.Results There was no onset of graft loss and death and acute rejection rate was 22.7％.Renal allograft function remained stable.The incidence rate of cytomegaloviral infection was 4.5％ and no CMV disease occurred.The incidence of BKV viruria was 36.4％ and the infection rate was 18.2％.Digestive symptoms occurred (n =3,13.6％).The major side effect of hyperuricemia could be controlled without reduction or withdrawal of MZR.Conclusions Excellent graft survival can be achieved when using MZR as initial immunosuppression in living-donor renal transplant recipients,yet the incidence of acute rejection remains high.Further study is required for determining the effect of MZR in the prevention of BK viral infection during renal transplantation.
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Mizoribine(MZR), as an orally prescribed immunosuppressive agent, has been applied in the prevention of rejection after kidney transplantation.MZR requires individual dosing due to the variation of bioavailability.However, therapeutic drug monitoring (TDM) of MZR is not well developed in China, as compared to other clinically used immunosuppressive agents.To our knowledge, this is the first TDM review of MZR.Pharmacokinetic characteristic, concentration determination methods and sample selection of MZR were summarized, also the rational therapeutic window was proposed.Furthermore, gene polymorphism and population pharmacokinetics of MZR were estimated.This review will provide reference for TDM-based individual dosing of MZR in renal transplant recipients.
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Objective To analyze the incidence of BK virus (BKV) infection after kidney transplantation in our center and to evaluate the efficacy and safety of conversion treatment with Mizoribine (MZR) on BKV infection after kidney transplantation.Methods The information of recipients who received BK virus screening in hospital or outpatient during 2015-02 to 2016-12 in our center was retrospectively analyzed.The recipients positive for BKV were divided into experiment group (given conversion treatment with MZR) and control group (not given MZR conversion) according to the inclusion criteria.The negative rate of BKV,AR,hyperuricemia and the function of renal allograft during the conversion treatment with MZR were observed.Results 182 recipients accepted BKV screening during 2015-02 to 2016-12 and 68 cases were positive.The positive rate of BKV was 38.5 ％.The positive rate of peripheral blood specimens and midstream urine specimens was 7.1％ and 36.8％ respectively.Twelve recipients were positive for BKV in both peripheral blood specimens and midstream urine specimens.There were 27 recipients in experiment group and 36 cases in control group.Fourteen recipients positive for BKV became negative after MZR conversion in experiment group and the negative rate was up to 51.9％.The mean time of negative rate was 3.2 ± 2.7 (1-10) months after MZR conversion.During the conversion treatment with MZR,AR occurred in 1 case and was reversed by the impact therapy with Thymoglobulin in experiment group.The value of serum uric acid was maintained stable before and after MZR conversion under the action of uric-acidlowering drug.The renal function was kept stable in both experiment group and control group after renal transplantation.No deaths and renal allograft failure cases occurred in both groups during the research period.The 2-year survival rate for patients and kidneys was both 100％.Conclusion The incidence of BKV infection after kidney transplantation was high and the treatment scheme of MZR conversion was safe and effective.
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Objective To evaluate the effect of conversion from mycophenolic acid (MPA) to mizoribine (MZR) in renal transplant recipients with gastrointestinal tract (GI) symptoms.Methods A total of 355 renal transplant recipients with GI symptoms caused by MPA administration were enrolled from April 2015 to March 2017 in 25 different renal transplant centers in China.The symptomatic improvement of GI before (baseline) and after conversion to MZR (1,2,4 weeks) was assessed by each item of GI symptoms indication.In addition,the efficacy and safety of the conversion therapy during 12 months were determined.Results Patients showed improvement in GI symptoms including diarrhea,abdominal pain,abdominal distention and stomachache after conversion to MZR 1,2,4 weeks (P＜0.05).In patients with different severity of diarrhea,conversion to MZR therapy significantly improved diarrhea (P＜0.05).During 12 months,no patient experienced clinical immune rejection.We did not observe any infections,leucopenia and other serious side effects.Conclusion MZR could markedly improve GI symptoms caused by MPA administration in renal transplant recipients.
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BACKGROUND: Mizoribine (MZR) is an immunosuppressive drug used in Japan for treating patients with lupus nephritis and nephrotic syndrome and has been also reportedly effective in patients with immunoglobulin A (IgA) nephropathy. However, to date, few randomized control studies of MZR are performed in patients with IgA nephropathy. Therefore, this prospective, open-label, randomized, controlled trial aimed to investigate the efficacy and safety of adding MZR to standard treatment in these patients, and was conducted between April 1, 2009, and March 31, 2016, as a multicenter study. METHODS: Patients were randomly assigned (1:1) to receiving standard treatment plus MZR (MZR group) or standard treatment (control group). MZR was administered orally at a dose of 150 mg once daily for 12 months. RESULTS: Primary outcomes were the percentage reduction in urinary protein excretion from baseline and the rate of patients with hematuria disappearance 36 months after study initiation. Secondary outcomes were the rate of patients with proteinuria disappearance, clinical remission rate, absolute changes in estimated glomerular filtration rate from baseline, and the change in daily dose of prednisolone. Forty-two patients were randomly assigned to MZR (n = 21) and control groups (n = 21). Nine patients in MZR group and 15 patients in the control group completed the study. No significant differences were observed between the two groups with respect to primary and secondary outcomes. CONCLUSION: The addition of MZR to standard treatment has no beneficial effect on reducing urinary protein excretion and hematuria when treating patients with IgA nephropathy.
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Humans , Glomerular Filtration Rate , Glomerulonephritis, IGA , Hematuria , Immunoglobulin A , Immunoglobulins , Japan , Lupus Nephritis , Nephrotic Syndrome , Prednisolone , Prospective Studies , ProteinuriaABSTRACT
Objective To observe the effects of mizoribine(MZR)on renal tubular epithelial?mesenchymal transition(EMT)of mice which have been performed unilateral ureteral obstruction(UUO),and study the mechanism of its anti?fibrosis of renal interstitial. Methods A total of 24 CD1 mice were randomly divided into sham group,UUO model group and MZR treatment group,with 8 mice in each group. The day before op?eration,mice of MZR treatment group had been given MZR 10 mg/kg/d lavage,those of sham group and UUO model group had been given equal saline lavage. Fourteen days after the operation ,blood was collected and serum creatinine and blood urea nitrogen were measured;the obstruction kidneys were harvested for section,HE staining and Masson staining were employed to observe the changes of kidney pathological;the expression ofα?SMA and E?Cad in kidney with detected by immunohistochemical and Western blot method. Results Compared with sham group,serum creatinine and blood urea nitrogen of mice in UUO model group and MZR treatment group were significantly elevated ,kidney pathological chang?es and the expression ofα?SMA in renal tissue were increased and that of E?Cad was reduced ,the differences were all statistically significant(P<0.05);compared with UUO model group,mice in MZR treatment group had different degree of improvements in serum creatinine,blood urea ni?trogen and kidney pathological changes ,the expression ofα?SMA in renal tissue was inhibited and that of E?Cad was increases ,and the differences were statistically significant(P<0.05). Conclusion MZR may inhibit the development of renal tubular EMT in UUO mice ,thereby reduce the level of renal tubule interstitial fibrosis and improve renal function.
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After invasion of red blood cells, malaria matures within the cell by degrading hemoglobin avidly. For enormous protein breakdown in trophozoite stage, many efficient and ordered proteolysis networks have been postulated and exploited. In this study, a potential interaction of a 60-kDa Plasmodium falciparum (Pf)-heat shock protein (Hsp60) and Pf-calpain, a cysteine protease, was explored. Pf-infected RBC was isolated and the endogenous Pf-Hsp60 and Pf-calpain were determined by western blot analysis and similar antigenicity of GroEL and Pf-Hsp60 was determined with anti-Pf-Hsp60. Potential interaction of Pf-calpain and Pf-Hsp60 was determined by immunoprecipitation and immunofluorescence assay. Mizoribine, a well-known inhibitor of Hsp60, attenuated both Pf-calpain enzyme activity as well as P. falciparum growth. The presented data suggest that the Pf-Hsp60 may function on Pf-calpain in a part of networks during malaria growth.
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Humans , Amino Acid Sequence , Calpain/genetics , Chaperonin 60/chemistry , Erythrocytes/parasitology , Malaria, Falciparum/parasitology , Molecular Sequence Data , Plasmodium falciparum/chemistry , Protein Binding , Protozoan Proteins/chemistry , Sequence AlignmentABSTRACT
Background: Mizoribine (MZR) is an antimetabolite that inhibits inosinemonophosphate dehydrogenase and has been used for preventing rejection in renal transplantation. However, the effect of calcineurin inhibitors (CNIs) on the pharmacokinetics of MZR has not been shown. This study was performed to show the influence of CNIs (tacrolimus [Tac] or cyclosporine [CyA]) on the serum concentration of MZR. Methods: Thirty-four living-donor renal transplant recipients administered a fourdrug immunosuppressive therapy regimen (steroid, CNIs, basiliximab and MZR 6 mg/kg/day) were investigated. 20 recipients were treated with Tac and 14 were with CyA. Serum concentrations of MZR were obtained retrospectively at 464 points and at 243 points for each. Population pharmacokinetic (PPK) analysis was used to make pharmacokinetic models of serum MZR. After statistically evaluating the correlation of the pharmacokinetic models with the actual data, areas under the curves (AUCs) of each CNI were also estimated in these models and statistically evaluated. Results: The mean values of the PPK parameters (absorption lag time, absorption rate constant [Ka], apparent volume of distribution [V/F] and oral clearance of MZR [CLMZR/F]) were 0.600 hr and 0.643 hr, 1.14/hr and 0.911/hr, 0.732×body weight (WT) (L) and 0.784×WT (L), and 1.64×creatinine clearance (CLcr) (L/hr) and 1.81×CLcr (L/hr), respectively. Moreover, the serum concentrations of MZR at all-time points were estimated with these parameters. The correlation coefficients between the individual actual and estimated serum concentrations of MZR in the Tac group and the CyA group were 0.988 and 0.992, respectively. The average value of the AUCs of MZR corrected by the CLcr in the Tac group, and the CyA group were 0.61±0.21 and 0.55±0.19 (average value±standard deviation) for each (p=0.19). Conclusion: These findings suggest the pharmacokinetics of MZR were welldescribed by 1-compartment model with first-order absorption. Moreover, concomitant use of CNIs, e.g., Tac and CyA, may have no significant influence on the pharmacokinetics of MZR.
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Objective Mizoribine ( MZR) is a new immunosuppressant , however , little domestic research has been done on MZR for treatment of nephrotic syndrome in children .This study was to investigate curative effect and adverse reaction of MZR in the treatment of children with frequently relapsing nephrotic syndrome , using prospective controlled trials . Methods A total of 59 pa-tients with frequency relapsing nephrotic syndrome were randomly divided into two groups .29 patients of treatment group were treated with MZR +glucocorticoid , while 30 patients of control group were given Tripterygium wilfordii ( TW)+glucocorticoid treatment , and the course of treatment lasted for 12 months.24-hour urine protein, urinary N-acetyl β-glucosidase (NAG), serum albumin, serum cholesterol, serum creatinine, recurrence frequency, and average prednisone dosage were observed . Results At the end of treat-ment, Serum albumin in treatment group was higher than that in control group [(40.95 ±6.12)g/L vs (30.25 ±9.02)g/L], and Se-rum cholesterol ([5.45 ±0.82]mmol/L vs [7.53 ±2.74]mmol/L), urinary protein ([0.89 ±0.52]g/24 h vs [1.63 ±2.02]g/24 h), urinary NAG enzyme ([21.43 ±14.16]U/g· Cr vs [41.67 ±12.35]U/g· Cr) levels were lower compared with control group . There was significant difference between the two groups .In terms of mean recurrence times , no significant difference was found at 6th months of follow-up between the two groups, however, treatment group had lower recurrence rate than control group at 3rd month, 9th month, 12th month of follow-up, which was of significant difference .The average amounts of hormone of treatment group were lower than those of control group ([0.56 ±0.16] mg/kg· d vs [0.72 ± 0.34]mg/kg· d)、([0.64 ±0.35]mg/kg· d vs [0.67 ±0.52]mg/kg· d)、([0.53 ±0.41] mg/kg· d vs [0.83 ±0.37] mg/kg· d)、([0.34 ±0.15] mg/kg· d vs [0.54 ±0.26] mg/kg· d) at 3rd month, 6th month, 9th month, 12th month of follow-up, which was of significant difference . Conclusion Compared to Tripterygium wil-fordii combined with hormone therapy , MZR combined with prednisone therapy in children with recurrent NS frequency can reduce the relapse rate and dosage of corticosteroid to improve the clinical remission rate .
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Objective To observe urinary protein,serum albumin,cholesterol and recurrence of children with frequency recurrence nephrotic syndrome(NS) after using Mizorbine (MZR).Methods Eighteen cases of recurrent frequency NS were given enough hormone and MZR.Urinary protein,serum albumin,cholesterol,urinary N-acetyl-β-D-glucosaminidase(NAG) and recurrence were observed.The difference of recurrence between the children treated with steroid and the children treated with steroid and MZR more than 6 months were compared.Results In the combined treatment with both sufficient hormone and MZR,out of the 18 cases,there were one lost case and one invalid case.For the remaining 16 cases,edema subsided when they were reexamined in the first month,and the urine protein changed to negative within about 3 to 10 days (the average time was 6.33 days).There were 10 cases who were reexamined after 6 months,in which for 8 cases the serum albumin returned to normal in the first month (≥35 g/L),1case returned to normal in 3 month and 1 case in 6 month.The level of blood cholesterol returned to normal for 7 cases in the first month (＜ 5.7 mmol/L),3 cases returned to normal in 3 months.In the reexamining of urine protein in 24hours in the first month,7 cases returned to normal (＜0.4 g),2 cases reduced and there was no change in 1 case.Relapse occurred in 2 cases,in one of which the level of urine protein returned to normal if changing the MZR to alternate-day implosive therapy.After treatment,partial tubular functions restored and in the first month,the level of NAG returned to normal in 7 cases,decreased in 9 cases and increased in 1 case.In the experiment group,no child was observed vomiting,to have diarrhea and other gastrointestinal symptoms.The blood pressure,breathing and heart rate were within the normal range before and after the treatment.The increase of urea nitrogen and creatinine was observed only in 1 case.Conclusions MZR with enough hormone treating frequency recurrence NS can promote urine protein negative,and make serum albumin,cholesterol,urinary NAG decrease,and reduce the recurrence.When MZR made regular doses of the application can not effectively reduce the recurrence frequency,and the pulse treatment may be a kind of effective methods.
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Mizoribine,an immunosuppressive agent,plays a role in cell synthesis process,selectively inhibits lymphocyte proliferation,and suppresses immune function.It has obtained certain curative effect in resistance the rejection after kidney transplantation,slowing down lupus nephritis pathological progress,reducing the proteinuria and the risk of glomerular sclerosis on IgA nephropathy,decreasing the recurrence frequency and proteinuria on steroid-dependent nephrotic syndrome and reducing proteinuria level on Henoch-Schoenlein purpura nephritis.It's safer,fewer side effects,but for lacking evidences,large sample still be needed.
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Objective To observe the efficacy and safety of different doses of mizoribine to prevent rejection after renal transplantation. Methods Sorted by time of operation and odevity, 206 primary kidney transplant recipients were divided into 3 groups, including MMF group, MZR Ⅰ group and MZR Ⅱ group. All recipients in 3 groups were administrated CsA and Pred, combined with mycophenolate mofitile (MMF) in MMF group and mizoribine (MMF) in MZR Ⅰ and Ⅱ groups.The dosage of MMF was 1. 0 g/day, while dosage of MZR in MZR Ⅰ and Ⅱ groups was 100 and 200 mg/day, respectively. There was no difference in usage of cyclosporine (CsA) and prednisone (Pred) among 3 groups. 100, 60 and 30 recipients were followed up in MMF, MZR Ⅰ and MZR Ⅱ groups respectively in 5 years. During the follow-up period of 5 years, the incidence of acute rejection, patient/graft survival and adverse effects associated with drugs in three groups were observed. Results The patient/graft survival was 88. 3 % (53/60), 85 % (51/60) in MZR Ⅰ group, 90 % (27/30),86.7 % (26/30) in MZR Ⅱ group, and 88% (88/100), 86% (86/100) in MMF group, respectively (P>0. 05). There was no significant difference in incidence of acute rejection among MZR Ⅰ (10 %, 6/60), MZR Ⅱ (6. 7 %, 2/30) and MMF groups (9 %, 9/100). The incidence of severe pulmonary infection in MZR Ⅰ group was 3. 3 % (2/60), and 10 % (3/30) in MZR Ⅱ , and the former was lower than MMF group (15 %, 15/100) significantly. There was significant difference in mortality of severe pulmonary infection between MZR Ⅰ group (0, 0/2) and MMT group (73. 3 %, 11/15). The rate of ACR in MZR Ⅱ group (10 %, 3/30) was lower significantly than MMF group (30 %, 30/100) and MZR Ⅰ group (31.7 %, 19/60). There was significant difference in the incidence of hyperuricacidemia between two MZR groups (30 %, 56. 7 %) and MMF group (10 %)(P<0. 05), while the incidence of diarrhea and myelosuppression was lower significantly in MZR Ⅰ group than in MMF group. Conclusion MZR can prevent acute rejection after kidney transplantation effectively and safely. Immunosuppressive therapy including mizoribine is the best choice especially for high risk group because of susceptibility to infection and those who suffer from tenacious diarrhea owing to the side effect.
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Objective To investigate the efficacy and safety of conversion therapy to mizoribine (MZR) for renal transplant patients who suffered MMF or Aza adverse reaction. Methods In 56 patients with adverse reactions at different time points after renal transplantation, there were 23 cases of pulmonary infection, 14 cases of bone marrow depression, 6 cases of hepatic functional lesion and 13 cases of diarrhea. The immunosuppressive protocols of these patients were changed to CNI + MZR + Pre when the adverse reaction occurred. During the follow-up period (11 to 53 months), the effect and adverse events of conversion treatment were observed. Results After conversion treatment, 1 of 23 patients with pulmonary infection was re-infected after 26 months and finally died of heart and lung function failure. In 14 patients with bone marrow depression, blood test returned to normal in 13cases. Six patients with hepatic functional lesion were administered hepatoprotection treatment and their liver function was restored without recurrence of impaired liver function. All 13 patients with diarrhea were relieved without recurrence. The serum creatinine was 123 ± 21.3 μmol/L and 119±18. 2 μmol/L before and after the conversion therapy respectively (P＞0. 05). During the follow-up period, all patients' graft function was good. The incidence of rejection was 1.7 % (1 case). Nine patients (16. 1 %) had a higher level of uric acid after conversion. One patient had finger and toe joint pain. The symptoms were relieved after symptomatic treatment. Conclusion There were high security and good effect of conversion therapy to MZR due to MMF or Aza adverse reaction. Besides, MZR conversion therapy for renal transplantation patients provided a new option for individual immunosuppression.
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Hand-foot syndrome is also known as palmar-plantar erythrodysesthesia, and it is a side effect that mostly occurs with chemotherapy or biologic therapy. It is characterized by a painful erythema and edema, which may be followed by dry or moist desquamation of the palms and soles. Mizoribine (MZR) is an imidazole nucleoside that blocks purine biosynthesis. It was found to inhibit both humoral and cellular immunity by selectively inhibiting proliferation of lymphocytes. MZR has been used for preventing renal transplant rejection and treating rheumatoid arthritis, nephritic syndrome and lupus nephritis. The principal adverse reactions are leukopenia, abnormal hepatic function and hyperuricemia. The cutaneous side effects are skin rash, prurigo and epilation, with an incidence of about 1.32%, 0.77% and 0.55%, respectively. To the best of our knowledge, no case of hand-foot syndrome associated with MZR has ever been published. Herein, we report that MZR, which has been widely used, may induce drug-induced hand-foot syndrome.
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Arthritis, Rheumatoid , Biological Therapy , Edema , Erythema , Exanthema , Graft Rejection , Hair Removal , Hand-Foot Syndrome , Hyperuricemia , Imidazoles , Immunity, Cellular , Incidence , Leukopenia , Lupus Nephritis , Lymphocytes , Prurigo , Purines , RibonucleosidesABSTRACT
BACKGROUND: This retrospective study was performed to investigate the clinical effects of mizoribine with using methotrexate (MTX) or mizoribine alone on those patients with rheumatoid arthritis (RA) who showed an ineffective response or intolerance to the MTX. METHODS: The patients were divided into two groups: (1) combination therapy of mizoribine with MTX and (2) mizoribine alone. All the patients took 100 mg mizoribine daily for at least 16 weeks. Before and after administration of mizoribine for 16 weeks, we assessed the clinical variables such as the visual analogue pain scale (VAS), the tender joint counts (TJC), and the swollen joint counts (SJC). At each time, the laboratory parameters including the ESR, CRP, complete blood count (CBC), liver enzymes and creatinine were also measured. Disease activity scores (by the DAS28) and the adverse effects were determined at baseline and after 16 weeks. RESULTS: Fifty patients were recruited in this study (mizoribine plus MTX group: n=35, mizoribine group: n=15). There were no significant differences in the initial laboratory values between the two treatment groups. After treatment for 16 weeks, the DAS28 was decreased significantly in the mizoribine plus MTX group (4.7+/-1.14 vs. 3.9+/-0.97, respectively, p<0.05). Yet the mizoribine alone group did not showed any significant change of the DAS28 (4.3+/-0.56 vs. 3.9+/-0.37, respectively, p=0.076). Mild gastrointestinal disturbance was the most common adverse effect. The incidence of adverse effects was similar in both treatment groups (20% vs. 27%, respectively). CONCLUSIONS: Mizoribine in combination with MTX was effective for RA patients who showed an ineffective response or intolerance to MTX. Furthermore, this treatment can be considered to be relatively safe.
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Humans , Arthritis, Rheumatoid , Blood Cell Count , Creatinine , Incidence , Joints , Liver , Methotrexate , Pain Measurement , Retrospective StudiesABSTRACT
BACKGROUND: Mizoribine (MZR) is an imidazole nucleoside isolated from Eupenicillium brefeldianum. MZR is currently in clinical use for patients who have undergone renal transplantation. Therapeutic efficacy of MZR has also been demonstrated in rheumatoid arthritis and lupus nephritis. MZR has been shown to inhibit the proliferation of lymphocytes by interfering with inosine monophosphate dehydrogenase. Since the exact mechanism by which MZR benefits rheumatoid arthritis (RA) is not clear, we investigated the ability of MZR to direct its immunosuppressive influences on other antigen presenting cells, such as macrophages. METHODS: Mouse macrophage RAW264.7 cells were stimulated with lipopolysaccharide in the presence of MZR. To elucidate the mechanism of the therapeutic efficacy in chronic inflammatory diseases, we examined the effects of MZR on the production of pro-inflammatory cytokines, nitric oxide (NO) and prostaglandin E2 (PGE2) in macrophages. RESULTS: MZR dose-dependently decreased the production of nitric oxide and pro- inflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha), interleukins 1beta (IL-beta) and IL-6 PGE2. Examination of gene expression levels showed that the anti-inflammatory effect correlated with the down-regulation of inducible nitiric oxide synthase expression, cycloxygenase-2 expression and TNF-alpha gene expression. CONCLUSION: In this work, we resulted whether MZR (1.25~10 microgram/ml) inhibited macrophage activation by inhibiting secretion of pro-inflammatory cytokines, NO and PGE2. These findings provide an explanation for the therapeutic efficacy of MZR in chronic inflammation- associated diseases.
Subject(s)
Animals , Humans , Mice , Antigen-Presenting Cells , Arthritis, Rheumatoid , Cytokines , Dinoprostone , Down-Regulation , Eupenicillium , Gene Expression , Inosine Monophosphate , Interleukin-6 , Interleukins , Kidney Transplantation , Lupus Nephritis , Lymphocytes , Macrophage Activation , Macrophages , Nitric Oxide , Oxidoreductases , Tumor Necrosis Factor-alphaABSTRACT
PURPOSE: Mizoribine (MZR), an inhibitor of Inosine monophosphate (IMP) dehydrogenase which depletes cellular GTP, is clinically used as an immunosuppressive drug. This study was designed to evaluate the mechanism by which MZR exerts the cytotoxic effect on Jurkat T cells. METHODS: Jurkat T cell is a human T lymphocytic cell line. It was obtained from the Korean Type Culture Collection. Cell viability was measured by the MTT assay and flow cytometry. Caspase activity assay, Western blotting, 2-D PAGE, and mitochondrial membrane potential were detected using biochemical analysis. Morphologic finding was observed by Hoechst staining. RESULTS: The data demonstrated that the treatment of MZR decreased cell viability in a dose- and time-dependent manner. MZR-induced cell death was confirmed as apoptosis, which was characterized by chromatin condensation and H2AX phosphorylation. MZR increased the catalytic activity of caspase-3 protease, -8 protease and -9 proteases. The activation of caspase-3 protease was further confirmed by the degradation of polymerase (PARP), a substrate of caspase-3 protease by MZR in Jurkat T cells. Furthermore, MZR induced the changes of the mitochondrial transmembrane potential (MTP) and the cytosolic release of cytochrome c from the mitochondria. In addition, MZR induced the decrease of Bcl-X(L) expression whereas the increase of Bcl-X(S), Bak and Bim expression. Guanosine markedly inhibited cell viability and apoptosis through consistent suppression of the activity of caspase-8 protease, an upstream caspase among the caspase family, H2AX phosphorylation and PARP cleavage in MZR-treated cells. Also, I have screened the expression profile of proteins in the Jurkat T cells by using two-dimensional (2-D) gel electrophoresis. Among 300 spots resolved in the 2-D gels, the comparison of the control versus apoptotic cells revealed that the signal intensity of 10 spots was decreased and 5 spots was increased. CONCLUSION: The results suggest that MZR functions as an inhibitor of IMP dehydrogenase in apoptosis of Jurkat T cells via activation of intrinsic caspase cascades as well as mitochondrial dysfunction.
Subject(s)
Humans , Apoptosis , Blotting, Western , Caspase 3 , Caspase 8 , Cell Death , Cell Line , Cell Survival , Chromatin , Cytochromes c , Cytosol , Electrophoresis , Flow Cytometry , Gels , Guanosine , Guanosine Triphosphate , IMP Dehydrogenase , Inosine Monophosphate , Membrane Potential, Mitochondrial , Membrane Potentials , Mitochondria , Oxidoreductases , Peptide Hydrolases , Phosphorylation , T-LymphocytesABSTRACT
PURPOSE: Mizoribine (MZR), an inhibitor of Inosine monophosphate (IMP) dehydrogenase which depletes cellular GTP, is clinically used as an immunosuppressive drug. This study was designed to evaluate the mechanism by which MZR exerts the cytotoxic effect on Jurkat T cells. METHODS: Jurkat T cell is a human T lymphocytic cell line. It was obtained from the Korean Type Culture Collection. Cell viability was measured by the MTT assay and flow cytometry. Caspase activity assay, Western blotting, 2-D PAGE, and mitochondrial membrane potential were detected using biochemical analysis. Morphologic finding was observed by Hoechst staining. RESULTS: The data demonstrated that the treatment of MZR decreased cell viability in a dose- and time-dependent manner. MZR-induced cell death was confirmed as apoptosis, which was characterized by chromatin condensation and H2AX phosphorylation. MZR increased the catalytic activity of caspase-3 protease, -8 protease and -9 proteases. The activation of caspase-3 protease was further confirmed by the degradation of polymerase (PARP), a substrate of caspase-3 protease by MZR in Jurkat T cells. Furthermore, MZR induced the changes of the mitochondrial transmembrane potential (MTP) and the cytosolic release of cytochrome c from the mitochondria. In addition, MZR induced the decrease of Bcl-X(L) expression whereas the increase of Bcl-X(S), Bak and Bim expression. Guanosine markedly inhibited cell viability and apoptosis through consistent suppression of the activity of caspase-8 protease, an upstream caspase among the caspase family, H2AX phosphorylation and PARP cleavage in MZR-treated cells. Also, I have screened the expression profile of proteins in the Jurkat T cells by using two-dimensional (2-D) gel electrophoresis. Among 300 spots resolved in the 2-D gels, the comparison of the control versus apoptotic cells revealed that the signal intensity of 10 spots was decreased and 5 spots was increased. CONCLUSION: The results suggest that MZR functions as an inhibitor of IMP dehydrogenase in apoptosis of Jurkat T cells via activation of intrinsic caspase cascades as well as mitochondrial dysfunction.