Cyclosporine A Group Research Articles

Cyclosporine A improves the binding of mouse embryos to fibronectin.

The binding of integrin αvβ3 with endometrial fibronectin (FN) promotes the migration of preimplantation embryos in mice. We have previously shown that cyclosporine A (CsA) improves the adhesion and invasion of mouse preimplantation embryos. In this study, we evaluated the roles of calcium ions and downstream signaling factors in the binding of integrin αvβ3 to FN. Female Institute of Cancer Research (ICR) mice were superovulated and mated, and two-cell embryos were harvested from the oviducts and cultured to the blastocyst stage The adhesion and stretching growth of hatched embryos in laminin-coated dishes were evaluated, and integrinβ3 expression was determined using qPCR. Blastocytes were cultured with 0 or 1 cyclosporine A (CsA) and the attachment of embryonic integrin αvβ3 to FN120 was observed using a fluorescent bead. To further determine the mechanism, the cells were also incubated with calcium ions and protein kinase C and calmodulin antagonists. The binding of integrin αvβ3 to FN120 was examined via confocal laser scanning microscopy. The adhesion and stretching growth of peri-implantation embryos were greater and integrinβ3 expression was higher in the 1 μM CsA group than in the 0 μM CsA group (p < 0.05). When incubated with calcium ions and protein kinase C and calmodulin antagonists, the ability of peri-implantation embryos to bind to FN decreased; CsA treatment promoted this binding. This study revealed that CsA up - regulates integrinβ3 expression in peri - implantation embryos and promotes binding to FN via calcium ions, and protein kinase C, and calmodulin. These findings provide evidence supporting the beneficial effect of CsA on the peri - implantation embryo adhesion.

The effect of intense pulsed light combined with topical 0.05% Cyclosporin A eyedrops in the treatment of Sjögren’s syndrome related dry eye

ABSTRACT Objectives This study aimed to assess the effectiveness and safety of intense pulsed light (IPL) therapy plus topical 0.05% cyclosporine A (CsA) eye drops to treat Sjögren’s Syndrome-related dry eyes (SS-DE). Research design and methods In this prospective, randomized trial included, 60 individuals with SS-DE symptoms were randomized to receive topical eye drops containing either 0.1% sodium hyaluronate (Group S) or 0.05% CsA (Group C) plus IPL therapy. Before the first treatment (baseline), and at 12, 16, and 20 weeks after treatment commencement, we assessed the best corrected visual acuity (BCVA), the Ocular Surface Disease Index (OSDI) score, the Schirmer I test (SIT), noninvasive tear breakup time (NBUT), corneal fluorescein staining (CFS), meibomian gland (MG) dropout, lid margin abnormality, MG expressibility, and meibum quality. Results Both groups showed significant improvements in the OSDI, NBUT, CFS, MG expressibility, and meibum quality (all p < 0.05). Group C showed a greater increase in OSDI, NBUT, MG expressibility, and meibum quality (all p < 0.05). Moreover, SIT and lid margin abnormalities significantly improved in Group C (both p < 0.05), but not in Group S. Conclusion Treatment with 0.05% CsA eyedrops plus IPL therapy could significantly reduce the issues and physical discomfort of patients with SS-DE. Clinical Trial Registered on 20 July 2021, with the registration number ChiCTR2100049059.

Comparison of tacrolimus vs. cyclosporine in pediatric hematopoietic stem cell transplantation for thalassemia.

Graft-versus-host disease (GvHD) is one of the leading causes of morbidity and mortality in patients undergoing allogeneic HSCT, and effective prevention of GvHD is critical for the success of the HSCT procedure. Calcineurin inhibitors (CNI) have been used for decades as the backbone of GvHD prophylaxis. In this study, the efficacy and safety of Cyclosporine A (CsA) and tacrolimus (TCR) were compared in pediatric HSCT for thalassemia. This is a retrospective analysis of 129 pediatric patients who underwent HSCT with the diagnosis of thalassemia at Medicalpark Göztepe and Antalya Hospitals between January 2017 and December 2020. Despite the GvHD prophylaxis, grade II-IV acute GvHD developed in 29 patients. Of these patients, 12 had only gut, 10 had only skin, 6 had combined gut and skin, and one had only liver GvHD. Fifteen of these 29 patients were in the CsA group, and 14 of them were in the TCR group. There was no significant difference between the groups in terms of acute GvHD occurrence, GvHD stage, or involvement sites. In terms of CNI-related toxicity, neurotoxicity in 15 (CsA n = 9, TCR n = 6) and nephrotoxicity in 18 (CsA n = 4, TCR n = 14) patients were observed. While there was no difference between the two groups in terms of neurotoxicity, more nephrotoxicity developed in patients using TCR (p = .013). There was no significant difference between the groups in terms of engraftment syndrome, veno-occlusive disease, CMV reactivation, PRES, or graft rejection. Regarding GvHD, there was no difference in efficacy between TCR and CsA usage. Patients taking TCR experienced noticeably higher nephrotoxicity in terms of adverse effects. This difference should be considered according to the patient's clinical situation while choosing a CNI.

Umbilical Cord Blood-Derived Cells Can Reconstruct Hematopoiesis in an Aplastic Anemia Animal Model.

To explore the efficacy and the mechanism of the umbilical cord-derived cells combined with cyclosporine A (CsA) in treating aplastic anemia (AA) in mice. Immune-mediated AA model mice were treated with CsA + UC mesenchymal stem cells (UC-MSC), CsA + umbilical cord blood regulatory T cells (UCB-Treg), UC-MSC, UCB-Treg, CsA alone, or blank control, respectively (n = 9 mice/group). CsA and the cell infusion was administered on d0. Routine peripheral blood testing was performed once weekly; bone marrow colony culture, bone marrow cell flow cytometry, peripheral blood T cell subsets, and serum inflammatory cytokines tests were performed on d14. Transcriptome sequencing was performed for cells from CsA + UC-MSC, CsA + UCB-Treg, and CsA groups to detect the possible related genes. Gene function cluster and signal pathway enrichment analysis were also performed. Blank control mice died due to pancytopenia within 21 days, whereas mice in other groups survived for >28 days. On d14, the CsA + UC-MSC and CsA + UCB-Treg groups had higher white blood cell (WBC) counts than the other groups (p < 0.05), along with higher burst-forming unit (BFU) and colony-forming unit-granulocyte, macrophage (CFU-GM) counts (p < 0.01). The CsA + UC-MSC group had the highest BFU count (p < 0.01). The CsA + UC-MSC and CsA + UCB-Treg groups exhibited the highest bone marrow CD34+ cell proportion (9.68% ± 1.35% and 8.17% ± 0.53%, respectively; p < 0.01). Tumor necrosis factor (TNF)-α and interleukin (IL)-2 levels in the CsA + UC-MSC group (p < 0.05) and TNF-α, interleukin-2, and interferon (INF)-γ levels in the CsA + UC-Treg group (p < 0.01) were lower than those in the CsA group. Compared with CsA treatment, CsA + UC-MSC significantly downregulated the histone methylation pathway (p < 0.05), whereas CsA + UCB-Treg significantly upregulated energy metabolism processes (p < 0.05). Treatment with CsA + UC-MSC upregulated superoxide dismutase activity compared with CsA + UCB-Treg treatment. Adding UC-MSC or UCB-Treg to CsA markedly enhanced the reconstruction of hematopoiesis in AA mice, with UC-MSC eliciting greater efficiency than UCB-Treg. Accordingly, the addition of these cells could further improve immune abnormalities.

Cyclosporine A alleviates colitis by inhibiting the formation of neutrophil extracellular traps via the regulating pentose phosphate pathway

BackgroundThe aberrant formation of neutrophil extracellular traps (NETs) has been implicated in ulcerative colitis (UC), a chronic recurrent intestinal inflammation. Cyclosporine A (CsA) is now applied as rescue therapy for acute severe UC. In addition, it has been certained that CsA inhibits the formation of NETs in vitro and the mechanism of which was still vague. The study aimed to explore the mechanism CsA inhibits the NETs formation of colitis in vivo and in vitro.MethodsNETs enrichment in clinical samples was analyzed using databases from Gene Expression Omnibus and verified in our center. Dextran sulfate sodium (DSS)-induced acute colitis mice model was used to investigate the effect of CsA on NETs of colonic tissue expression. To clarify the mechanism, intracellular energy metabolites were examined by Liquid Chromatograph Mass Spectrometer, and reactive oxygen species (ROS) levels were examined by fluorescence intensity in neutrophils treated with CsA after LPS stimulation. The transcriptional level and activity of G6PD of neutrophils were also assessed using qRT-PCR and WST-8. RNA Sequencing was used to detect differentially expressed genes of neutrophils stimulated by LPS with or without CsA. The expression levels of related proteins were detected by western blot.ResultsNETs enrichment was especially elevated in moderate-to-severe UC patients compared to HC. NETs expression in the colon from DSS colitis was decreased after CsA treatment. Compared with neutrophils stimulated by LPS, NETs formation and cellular ROS levels were decreased in LPS + CsA group. Cellular ribulose 5-phosphate and NADPH/NADP + related to the pentose phosphate pathway (PPP) were reduced in LPS + CsA group. In addition, CsA could decrease G6PD activity in neutrophils stimulated with LPS, and the results were further verified by inhibiting G6PD activity. At last, P53 protein was highly expressed in LPS + CsA group compared with the LPS group. Intracellular G6PD activity, ROS level and NETs formation, which were downregulated by CsA, could be reversed by a P53 inhibitor.ConclusionOur results indicated CsA could alleviate the severity of colitis by decreasing the formation of NETs in vivo. In vitro, CsA reduced ROS-dependent NETs release via downregulating PPP and cellular ROS levels by decreasing G6PD activity directly by activating the P53 protein.

Therapeutic Effects of Lycopene on Lipopolysaccharide-Induced Acute Lung Injury: Regulation of the PINK1/Parkin Signaling Pathway and Mitochondrial Autophagy

This study investigates the mechanism underlying the therapeutic effects of lycopene (Lyc) on lipopolysaccharide (LPS)-induced acute lung injury (ALI). ALI cell models were established using A549 cells treated with LPS. Cell viability was assessed using cell counting kit-8. ALI cells were treated with Lyc, mitochondrial autophagy inhibitor cyclosporine A (CsA), PTEN-induced kinase 1 (PINK1) activator Valinomycin, and reactive oxygen species (ROS) inhibitor N-acetylcysteine (NAC). Apoptosis was evaluated by flow cytometry and JC-1 probe staining. ROS and adenosine triphosphate levels were determined using dichloro-dihydro-fluorescein diacetate staining. Western blot assessed the expression levels of light chain 3 (LC3), Lysosomal-associated membrane protein 1, PINK1, and Parkin. An LPS-induced ALI rat model was treated with different concentrations of Lyc. Lung injury was assessed by hematoxylin and eosin staining. Levels of tumor necrosis factor-α, interleukin-6, and lactate dehydrogenase were measured by ELISA combined with magnetic nanoparticles. In this study, different concentrations of Lyc treatment enhanced cell survival. Additionally, Lyc treatment reduced the cell apoptosis rate, decreased lactate dehydrogenase (LDH) leakage and ROS level in ALI cells, and inhibited the expression of LC3, Beclin-1, PINK1, and Parkin. Consistent effects were seen in Lyc, CsA, Valinomycin, and NAC groups, suggesting similar impacts. In animal experiments, Lyc treatment significantly mitigated edema, inflammation, and autophagy in lung tissues. Furthermore, Lyc exhibited a protective effect on ALI cells by regulating PINK1/Parkin pathway and inhibiting mitochondrial autophagy. Overall, Lyc regulates mitochondrial autophagy in LPS-induced ALI with practical significance for studying its pharmacological mechanism and theoretical implications for understanding autophagy in ALI.

Adding Hetrombopag to Cyclosporine a to Patients with Newly Diagnosed Transfusion-Dependent Non-Severe Aplastic Anemia-a Prospective Phase 2 Study

Abstract Objective: To investigate the efficacy and safety of adding hetrombopag to cyclosporin A (CsA) in patients with transfusion-dependent non-severe aplastic anemia (TD-NSAA). Methods: This was a single center, prospective phase 2 study. Eligible patients were aged 18 years or older, were diagnosed as TD-NSAA within 6 months and treatment-naïve. Enrolled patients were treated with hetrombopag and CsA. Hetrombopag was started at 10mg/day with possible titration up to 15mg/day, and tapered gradually when the optimal response was achieved. CsA was given at 3-5mg/kg/d for at least 6 months, and tapered after the optimal response was reached. The primary endpoint was the overall response rate (ORR) and safety at 6 months. Patients who had been followed-up for at least 6 months were included in the analysis. A history cohort with compatible baseline characters treated with CsA alone were analyzed as control. The trial was registered with ClinicalTrails.gov NCT05018936. Result: Between Oct 2021 to Oct 2022, 28 patients were enrolled, comprising 13 men and 15 women, median age 46(range: 18-81). The ORR was 68.2% at 6 months. The most frequently reported adverse events were gastrointestinal discomfort grade 1 or 2. No grade 3 or 4 treatment-emergent adverse events were reported. There were 30 patients enrolled in the history control cohort with compatible baseline characters including age, sex ratio, complete blood cell count (P&amp;gt;0.05). After a median follow-up of 15 (range: 9-22) vs 16 (range:9-24) months(P&amp;gt;0.05), the ORR at 3, 6 months and the end of follow-up were 57.1% vs 16.7% (P=0.001), 68.2% vs 30.0% (P=0.006) and 90.9% vs 46.6%(P&amp;lt;0.001) in hetrombopag plus CsA group and CsA alone group, respectively. The complete response rate (CRR) at 3, 6 months and the end of follow-up were 10.7% vs 0% (P=0.068), 22.7% vs 10.0% (P=0.217) and 36.4% vs 16.6% (P=0.109) in both groups, respectively. Adverse effects were reported in 13 (46.4%) patients in hetrombopag plus CsA group and 15 (50%) patients of CsA alone group (P&amp;gt;0.05). 1 (3.6%) vs 8 (26.6%) patient relapsed (P=0.038) at 6 month in both groups. Clone evolution were noticed in no patients in hetrombopag plus CsA group and 2 (6.7%) patients in CsA alone group at 12 and15 months, respectively (P=0.225). Conclusion: Adding hetrombopag to CsA can improve the ORR for newly diagnosed TD-NSAA, with good tolerance and lower relapse rate. Keywords: Transfusion-dependent aplastic anemia, Hetrombopag, Cyclosporin A, efficacy, relapse.

Abstract 16801: Effect of Cyclosporine A on Infarct Size in Rat Hearts Donated After Circulatory Death

Introduction: Ischemia-reperfusion injury (IRI) is inherent in heart donation after circulatory death (DCD), hence only hearts with &lt;30 minutes of ischemia are used for transplantation. Mitochondrial permeability-transition pore (MPTP) opening has been implicated in IRI. Cyclosporine A (CyA) is known to inhibit MPTP opening. No data on infarct size reduction with CyA exists in a DCD heart. Hypothesis: CyA given during reperfusion will reduce infarct size in DCD hearts. Methods: Male Sprague-Dawley rats were assigned to either DCD + CyA, DCD + vehicle, or control-beating heart donors (CBD) (n=10 per group). Rats were anesthetized with pentobarbital, intubated and ventilated. Vecuronium and heparin were given for muscle paralysis and anticoagulation respectively. For DCD group, ventilation was terminated (start of anoxia/ischemia time), and hearts procured after 5, 10, 15, 20, 25 minutes. CBD hearts were procured without ischemia. Hearts were then perfused on a Langendorff setup for 90 minutes. Vehicle and CyA groups received DMSO and CyA (2μM) in KH buffer, respectively, for first 15 minutes followed by 75 minutes of KH buffer. Developed pressure (DP) was measured via a balloon tip catheter placed in the left ventricle. Infarct size was measured using triphenyltetrazolium chloride staining. CBD hearts perfused with KH buffer only served as controls. Results: Percent infarct size following 5, 10, 15, 20, and 25 minutes of ischemia was 17.7 ± 4, 19.3 ± 6, 25 ± 4, 27.3 ± 3, and 25.4 ± 11 respectively. CyA given to 25 minutes ischemia group reduced infarct size to 14.7%, corresponding to 6 minutes of ischemia. At 60 minutes CyA treated hearts had higher DP (mmHg) compared to vehicle group (34 ± 13 vs. 51 ± 17, p=0.017 ). Conclusions: CyA given during reperfusion to DCD hearts significantly reduced infarct size and preserved function. CyA use may allow for longer ischemia times during DCD heart procurement.

Anti-CD20 Antibody and Calcineurin Inhibitor Combination Therapy Effectively Suppresses Antibody-Mediated Rejection in Murine Orthotopic Lung Transplantation.

Antibody-mediated rejection (AMR) is a risk factor for chronic lung allograft dysfunction, which impedes long-term survival after lung transplantation. There are no reports evaluating the efficacy of the single use of anti-CD20 antibodies (aCD20s) in addition to calcineurin inhibitors in preventing AMR. Thus, this study aimed to evaluate the efficacy of aCD20 treatment in a murine orthotopic lung transplantation model. Murine left lung transplantation was performed using a major alloantigen strain mismatch model (BALBc (H-2d) → C57BL/6 (BL/6) (H-2b)). There were four groups: isograft (BL/6→BL/6) (Iso control), no-medication (Allo control), cyclosporine A (CyA) treated, and CyA plus murine aCD20 (CyA+aCD20) treated groups. Severe neutrophil capillaritis, arteritis, and positive lung C4d staining were observed in the allograft model and CyA-only-treated groups. These findings were significantly improved in the CyA+aCD20 group compared with those in the Allo control and CyA groups. The B cell population in the spleen, lymph node, and graft lung as well as the levels of serum donor-specific IgM and interferon γ were significantly lower in the CyA+aCD20 group than in the CyA group. Calcineurin inhibitor-mediated immunosuppression combined with aCD20 therapy effectively suppressed AMR in lung transplantation by reducing donor-specific antibodies and complement activation.

No Difference Between Tacrolimus and Cyclosporine A on Depression Among Kidney Transplantation Recipients

The 2 main types of calcineurin inhibitors (CNI) are tacrolimus (TAC) and cyclosporine A (CsA); both are needed by patients who receive kidney transplants. A common adverse reaction of TAC is depression, which is listed in its instructions. However, depression occurred rarely, according to the instructions manual for CsA. Scales measuring depression were sent to recipients who had taken TAC or CsA to observe whether there was a difference in depression between patients who consumed the 2 drugs. From September 23rd-December 8th 2022, a questionnaire was sent to kidney transplant recipients online to investigate depression by PHQ-9 score. Then, the questionnaires returned were divided into 2 groups: TAC group and CsA group. The difference of basic characteristics was made to equal by means of propensity score matching (PSM). The scores, degrees of depression, and prevalence of major depression between the 2 groups were compared. Of 259 questionnaires returned, 220 questionnaires were valid. Among them, 170 recipients used TAC and 50 recipients used CsA. There were no significant differences in baseline characteristics after PSM. After PSM, there was no statistically significant difference in PHQ-9 (0.8) score, degree of depression (P=.7), or rate of major depression between the 2 groups. There was no significant difference between kidney transplant recipients taking TAC or CsA in PHQ-9 score, degree of depression, or prevalence of major depression.

Minocycline Prevents Depression-like Behavior After Co-administration With Dexamethasone or Cyclosporine-A in Mice

Background: In animal studies, minocycline (Mcy) has been proven to have antidepressant effects. In addition to modulating peripheral and central pro-inflammatory pathways, Mcy may regulate the hypothalamic-pituitary-adrenal (HPA) axis and the mechanistic target of rapamycin (mTOR) signaling pathway. This study aims to evaluate the antidepressant-like effect of Mcy in mice following injection of dexamethasone (Dex) or cyclosporine-A (CsA). Methods: Male NMRI mice were randomly divided into eight groups of 6, including control, Dex 0.25 mg/kg, CsA 20 mg/kg, Mcy 40 mg/kg, Dex+Mcy, Dex+fluoxetine 20 mg/kg, CsA+Mcy, and CsA+fluoxetine. All drugs were injected intraperitoneally (except for Dex, which was subcutaneous injection) once daily for 3 days. The locomotor activity, forced swimming test (FST), and sucrose preference (SP) test were performed on day 4. Results: Mcy alone reduced immobility time in the FST (27.0±6.4 s) compared to the control group (104±3.9 s) (P&lt;0.001). After the co-administration of Mcy and Dex, the immobility time significantly decreased (79.5±6.5 s) compared to the Dex group (P&lt;0.001). It also decreased following the co-administration of Mcy and CsA (67.5±20.8 s) compared to the CsA group (P&lt;0.001). Results were similar in the groups treated with fluoxetine plus Dex or CsA. Significant differences were not observed in the locomotor activity test. Conclusion: Mcy prevents depression-like behavior in mice during the FST when it is co-administrated with CsA or Dex. The possibility of the positive effect of Mcy on the HPA axis and the mTOR signaling pathway should be examined in further studies.

Cyclosporine A induces Epstein–Barr virus reactivation in tree shrew (Tupaia belangeri chinensis) model

Epstein–Barr virus (EBV) usually exists as a latent infection in immunocompetent hosts but immunosuppressed individuals are at risk for developing EBV reactivation that leads to the uncontrolled proliferation of B lymphocytes. In this study, we have mimicked the immunosuppressed microenvironment in the tree shrew model of EBV infection by using cyclosporine A (CsA). The results showed that EBV-cocultured peripheral blood mononuclear cells (PBMCs) proliferated vigorously in response to CsA treatment in vitro. However, EBV susceptibility in vivo depended on the timing of CsA administration. Reactivation of EBV occurred in the latently EBV-infected tree shrews after treatment with 25 mg/kg/day CsA (EBV > CsA group), whereas tree shrews were no longer susceptible to infection if CsA was administered for five weeks before EBV injection (CsA > EBV group). RNA-seq analysis of both groups identified a further link between immunosuppression and EBV infection. KEGG pathway enrichment analysis revealed a significant enrichment of viral infection-related pathways in the EBV > CsA group, whereas tumor-related pathways were significantly enriched in the CsA > EBV group. A protein–protein interaction network was constructed using Cytoscape for the purpose of identifying hub genes that were then verified using qRT-PCR. In conclusion, the tree shrew model of EBV infection exhibits certain features of EBV infection in humans and serves as a valuable platform for exploring the underlying mechanisms of EBV infection.

Comparison of Cyclosporine A and Cyclosporine A Combined with Corticosteroid in the Treatment of Acquired Pure Red Cell Aplasia

To evaluate the efficacy, safety and relapse of cyclosporine A (CsA) and CsA combined with corticosteroid (CS) as the frontline therapy for patients with newly diagnosed acquired pure red cell aplasia (aPRCA). The clinical features, treatment responses, relapses and clinical outcomes of patients with newly diagnosed aPRCA in Peking Union Medical College Hospital (PUMCH) from January 2015 to May 2020 were analyzed retrospectively. All the enrolled patients had been treated with either CsA or CsA+CS for at least 6 months and had been followed up for at least 12 months, with complete clinical data and consent forms. 96 patients including 72 treated with CsA and 24 treated with CsA+CS were enrolled. With comparable baseline characteristics and follow-up periods, patients treated with CsA or with CsA+CS had similar overall response rates (ORRs) and complete response rates (CRRs) at the 3rd, 6th and 12th month and at the end of follow-up (P>0.05). Meanwhile, no significant difference was found between the two groups in the optimal ORR, optimal CRR, time to response or time to complete response. CsA+CS and CsA groups had similar adverse event (AE) rates, but CsA+CS group had higher CS-related infection rate (P <0.05). One patient in CsA+CS group died of multiple infections. As for the relapse, the two groups had compatible relapse rates at different time points, time to relapse, overall relapse rate and relapse-free survival (P>0.05). CsA exposure time, rather than different therapy regimens, was the only influence factor for either ORR or relapse rate (P <0.05). CsA monotherapy has similar efficacy, AE rate and relapse rate as compared with CsA+CS for patients with newly diagnosed aPRCA, and shows less CS-related AEs such as infection.

Omega-3 fatty acids upregulate Nrf2 expression and attenuate apoptosis, inflammation, and fibrosis in a rat model of cyclosporine-induced nephropathy

Background: Cyclosporine A (CsA)-induced kidney injury is characterized by renal impairment with inflammatory cell infiltrations, apoptosis, fibrosis, and hypoxic injury. It is not clear whether omega-3 fatty acids (O-3 FAs), which have anti-inflammatory and antioxidant roles, affect nuclear factor erythroid 2-related factor 2 (Nrf2) expression. The aim of this study was to investigate whether O-3 FAs affect Nrf2 expression and exert anti-inflammatory, anti-apoptotic, and anti-fibrotic effects in CsA-induced nephropathy.Methods: Male Sprague-Dawley rats were divided into control, CsA-treated, and CsA-treated with O-3 FA groups. Nrf2 expression was measured by Western blots and immunohistochemical staining.Results: Kidney function was impaired in the CsA-treated rats compared to the controls. Caspase-3 and caspase-7 were activated in the CsA-treated group, and the Bax/Bcl2 ratio was higher. O-3 FAs attenuated these apoptosis-related changes. ED-1 and inhibition of kappa B (IĸB) protein expression were significantly upregulated in the CsA-treated group. Compared to the control group, O-3 FA supplementation attenuated the increased expression of ED-1 and IĸB related to inflammation. Smad2/3, Smad4, and transforming growth factor-β1 were activated in the CsA group, and O-3 FA treatment prevented these changes related to renal fibrosis. The expression of Nrf2 was reduced in CsA-treated rats, but Nrf-2 was increased by O-3 FA treatment.Conclusions: We suggest that Nrf2 is a potential mediator induced by O-3 FA supplementation and that it attenuates pro-inflammatory pathways, fibrotic processes, and apoptosis. Further studies are needed to elucidate the crosstalk between Nrf2 expression and signals related to O-3 FA treatment.

Cyclosporine A induces cardiac remodeling through TGF-β/Smad3/miR-29 signaling pathway and alters gene expression of miR-30b-5p/CaMKIIδ isoforms pathways: alleviating effects of moderate exercise.

Cyclosporine A (CsA)-induced cardiac interstitial fibrosis and cardiac hypertrophy are highly known phenomena; however, the basic mechanisms of CsA cardiotoxicity are unclear. The present study evaluated the role of the Transforming growth factor-beta (TGF-β)/Smad3/miR-29b signaling pathway and CaMKIIδ isoforms gene expression in cardiac remodeling under CsA exposure alone or combined with moderate exercise. A total of 24 male Wistar rats were divided into control, cyclosporine (30mg/kg BW), and cyclosporine-exercise groups. After 42 days of treatment, the findings revealed a significant decline in miR-29 and miR-30b-5p gene expression and an increase in gene expression of Smad3, calcium/calmodulin-dependent protein kinaseIIδ (CaMKIIδ) isoforms, Matrix Metalloproteinases (MMPs), protein expression of TGF-β, heart tissue protein carbonyl and oxidized LDL (Ox-LDL), and plasma LDL and cholesterol levels in the CsA-treated group compared to the control group. The CsA group presented greater histological heart changes such as fibrosis, necrosis, hemorrhage, infiltrated leukocyte, and left ventricular weight/heart weight than the control group. Moreover, combined moderate exercise and CsA relatively improved gene expression changes and histological alternations compared to the CsA group. TGF-β-Smad3-miR-29 and CaMKIIδ isoforms may mainly contribute to the progression of heart fibrosis and hypertrophy due to CsA exposure, providing new insight into the pathogenesis and treatment of CsA-induced side effects on the heart tissue.

Comparison of topical application of cyclosporine A to prednisolone in treatment of dry eye disease

Background Dry eye disease (DED) is a common ophthalmic disorder which affects 5–50% of the global population. Aim The aim of this study was to compare the effects of topical application of cyclosporine A (CsA) to prednisolone in patients with DED. Patients and methods The study included 40 patients diagnosed with DED. They were randomly divided into two groups: group A patients received treatment with topical 0.05% CsA twice a day, and group B patients received treatment with topical 1% prednisolone four times a day for 1 month. All patients were examined at three visits: before treatment, after 2 weeks, and 1 month after the intervention to show the effect of the two drugs on the tear break-up time (TBUT), Schirmer’s score, ocular surface disease index (OSDI), and surface stain, in addition to assessment of the patient’s DED-related symptoms. Results In the prednisolone group, the pretreatment Schirmer’s test results were 3.05±1.57 and 2.35±1.57 mm for the right (OD) and left eyes (OS), respectively. It improved to 5.65±2.30 and 5.85±2.43 mm, respectively 2 weeks after discontinuation of treatment (posttreatment). Similarly, the pretreatment values for the CsA group were 1.35±1.57 and 2.0±1.52 mm and became 5.65±2.30 and 5.85±2.434 mm, respectively after treatment. There was a statistically significant difference between the two groups regarding the pretreatment (P=0.001) and posttreatment (P=0.026) Schirmer’s test results. In the prednisolone group, the pretreatment TBUT was 4.20±1.61 and 3.85±1.09 s and improved to 6.60±1.31 and 6.85±1.42 s, respectively after treatment. Similarly, in the CsA group the pretreatment values were 2.65±1.81 and 3.30±1.38 s and became 8.65±1.69 and 9.30±1.42 s, respectively, posttreatment. In addition, there was a statistically significant difference between the studied groups regarding the pretreatment (P=0.007) and posttreatment TBUT (P&lt;0.001). The pretreatment OSDI, in the prednisolone group, was 47.28±5.31 and became 38.00±4.33 posttreatment. Similarly, in the CsA group, the pretreatment value was 49.18±8.69 and became 35.03±8.04 posttreatment. However, there were no statistically significant differences between the studied groups regarding the pretreatment and posttreatment OSDI (P&gt;0.05). The surface stain improved in both groups after treatment, with the prednisolone group showing the greatest improvement at 2 weeks (P=0.002). However, there was no statistically significant difference between the two groups at 1 month (P&gt;0.05). Conclusion Both topical CsA and prednisolone demonstrated effectiveness in treating moderate to severe DED cases and were comparable. Prednisolone showed a greater improvement in OSDI (that assesses the DED-related symptoms) and surface stain, while CsA was associated with more improvement in Schirmer’s test and TBUT.

Niclosamide modulates cyclosporin A-induced hepatotoxicity in a mouse model: PPAR-γ and Wnt/β-catenin crosstalk.

The aim of this study was to evaluate whether: 1) Wnt/β-catenin signaling is involved in cyclosporin A (CsA)-induced hepatotoxicity, and 2) knockdown of this pathway by niclosamide (NCL) attenuate CsA-induced hepatotoxicity. The experiment was accomplished in 21days. Adult male mice were randomly distributed into five groups: control group, CsA (25mg/kg/day) group, CsA+NCL (2.5mg/kg/day) group, CsA+NCL (5mg/kg/day) group, and NCL (5mg/kg/day) group. NCL showed marked hepatoprotection by significantly decreasing liver enzymes activities and ameliorating the histopathological alterations induced by CsA. Besides, NCL alleviated oxidative stress and inflammation. NCL-treated groups (2.5 and 5mg/kg) displayed rise in the expression of hepatic peroxisome proliferator-activated receptor-γ (PPAR-γ) by 2.1- and 2.5-fold, respectively. Notably, NCL (2.5 and 5mg/kg) significantly inhibited Wnt/β-catenin signaling, evidenced by a marked decrease in the hepatic expression of Wnt3a by 54% and 50%, frizzled-7 receptor by 50% and 50%, β-catenin by 22% and 49%, and c-myc by 50% and 50%, respectively. NCL can be regarded as a potential agent to mitigate CsA-induced hepatotoxicity.

Short‐term cyclosporin A treatment improves pregnancy outcomes in patients with unexplained pregnancy loss: A prospective, open‐label, observational study

Short‐term cyclosporin A treatment improves pregnancy outcomes in patients with unexplained pregnancy loss: A prospective, open‐label, observational study

Comparative efficacy of mycophenolic acid and cyclosporine A in treatment of children with steroid-dependent nephrotic syndrome

BACKGROUND. Steroid-dependent nephrotic syndrome (SDNS) treatment is still an important problem in pediatric nephrology since the proper use of steroid-sparing agents can reduce the frequency of relapses and avoid steroid toxicity. THE AIM of our study was to compare the efficacy of cyclosporine A (CsA) and mycophenolic acid (MPA) in children with SDNS. PATIENTS AND METHODS. We observed 91 children (30 girls, 61 boys) with SDNS in 2015-2020. Age at the disease debut was 3 years [2,1; 5,0]. The age at steroid-sparing therapy debut was 8 years [4,0; 16,0]. All children received standard clinical and laboratory examinations and kidney biopsy. RESULTS: 25 children were treated with CsA, and 66 children were treated with MPA. The steroid-sparing therapy duration was 36 months [29; 44]. The frequency of relapses before the steroid-sparing therapy onset was 1,32 ± 0,62 (0,5; 4,3)/year, during steroid-sparing therapy it became 0,5 ± 0,58 (0; 2)/year (р &lt; 0,05). Relapse rate in the MPA group was 0,36 ± 0,49 (0; 1,76)/year compared to 0,85 ± 0,66 (0; 2)/year in the CsA group (p &lt; 0,05). Withdrawal of prednisolone was achieved in 44 (48,4 %) children. The relapse-free period during steroid-sparing therapy with steroid withdrawal was 25 months [6; 120]. 15 children (16,5 %) showed long–term stable remission with no immunosuppressive therapy. 14 of them were treated with MPA and only one with CsA (χ2=9,7, р = 0,0021). The remission duration was 7-32,9 months. There were no severe side effects of CsA and MPA requiring discontinuation of therapy. CONCLUSION: steroid-sparing therapy of SDNS with CsA and MPA significantly reduces relapse frequency. Relapse risk in patients treated with CsA was significantly higher than with MPA. So, it is justified to prescribe MPA as a first-choice immunosuppressive therapy in patients with SDNS.

Immunomodulatory effect of pachymaran on cyclosporine A (CsA)-induced lung injury in mice

ObjectiveTo investigate the immunomodulatory effect of pachymaran on cyclosporine A (CsA)-induced lung injury in mice. Methods(i) Fifty male BALB/c mice were randomly divided into five groups (10 mice in each group): normal control (NC) group, 30, 45, and 60 mg/kg CsA groups, and lipopolysaccharide (LPS) group. Except for the NC group, other groups underwent CsA modeling. The NC group was treated with phosphate-buffered saline (PBS), the LPS group with 10 mg/kg LPS eight hours before mice euthanized, and the 30, 45, and 60 mg/kg CsA groups with corresponding doses of CsA for seven consecutive days. After treatment, the body and organ mass of each group were weighed, and the lung, thymus, and spleen indexes were calculated. Hematoxylin-Eosin (HE) staining was performed to observe histopathological changes in the lungs of the mice. The protein expression levels of interleukin (IL)-2 and IL-1β in the blood were detected using enzyme-linked immunosorbent assay (ELISA), and those of surfactant protein D (SP-D), IL-2, and IL-6 in lung tissues were detected by immunohistochemistry (IHC). The mRNA expression levels of SP-D, IL-1β, IL-6, and myeloperoxidase (MPO) in the lung tissues were detected by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). (ii) Another 60 BALB/c mice were divided into six groups (10 mice in each group) : NC group, model control (MC) group, 50, 100, and 200 mg/kg pachymaran groups, and polyinosinic-polycytidylic acid [poly(I:C)] group. Except for the NC group, other groups underwent 45 mg/kg CsA modeling. The NC and MC groups were treated with distilled water, the pachymaran groups with corresponding doses pachymaran, and the poly(I:C) group with 0.1 mg/kg poly(I:C) for seven days.The mice were euthanized to obtain tissues and serum for detection. Detection methods were identical to those described in (i) above. Results(i) CsA (30 mg/kg) increased the lung index of mice (P < 0.001), and decreased the spleen index (P < 0.01), thymus index (P < 0.05), and the serum level of IL-2 (P < 0.05). CsA (45 mg/kg) decreased the spleen, thymus indexes, and the serum level of IL-2 (P < 0.01) in mice, and increased the serum level of IL-1β (P < 0.05) and the protein level of lung SP-D (P <0.001). CsA (60 mg/kg) increased the lung index of mice (P < 0.01), the serum level of IL-1β (P < 0.05), the protein level of lung SP-D (P < 0.01), and the mRNA levels of lung MPO and SP-D ( P < 0.05), and decreased the thymus index of mice (P < 0.01). HE staining showed that 30, 45, and 60 mg/kg CsA, and LPS caused pathological changes in the lung tissue of mice. (ii) After pachymaran intervention in MC mice, the spleen and thymus indexes (P < 0.05) were increased in the 100 and 200 mg/kg pachymaran groups, and the lung index was decreased (P < 0.05). Moreover, 50 mg/kg pachymaran increased the thymus index (P < 0.05) and decreased the lung index (P < 0.01) in MC group. Pachymaran (50, 100, and 200 mg/kg) improved lung tissue injury, reduced the serum level of IL-1β (P < 0.001), and the mRNA levels of MPO and SP-D in lung tissues (P < 0.05) of mice. Pachymaran (100 mg/kg) increased the protein level of lung IL-2 (P < 0.01), decreased the protein level of lung SP-D (P < 0.01), and the mRNA level of IL-1β (P < 0.001) in the lung tissues of mice. Pachymaran (200 mg/kg) increased the serum level of IL-2 (P < 0.01) and lung IL-6 of mice (P < 0.05). Pachymaran (50 and 200 mg/kg) increased the mRNA level of IL-6 in the lung tissues of mice (P < 0.05). ConclusionWhile the immune function of mice was suppressed by CsA, the lung tissue was also damaged. Pachymaran can improve the immunosuppression induced by CsA and improve the lung tissue injury in immunosuppressed mice.