Animals ; Brain ; metabolism ; Humans ; Nerve Regeneration ; drug effects ; Neurodegenerative Diseases ; metabolism ; prevention & control ; Neuroprotective Agents ; metabolism ; therapeutic use ; Tacrolimus Binding Protein 1A ; metabolism ; Tacrolimus Binding Proteins ; metabolism ; therapeutic use

AIMTo investigate the significance of the calcineurin (CaN) activation in active lupus nephritis patient.

METHODSPeripheral blood mononuclear cells (PBMCs) were separated from twenty-one active LN patients and 12 healthy controls. Phosphatase activity of CaN was determined using the CaN assay kit by measuring the content of released PO4. Reverse transcription-PCR was used to detect the expression of CD40L mRNA. Flow cytometry analysis was used to detect the expression of CD40L in LN PBMC.

RESULTS(1) Increased activation of CaN in spontaneous cultured PBMC in active LN group was found as compared with control group (46.08 +/- 5.58 vs 8.81 +/- 3.61, P < 0.01). In stimulated by PMA/Ionomycin , activity of CaN in active LN group was also higher than that of control (69.34 +/- 12.59 vs 37.12 +/- 11.57, P < 0.01). (2) Relative content of CD40L in PBMC in active LN groups increased significantly as compared with the control groups under spontaneous and PMA/Ionomycin-induced culture, respectively (P < 0.01). (3) FK506 reduced significantly production of CD40L in spontaneous and PMA/Ionomycin-induced PBMC of LN.

CONCLUSIONElevated activation of CaN in active LN may participate in regulation overexpression of CD40L in PBMC of LN. Through inhibiting CaN activity, FK506 may prevent abnormal activation of CD40-CD40L costimulatory pathway in lupus nephritis.

Adolescent ; Adult ; CD40 Ligand ; metabolism ; Calcineurin ; metabolism ; Case-Control Studies ; Cells, Cultured ; Female ; Humans ; Leukocytes, Mononuclear ; metabolism ; Lupus Nephritis ; blood ; metabolism ; Male ; Middle Aged ; Tacrolimus ; pharmacology ; Young Adult

OBJECTIVETo investigate the mechanism underlying the inhibitory effect of tacrolimus (FK506) against acute liver graft rejection.

METHODSRat models of orthotopic liver transplantation were divided into 3 groups, namely the tolerance group with Brown Norway (BN) rats as the donors and Lewis rats as the recipients, rejection group with Lewis rats as donors and BN rats as recipients, and FK506 group with the same donor-recipient pair as in the rejection group and FK506 treatment. The recipients were sacrificed 7 days after the transplantation, and the hepatic histology, cytokine levels, and glucocorticoid-induced tumor necrosis factor-related protein ligand (GITRL) expression in the liver and Kupffer cells were observed and detected.

RESULTSCompared with the tolerance group, the rejection group showed increased GITRL expressions in the liver and Kupffer cells (P<0.05), which was significantly lowered by FK506 treatment (P<0.05). Acute liver graft rejection caused significantly elevated interferon-γ (IFN-γ) levels and decreased interleukin-10 (IL-10) levels in the plasma and Kupffer cells (P<0.05), and these changes were obviously attenuated by FK506 treatment (P<0.05).

CONCLUSIONThe effect of FK506 in suppressing acute liver graft rejection is probably associated with down-regulated GITRL expression in the liver and Kupffer cells.

Animals ; Carrier Proteins ; metabolism ; Graft Rejection ; prevention & control ; Kupffer Cells ; metabolism ; Liver ; metabolism ; Liver Transplantation ; Male ; Rats ; Rats, Inbred BN ; Rats, Inbred Lew ; Tacrolimus ; pharmacology

OBJECTIVETo explore the effect of tacrolimus on murine hair follicle cycle.

METHODHematoxylin-eosin dyeing and reverse transcription-polymerase chain raction techniques were used.

RESULTSFive days after depilation, the hair follicles in both the tacrolimus group and the minoxidil group was in anagen V, while that in the vaseline group was in anagen III. vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) were detected in back skin in both the tacrolimus group and the minoxidil group, but not in the vaseline group.

CONCLUSIONTacrolimus can promote the growth of hair by stimulating the hair follicle to enter anagen V in mice, which may be explained by the effects of VEGF and HGF.

Animals ; Hair Follicle ; drug effects ; physiology ; Hepatocyte Growth Factor ; metabolism ; Mice ; Mice, Inbred C57BL ; Minoxidil ; pharmacology ; Skin ; drug effects ; metabolism ; Tacrolimus ; pharmacology ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo observe the change of Th immunological gene in renal transplant recipients after the treatment of cyclosporine (CsA) and tacrolimus (FK506).

METHODSThe peripheral blood lymphacytes just before and 24 hours after CsA and FK506 treatment were isolated. The total RNA of them were reverse-transcripted and examined by real-time quantity PCR array. The results were analyzed by bioinformatic methods.

RESULTSThe TLR4, CEBPB, IL4R, IL1R1,IL18R1,and IL1R2 genes were remarkably upregulated, whereas IL-2, CCL5, CD27, CCR5, CCR4, CD4, RPL13A, TGFB3, CD86, CCR3, STAT1, NFATC2IP, IL23A, IL15, IRF4, and TFCP2 were downregulated 24 hours after CsA treatment. The IL18, IL7, PTPRC, TNFSF4, SPP1, GFI1, TLR4, IL13RA1, TNF, INHBA, LAG3, IL13, IL1R1, SOCS5, IL10, YY1, TBX21, FASLG, IL18R1, and IL1R2 genes were remarkably upregulated, whereas IL-2, IL-3, IL-4, IL-6,CCR5, CD4, CD27, CD40LG, IL15, CCR3, CD86, CCR4, and IRF4 were obviously downregulated 24 hours after FK506 treatment.

CONCLUSIONCsA and FK506 exert their therapeutic effectiveness by regulating the expressions of a series of target genes.

Cyclosporine ; pharmacology ; Cytokines ; genetics ; metabolism ; Gene Expression Regulation ; Humans ; Kidney ; drug effects ; metabolism ; Kidney Transplantation ; Oligonucleotide Array Sequence Analysis ; T-Lymphocytes, Helper-Inducer ; drug effects ; metabolism ; Tacrolimus ; pharmacology

Two intracellular signal pathways mediated by cAMP and protein kinase C (PKC) were involved in the regulation of FN gene expression (Lee et al., Exp. Mol. Med. 30: 240, 1998). In this study, a possible involvement of protein phosphatase-dependent pathways in the regulation of FN gene expression was investigated by using protein phosphatase type 2B (PP2B) inhibitors, cyclosporin A and ascomycin. Both cyclosporin A and ascomycin increased the levels of FN mRNA in WI-38 human lung fibroblasts and the SV40-transformed WI-38 cells but not in MC3T3-E1 osteoblasts. The expression of FN appears to increase from six hours up to 48 hours after treatment suggesting that it is not an immediate effect. In addition, this effect required a new protein synthesis. Neither cyclosporin A nor ascomycin affects the phorbol myristate acetate (PMA)-induced stimulation of FN gene expression and the same result occurred in vice versa suggesting the mechanism of PMA and cyclosporin A/ascomycin in the regulation of FN gene expression may share a common downstream pathway. Taken together, this study suggests that PP2B is involved in the regulation of FN gene expression in normal and transformed fibroblasts but not in osteoblasts.
Animal ; Calcineurin/antagonists & inhibitors* ; Cell Line, Transformed ; Cell Transformation, Viral ; Cyclosporine/pharmacology* ; Enzyme Inhibitors/pharmacology ; Fibroblasts ; Fibronectins/metabolism ; Fibronectins/genetics* ; Gene Expression Regulation* ; Human ; Lung/cytology ; Mice ; Osteoblasts ; Tacrolimus/pharmacology ; Tacrolimus/analogs & derivatives*

This study is to investigate the effect of FK506 on expression of hepatocyte growth factor (HGF) in rats' spinal cord following peripheral nerve injury and to elucidate the mechanisms for neuroprotective property of FK506. Fifty male rats were randomly divided into normal group, injury group and treatment group. Models of peripheral nerve injury were established by bilateral transection of sciatic nerve 0.5 cm distal to piriform muscle. Then the treatment group received subcutaneous injection of FK506 (1 mg/kg) at the back of neck, while the injury group was given 0.9% saline. The L(4-6) spinal cords were harvested at various time points after the surgery. Western blotting and immunofluorescent staining were used to detect the level and position of HGF in spinal cord. Immunofluorescent staining showed that HGF-positive neurons were located in anterior horn, intermediate zone and posterior horn of gray matter in normal spinal cord. Western blotting revealed that there was no significant difference in the expressions of HGF between the injury group and the normal group, while the expression of HGF was significantly higher in the treatment group than in the injury group 7 and 14 days after surgery. It is suggested that peripheral nerve injury does not result in up-regulation of the expression of HGF in spinal cord, while FK506 may induce high expression of endogenous HGF after injury thereby protecting neurons and promoting axonal outgrowth.
Cells, Cultured ; Gene Expression Regulation ; Hepatocyte Growth Factor/metabolism ; Immunosuppressive Agents/metabolism ; Immunosuppressive Agents/*pharmacology ; Microscopy, Fluorescence/methods ; Neurons/metabolism ; Peripheral Nervous System/*metabolism ; Sciatic Nerve/metabolism ; Spinal Cord/*cytology ; Spinal Cord/metabolism ; Spinal Cord Injuries/*drug therapy ; Tacrolimus/metabolism ; Tacrolimus/*pharmacology

PURPOSE: Liver transplantation (LT) can cure abnormality of glucose metabolism, but cause altered glucose metabolism with immunosuppressive treatment. Up to now, almost all studies have been performed in cadaveric donor liver transplantation (CDLT). We underwent study in CDLT and also living donor liver transplantation (LDLT) recipients. METHODS: Among 397 adult-to-adult LT recipients between January 1994 and August 2001, we selected 81 patients who could be followed more than 12 months by using the table of random sampling numbers. We reviewed the change of blood glucose and risk factors, complications and survival retrospectively between post-transplantation diabetes mellitus (PTDM) and no PTDM patients. RESULTS: Clinical data showed 34 : 47 in frequency of PTDM to no PTDM. Age, family history of DM, preoperative DM history over 6 months had a significant risk of PTDM. There was no difference of PTDM frequency between CDLT and LDLT and its subgroup. The worse post-transplant graft function causes the more incidence of PTDM (P=0.051). FK506 had higher relation with PTDM than cyclosporine and mycophenolate mofetile (P=0.058). The incidence of DM after operation has been decreased by 6 months, but thereafter no further. There were 18 of De Novo DM among 34 PTDM patients, and only 1 preoperative DM patient improved after LT. Between PTDM and no PTDM group, there were no significant difference of complication rate and 5-year survival rate. CONCLUSIONS: The types of graft would not affect the incidence of PTDM if the graft function were preserved. Other clinical data showed similar results to previous reports.
Blood Glucose ; Cadaver ; Cyclosporine ; Diabetes Mellitus* ; Glucose ; Humans ; Incidence ; Liver Transplantation* ; Liver* ; Living Donors ; Metabolism ; Retrospective Studies ; Risk Factors ; Survival Rate ; Tacrolimus ; Tissue Donors ; Transplants

Posttransplant diabetes mellitus, a complication due to corticosteroids and the calcineurin inhibitors, cyclosporine and tacrolimus, is commonly regarded as a form of type 2 diabetes mellitus. Diabetes ketoacidosis, which requires relative insulin deficiency to impair fatty acid metabolism, is a complication of type 1 diabetes mellitus. We report two patients who presented with diabetic ketoacidosis after kidney transplantation. Two patients presented with severe hyperglycemia, significant ketosis and metabolic acidosis of variable severity. One patient was treated with a cyclosporine-based regimen, and the other with a tacrolimus-based regimen. Both were found to have moderate to high serum levels of calcineurin inhibitors on presentation. Our experience suggests that post-transplant diabetes mellitus, in association with calcineurin inhibitor, may result in ketoacidosis either secondary to relative beta cell dysfunction, peripheral insulin resistance, or a combination of the two effects. Post transplant diabetes mellitus can be an atypical form of adult-onset diabetes with features of both type 1 and type 2 diabetes mellitus.
Acidosis ; Adrenal Cortex Hormones ; Calcineurin ; Cyclosporine ; Diabetes Mellitus ; Diabetes Mellitus, Type 1 ; Diabetes Mellitus, Type 2 ; Diabetic Ketoacidosis ; Humans ; Hyperglycemia ; Insulin ; Insulin Resistance ; Ketosis ; Kidney Transplantation ; Metabolism ; Tacrolimus

Posttransplant diabetes mellitus, a complication due to corticosteroids and the calcineurin inhibitors, cyclosporine and tacrolimus, is commonly regarded as a form of type 2 diabetes mellitus. Diabetes ketoacidosis, which requires relative insulin deficiency to impair fatty acid metabolism, is a complication of type 1 diabetes mellitus. We report two patients who presented with diabetic ketoacidosis after kidney transplantation. Two patients presented with severe hyperglycemia, significant ketosis and metabolic acidosis of variable severity. One patient was treated with a cyclosporine-based regimen, and the other with a tacrolimus-based regimen. Both were found to have moderate to high serum levels of calcineurin inhibitors on presentation. Our experience suggests that post-transplant diabetes mellitus, in association with calcineurin inhibitor, may result in ketoacidosis either secondary to relative beta cell dysfunction, peripheral insulin resistance, or a combination of the two effects. Post transplant diabetes mellitus can be an atypical form of adult-onset diabetes with features of both type 1 and type 2 diabetes mellitus.
Acidosis ; Adrenal Cortex Hormones ; Calcineurin ; Cyclosporine ; Diabetes Mellitus ; Diabetes Mellitus, Type 1 ; Diabetes Mellitus, Type 2 ; Diabetic Ketoacidosis ; Humans ; Hyperglycemia ; Insulin ; Insulin Resistance ; Ketosis ; Kidney Transplantation ; Metabolism ; Tacrolimus