To investigate the effects of mycophenolate mofetil (MMF) on the process of renal interstitial fibrosis, unilateral ureteral obstruction (UUO) model was established in rats. Twenty Sprague-Dawley rats underwent UUO and received vehicle (n = 10) or MMF (20 mg.kg-1.d-1, by daily gastric gavage, n = 10) during a period of 5 days following surgery, and the additional 10 rats were served as sham-operated group. The rats were killed 5 days after surgery. Immunohistochemistry was performed on renal tissue for proliferating cell nuclear antigen (PCNA), alpha-smooth muscle actin (alpha-SMA) and type I and III collagen (col I, col III). Histological studies were also done by MASSON staining. Five days after surgery, proliferating cells in tubules, interstitium as well as interstitial myofibroblast (MyoF) infiltration and interstitial col I, col III deposition were all significantly reduced by MMF treatment. MMF also alleviated the histological changes of UUO rats. These results suggested that the reduction of interstitial MyoF infiltration may be an important event by which MMF prevents renal injury caused by UUO and MMF could be used to limit the progression of renal fibrosis.
Fibrosis ; Kidney/*pathology ; Kidney Diseases/etiology ; Kidney Diseases/pathology ; Kidney Diseases/*prevention & control ; Mycophenolic Acid/*analogs & derivatives ; Mycophenolic Acid/*pharmacology ; Random Allocation ; Rats, Sprague-Dawley ; Ureteral Obstruction/*complications

BackgroundThe previous study showed that mycophenolic acid (MPA) synergizing with lipopolysaccharide (LPS) promoted interleukin (IL)-1β release, but the mechanism is unclear. This study aimed to investigate the mechanism of MPA synergizing with LPS to induce IL-1β release.

MethodsUndiluted human blood cells, THP-1 human myeloid leukemia mononuclear cells (THP-1) cells, or monocytes were stimulated with LPS and treated with or without MPA, and the supernatant IL-1β was detected by enzyme-linked immunosorbent assay. The mRNA levels of IL-1β were detected by real-time quantitative polymerase chain reaction. The intracellular protein levels of nuclear factor kappa B (NF-κB) phospho-p65 (p-p65), precursor interleukin-1β (pro-IL-1β), NOD-like receptor pyrin domain containing-3 (NLRP3), and cysteine aspartic acid-specific protease-1 (caspase-1) p20 in THP-1 cell were measured by Western blot.

ResultsThe MPA alone failed to induce IL-1β, whereas MPA synergized with LPS to increase IL-1β in a dose-dependent manner (685.00 ± 20.00 pg/ml in LPS + 5 μmol/L MPA group, P = 0.035; 742.00 ± 31.58 pg/ml in LPS + 25 μmol/L MPA group, P = 0.017; 1000.00 ± 65.59 pg/ml in LPS + 75 μmol/L MPA group, P = 0.024; versus 408.00 ± 35.50 pg/ml in LPS group). MPA alone has no effect on the IL-1β mRNA expression, LPS induced the expression of IL-1β mRNA 2761 fold, and LPS + MPA increased the IL-1β expression 3018 fold, which had the same effect with LPS group (P = 0.834). MPA did not affect the intracellular NF-κB p-p65 and pro-IL-1β protein levels but activated NLRP3 inflammasome. Ac-YVAD-cmk blocked the activation of caspase-1 and subsequently attenuated IL-1β secretion (181.00 ± 45.24 pg/ml in LPS + MPA + YVAD group vs. 588.00 ± 41.99 pg/ml in LPS + MPA group, P = 0.014).

ConclusionsTaken together, MPA synergized with LPS to induce IL-1β release via the activation of caspase-1, rather than the enhanced production of pro-IL-1β. These findings suggested that patients immunosuppressed with mycophenolate mofetil may have overly activated caspase-1 during infection, which might contribute to a more sensitive host defense response to invading germs.

Animals ; Caspase 1 ; metabolism ; Cells, Cultured ; Humans ; Inflammasomes ; Interleukin-1beta ; metabolism ; Lipopolysaccharides ; pharmacology ; Mice ; Mice, Inbred NOD ; Mycophenolic Acid ; pharmacology ; NLR Family, Pyrin Domain-Containing 3 Protein

OBJECTIVETo determine whether mycophenolic acid (MPA) exerts an apoptotic effect on human leukemic T cells Molt-4 and to elucidate its possible mechanism.

METHODSCell morphology, DNA fragmentation, cell cycle,percentage of annexin V positive cells and enzymatic activity of caspase-3 were measured by microscopic, electrophoretic and flow cytometric techniques respectively. Human leukemic B cell line Raji was used as control.

RESULTMPA could induce apoptosis of Molt-4 cells with dose-and time-dependent manners; cells were blocked in the S phase. The activity of caspase-3 was enhanced in a dose-dependent manner in Molt-4 cells treated with MPA for 24 h. MPA could not induce apoptosis in Raji cells.

CONCLUSIONMPA can induce apoptosis in Molt-4 cells which may be involved in the activation of caspase-3.

Antibiotics, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; Caspases ; metabolism ; Dose-Response Relationship, Drug ; Humans ; Leukemia, T-Cell ; pathology ; Mycophenolic Acid ; pharmacology ; Time Factors ; Tumor Cells, Cultured

To investigate the effects of mycophenolate mofetil (MMF) on the process of renal interstitial fibrosis, unilateral ureteral obstruction (UUO) model was established in rats. Twenty Sprague-Dawley rats underwent UUO and received vehicle (n = 10) or MMF (20 mg.kg-1.d-1, by daily gastric gavage, n = 10) during a period of 5 days following surgery, and the additional 10 rats were served as sham-operated group. The rats were killed 5 days after surgery. Immunohistochemistry was performed on renal tissue for proliferating cell nuclear antigen (PCNA), alpha-smooth muscle actin (alpha-SMA) and type I and III collagen (col I, col III). Histological studies were also done by MASSON staining. Five days after surgery, proliferating cells in tubules, interstitium as well as interstitial myofibroblast (MyoF) infiltration and interstitial col I, col III deposition were all significantly reduced by MMF treatment. MMF also alleviated the histological changes of UUO rats. These results suggested that the reduction of interstitial MyoF infiltration may be an important event by which MMF prevents renal injury caused by UUO and MMF could be used to limit the progression of renal fibrosis.
Animals ; Female ; Fibrosis ; Kidney ; pathology ; Kidney Diseases ; etiology ; pathology ; prevention & control ; Mycophenolic Acid ; analogs & derivatives ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Ureteral Obstruction ; complications

OBJECTIVETo study the effects of different immunodepressants on the sperm parameters of kidney transplant recipients.

METHODSIn 15 healthy fertile men and 37 kidney transplant recipients, ejaculates were aseptically obtained by masturbation. Thirty-seven patients were divided into two groups, 20 patients were treated with Prograf (FK506) combination with mycophenolate mofetil (MMF) and prednisone; 17 patients were treated with cyclosporine (CsA) combination with azathioprine with prednisone. The sperm viability, mobility parameters such as prorsad percentage motility, straight line velocity (VSL), curve line velocity (VCL), velocity of average path (VAP) and morph were estimated with a computer-assisted sperm analyzer (CASA) provided with a multiple-exposure photography system.

RESULTSThere were no significant difference in sperm viability rate [(81.7 +/- 5.7)%, (79.4 +/- 6.8)% and (83.8 +/- 6.0)%], VCL [(24.1 +/- 8.6)%, (23.9 +/- 4.4)%, (24.8 +/- 4.2)% ] and VAP [(19.7 +/- 6.6)%, (18.6 +/- 2.9)%, (21.0 +/- 4.0)%] among groups of FK506, CsA and control, respectively (P > 0.05). The rate of anomaly [(67.8 +/- 5.7)%], the prorsad percentage motility [(46.4 +/- 8.1)%] and VSL [(15.4 +/- 4.6)%] in the group of FK506 were respectively significantly lower and higher than those in the group of CsA [(80.1 +/- 5.6%, (33.3 +/- 6.4)%, (10.2 +/- 2.4)%] (P < 0.05).

CONCLUSIONThe application of FK506 combined with MMF could help recover the mobility and morphology of the sperm in kidney transplantation recipients.

Adolescent ; Adult ; Case-Control Studies ; Cyclosporine ; pharmacology ; Drug Therapy, Combination ; Humans ; Immunosuppressive Agents ; pharmacology ; Kidney Transplantation ; Male ; Mycophenolic Acid ; analogs & derivatives ; pharmacology ; Prednisone ; pharmacology ; Sperm Motility ; drug effects ; Tacrolimus ; pharmacology

OBJECTIVEIt is well known that cyclosporine A (CsA), a widely used immunosuppressant for clinical organ transplantation, has the ability to inhibit HCV replication. In this study, the effects of several other immunosuppressants, including mycophenolic acid (MPA), rapamycin and FK-506, on HCV replication were examined in human hepatocytes.

METHODSHCV JFH-l-infected hepatocytes were treated with immunosuppressants or with control vehicles. The levels of viral RNA and the expression of HCV core protein were determined by quantitative real-time RT-PCR and Western Blot assay, respectively.

RESULTSMPA-treated cells showed significant decreases in both viral RNA and HCV Core protein expression compared with the control cells. Moreover, MPA treatments of hepatocytes before, during or after HCV infection could significantly inhibit viral replication. In contrast, rapamycin and FK-506 had little effect on HCV replication. Mechanism research disclosed that the inhibition of HCV replication by MPA was mainly due to its depletion of guanosine, a purine nucleoside crucial for synthesis of guanosine triphosphate (GTP), which is required for initiation of HCV RNA replication. The supplement of exogenous guanosine could reverse most of anti-HCV effect of MPA.

CONCLUSIONThese results indicate that MPA, through the depletion of guanosine, inhibits HCV JFH-1 replication in hepatocytes, suggesting that MPA may be beneficial for HCV-infected transplant recipients.

Cell Line, Tumor ; Hepacivirus ; drug effects ; genetics ; physiology ; Hepatitis C ; drug therapy ; virology ; Hepatocytes ; virology ; Humans ; Immunosuppressive Agents ; pharmacology ; Mycophenolic Acid ; pharmacology ; RNA, Viral ; genetics ; Virus Replication ; drug effects ; genetics

AIM: To investigate the chemical constituents of the cultures of Laetiporus sulphureus (Bull.) Murrill. METHOD: Compounds were isolated and purified by various chromatographic techniques. The structure of the new compound was determined by interpretation of MS and 1D-, 2D-NMR spectroscopic data, while the known compounds were identified by comparison of their data with those reported. RESULTS: Three mycophenolic acid derivatives, 6-((2E, 6E)-3, 7-dimethyldeca-2, 6-dienyl)-7-hydroxy-5-methoxy-4-methylphtanlan-1-one (1), 6-((2E, 6E)-3, 7, 11-trimethyldedoca-2, 6, 10-trienyl)-5, 7-dihydroxy-4-methylphtanlan-1-one (2), and 6-((2E, 6E)-3, 7, 11-trimethyldedoca-2, 6, 10-trienyl)-7-hydroxy-5-methoxy-4-methylphtanlan-1-one (3) were isolated. CONCLUSION Among them, compound 1 was new, and compound 2 exhibited moderate cytotoxicity against HL-60, SMMC-7721, A-549, and MCF-7 cells, with IC50 values of 39.1, 31.1, 27.4, and 35.7 μmol·L(-1), respectively.
Agaricales ; Biological Products ; chemistry ; isolation & purification ; pharmacology ; therapeutic use ; HL-60 Cells ; Humans ; MCF-7 Cells ; Molecular Structure ; Mycophenolic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; Neoplasms ; drug therapy ; Phenols ; chemistry ; isolation & purification ; pharmacology ; therapeutic use ; Polyporales ; chemistry

OBJECTIVETo evaluate the effect of mycophenolate mofetile (MMF) on acute graft versus host disease (aGVHD) prophylaxis after allogeneic bone marrow transplantation (allo-BMT) in a murine model.

METHODSAcute GVHD was induced in male BALB/c mice by total-body irradiation (TBI) followed by female allogeneic (C57BL/6J) bone marrow and spleen cells transplantation. Acute GVHD was assessed both physically and histologically. Severe aGVHD was developed in the recipients and the mean survival time (MST) of untreated BM recipients was 6 days. After allo-BMT, animals were divided into 6 groups. Group 1 was given MMF (30 mg.kg(-1).d(-1)), group 2 CsA (1.5 mg.kg(-1).d(-1)) and MTX (0.4 mg.kg(-1).d(-1)), group 3 MMF (30 mg.kg(-1).d(-1)) in combination with CsA and MTX, group 4 MMF (60 mg.kg(-1).d(-1)) in combination with CsA and MTX, group 5 MMF (10 mg.kg(-1).d(-1)) in combination with CsA and MTX, and group 6 no agents for aGVHD prophylaxis. The physical signs, MST, peripheral blood counts, and aGVHD histopathologic examination were observed in all recipients.

RESULTSAnimals in control group developed typical aGVHD and 100% of mortality, with a MST of 6 days. The MSTs of group 1 approximately 5 were significantly longer than that of control, being 3.4, 8.4, 9.0, 6.1 and 8.8 days longer, respectively (P < 0.05). The MSTs of groups 2 approximately 5 were longer than that of group 1 (P < 0.05), but there was no statistical significance between groups 3 approximately 5 and 2. There was no statistical difference in peripheral blood count among groups 1 approximately 5. Histopathological examination of skin, liver, and gastrointestinal tract showed typical signs of aGVHD. Animals received immunosuppressive agents (MMF, CsA, MTX) showed the less severe signs.

CONCLUSIONSMMF markedly prolonged MST of allo-BMT recipients, delayed the onset of aGVHD signs. The prophylaxis effect of CsA + MTX with or without MMF was better than that of MMF alone, synergism between MMF of 10 or 30 mg.kg(-1).d(-1) and CsA + MTX was better than that of MMF of 60 mg.kg(-1).d(-1) and CsA + MTX.

Animals ; Bone Marrow Transplantation ; Cyclosporine ; pharmacology ; Dose-Response Relationship, Drug ; Drug Synergism ; Female ; Graft Survival ; drug effects ; Graft vs Host Disease ; pathology ; prevention & control ; Immunosuppressive Agents ; pharmacology ; Methotrexate ; pharmacology ; Mice ; Mice, Inbred C57BL ; Models, Animal ; Mycophenolic Acid ; analogs & derivatives ; pharmacology ; Time Factors ; Transplantation, Homologous

We performed this study to investigate the feature of rejection in porcine-to-rat corneal orthotopic transplantation and to evaluate the effect of cyclosporine and mycophenolate on the xeno-rejection. Orthotopic corneal transplantation was done at 91 Sprague-Dawley rats, and they were divided into 10 groups based on the combination of immunosuppressants including dexamethasone, cyclosporine, and mycophenolate mofetil. Graft survival was analyzed and grafted eyes were examined with Hematoxylin & Eosin and CD4 or CD8 staining. Enzyme-linked immunosorbent assays were done for interleukin-2 (IL-2), IL-4, IL-5, IL-10, and interferon (IFN)-gamma in cornea, lacrimal gland, and cervical lymph nodes. The longest median survival of the immune suppressant group was 11.00+/-1.96 days, which showed no statistical differences compared with that of control (8.00+/-1.52 days). The neutrophils were prominent in the early phase but soon gave way to the monocytes. The number of CD8+ cells was higher than that of CD4+ cells. IL-2 and IFN-gamma markedly increased at 10 to13 days in cornea, lacrimal glands, and cervical lymph nodes, which showed a decrease with immunosuppressants except in the cornea. In conclusion, cyclosporine and mycophenolate could not prevent the rejection in porcine to rat orthotopic corneal xenograft associated with infiltraton of CD8+ and innate immune cells.
Animals ; *Corneal Transplantation ; Cyclosporine/*pharmacology ; Cytokines/metabolism ; Graft Rejection/immunology/pathology/*prevention & control ; Graft Survival/*drug effects ; Immunosuppressive Agents/*pharmacology ; Interferon-gamma/metabolism ; Interleukin-10/metabolism ; Interleukin-2/metabolism ; Interleukin-4/metabolism ; Interleukin-5/metabolism ; Mycophenolic Acid/*analogs & derivatives/pharmacology ; Neutrophils/immunology ; Rats ; Rats, Sprague-Dawley ; Swine ; Transplantation, Heterologous

In order to evaluate whether immunosuppressive agents such as mycophenolate mofetil (MMF) and azathioprine would differently influence the outcome of the renal transplants, we prospectively analyzed the incidence of acute rejection episodes, cytomegalovirus infection within the first 6 months following renal transplantation and 5 yr graft survival rate after minimizing influences of donor factors by grafting the same cadaveric donor kidney. There was no significant difference in sex, HLA mismatch, cold ischemic time, and patients' weight between the two groups. Contrary to the previous studies which demonstrated that MMF could lower the incidence of acute rejection episodes and improved graft survival rate, the two groups showed no significant difference in the incidence of acute rejection episodes and 5-yr graft survival rate as well. This discrepancy in these results might explain that donor factors could be important to cadaveric renal transplantation. Thus, we suggest that the influences of donor factors should be considered in further clinical studies of cadaveric renal transplantation.
ABO Blood-Group System ; Adult ; Azathioprine/*pharmacology ; Body Weight ; Cadaver ; Cytomegalovirus/metabolism ; Cytomegalovirus Infections/metabolism ; Female ; Graft Rejection ; Graft Survival ; Histocompatibility Testing ; Humans ; Immunophenotyping ; Immunosuppressive Agents/pharmacology ; Ischemia ; Kidney Diseases/mortality/therapy ; Kidney Transplantation/*methods ; Male ; Middle Aged ; Mycophenolic Acid/*analogs & derivatives/*pharmacology ; Prospective Studies ; Time Factors ; Tissue Donors ; Treatment Outcome