Effects of Yinzhihuang on Alleviating Cyclosporine A-Induced Cholestatic Liver Injury via Farnesoid X Receptor-Mediated Regulation of Transporters and Enzymes in Vitro and in Vivo.

Yinzhihuang (YZH), a traditional Chinese medicine prescription, was widely used to treat cholestasis. Cholestatic liver injury limited the use of the immunosuppressive drug cyclosporine A (CsA) in preventing organ rejection after solid organ transplantation. Clinical evidences suggested that YZH could enhance bile acids and bilirubin clearance, providing a potential therapeutic strategy against CsA-induced cholestasis. Nevertheless, it remains unclear whether YZH can effectively alleviate CsA-induced cholestatic liver injury, as well as the molecular mechanisms responsible for its hepatoprotective effects. The purpose of the present study was to investigate the hepatoprotective effects of YZH on CsA-induced cholestatic liver injury and explore its molecular mechanisms in vivo and vitro. The results demonstrated that YZH significantly improved the CsA-induced cholestatic liver injury and reduced the level of liver function markers in serum of Sprague-Dawley (SD) rats. Targeted protein and gene analysis indicated that YZH increased bile acids and bilirubin efflux into bile through the regulation of multidrug resistance-associated protein 2 (Mrp2), bile salt export pump (Bsep), sodium taurocholate cotransporting polypeptide (Ntcp) and organic anion transporting polypeptide 2 (Oatp2) transport systems, as well as upstream nuclear receptors farnesoid X receptor (Fxr). Moreover, YZH modulated enzymes involved in bile acids synthesis and bilirubin metabolism including Cyp family 7 subfamily A member 1 (Cyp7a1) and uridine 5'-diphosphate (UDP) glucuronosyltransferase family 1 member A1 (Ugt1a1). Furthermore, the active components geniposidic acid, baicalin and chlorogenic acid exerted regulated metabolic enzymes and transporters in LO2 cells. In conclusion, YZH may prevent CsA-induced cholestasis by regulating the transport systems, metabolic enzymes, and upstream nuclear receptors Fxr to restore bile acid and bilirubin homeostasis. These findings highlight the potential of YZH as a therapeutic intervention for CsA-induced cholestasis and open avenues for further research into its clinical applications.

Effects of Cyclosporine A and Adalimumab on the expression profiles histaminergic system-associated genes and microRNAs regulating these genes in HaCaT cells.

Previous studies have not completely elucidated the role of the histaminergic system in the pathogenesis of psoriasis. This study aimed to evaluate the effects of adalimumab and cyclosporine A on the expression of histaminergic system-related genes and miRNAs regulating these genes in bacterial lipopolysaccharide A (LPS)-stimulated human keratinocyte (HaCaT) cells. HaCaT cells were treated with 1 µg/mL LPS for 8 h, followed by treatment with 8 µg/mL adalimumab or 100 ng/mL cyclosporine A for 2, 8, or 24 h. Untreated cells served as controls. The cells were subjected to ribonucleic acid (RNA) extraction and microarray, quantitative real-time polymerase chain reaction, and enzyme-linked immunosorbent assay analyses. Statistical analysis was performed using the Statistica 13.0 PL (StatSoft, Cracow, Poland) and the Transcriptome Analysis Console programs (Affymetrix, Santa Clara, CA, USA) (p < 0.05). The differential expression of the following two miRNAs was not affected in LPS-stimulated cells upon treatment with cyclosporine A or adalimumab: hsa-miR-583 (downregulated expression), involved in the regulation of histamine receptor 1 - HRH1 (overexpression); has-miR-1275 (downregulated expression), involved in the regulation of histamine receptor 1 - HRH3 (overexpression) and Solute carrier family 22 member 3 - SLC23A2 (downregulated expression)). Adalimumab and cyclosporine A modulated the histaminergic system in HaCaT cells in vitro. However, further studies are needed to elucidate the underlying mechanisms.Abbreviations: (-) - downregulated in comparison to the control, (+) - overexpression in comparison to the control, ACTB - ß-actine, ADA - Adenosine deaminase, ADCYAP1 - Adenylate Cyclase Activating Polypeptide 1, BMP - bone morphogenetic protein, bp - base pair, cAMP - adenosine 3' 5'-cyclic monophosphate, CBX7 - Chromobox protein homolog 7, cDNA - double-stranded complementary DNA, CSA - cyclosporine A DAG - diacylglycerol, DIAPH - Diaphanous related formin 1, DNMT - DNA methyltransferases, DRD2 - Dopamine receptor D2, EDN1 - Endothelin 1, EDNRA - Endothelin receptor type A, ELISA - Enzyme-linked immunosorbent assay, EZH2 - Enhancer of zeste homolog 2, FC - fold change, GABRB1 - Gamma-aminobutyric acid (GABA) A receptor, alpha 1, GABRB2 - Gamma-aminobutyric acid (GABA) A receptor, alpha 2, GABRB3 - Gamma-aminobutyric acid (GABA) A receptor, alpha 3, HaCaT - Human adult, low-calcium, high-temperature keratinocytes, HIS - Human Histamine, HLAs - human leukocyte antigens, HNMT - Histamine N-methyltransferase, HNMT - Histamine N-Methyltransferase, HRH1 - histamine receptor 1, HRH2 - histamine receptor 2, HRH3 - histamine receptor 3, HRH4 - histamine receptor 4, HTR6 - 5-Hydroxytryptamine Receptor 6, IGF1 - Insulin-like growth factor 1, IL10 -interleukin 10, IL12 -interleukin 12, IL6 - interleukin 6, IP3 - inositol 1,4,5-triphosphate, LPS - bacterial lipopolysaccharide A, LYN - LYN Proto-Oncogene, Src Family Tyrosine Kinase, MAPKs -mitogen-activated protein kinases, miRNA - micro RNA, MMP2 - matrix metalloproteinase-2, NHDF - Normal Human Dermal Fibroblasts, NHEK - Normal Human Epidermal Keratinocytes, OCT3 - organic cation transporter 3, PANTHER - Protein ANalysis THrough Evolutionary Relationships Classification, PBS - phosphate-buffered saline, PI3K-AKT - phosphatidylinositol 3-kinase-protein kinase B, PIP2 - phosphatidylinositol 4,5 bisphosphate, PMSF - phenylmethylsulfonyl fluoride, PSORS1- psoriasis susceptibility gene 1, qRT-PCR - quantitative Reverse Transcription Polymerase Chain Reaction, RNA - ribonucleic acid, RNAi - RNA interference, RTqPCR - Real-Time Quantitative Reverse Transcription Reaction, SLC223A2 - Solute carrier family 22 member 3, SNX -Sorting nexin, SOX9 - SRY-Box Transcription Factor 9, TGF-α - transforming growth factor α, TGF-ß - transforming growth factor beta, TNF-α - tumor necrosis factor alpha, TP53 - tumor protein 5 z, VAMP2 - Vesicle associated membrane protein 2.

Erythrodermic psoriasis in post-coronavirus disease 2019 patient.

Erythrodermic psoriasis (EP) is characterized by generalized erythema and desquamation affecting more than 75% of body surface area and usually accompanied by systemic symptoms. The triggers are medication withdrawal, drugs reactions, and systemic infections including coronavirus disease 2019 (COVID-19). A 46-year-old man with plaque psoriasis suffered from EP following the sudden discontinuation of medications. He was diagnosed with COVID-19 one month before erythroderma appeared. The body surface area involvement was 96% and psoriasis area severity index was 49.8. His general condition and laboratory examination were within normal limits. He was treated with cyclosporine-A for one month after being healed from COVID-19 with significant improvement. Excessive production of proinflammatory cytokines in COVID-19 plays a role in the pathogenesis of psoriasis. This condition should be managed appropriately to minimize the complication. Cyclosporine-A is the first-line therapy for EP because of its effectiveness and good safety profile. It is also shown a beneficial effect in COVID-19 infection in vitro.

Effect of Wuzhi capsules on cyclosporine A concentration in children with aplastic anemia immunotherapy: a single-center observational study.

OBJECTIVE: This research aimed to assess the effect of Wuzhi capsules (WZC) on the blood concentration of cyclosporine A (CsA) in renal aplastic anemia recipients. METHODS: This observational study was carried out at the Hematology Oncology Center, Beijing Children's Hospital between November 2019 and February 2020. A total of 102 Chinese AA recipients receiving CsA (6 mg/kg/d) with or without WZC were included in this study. Baseline data, such as age, therapeutic drug monitoring data, and follow-up information were collected. The promotion concentration of CsA was calculated, and the pharmaceutical economics evaluation with combination of two drugs was also carried out. RESULTS: Dose- and body weight-adjusted trough concentrations (C0/D/W) of CsA in the WZC group were found to be significantly higher than that in the non-WZC group (P < 0.01). The average C0 of CsA increased by (63.27 ± 45.81) ng/mL. The incidence of adverse events was also not statistically significant between the two groups (P > 0.05). CONCLUSION: WZC can increase CsA concentration without increasing adverse drug reactions. Efficient and convenient immunosuppressive effects on AA recipients can be achieved via immunosuppressant therapy in combination with WZC.

The Protective Effect of Oral Application of Corni Fructus on the Disorders of the Cornea, Conjunctiva, Lacrimal Gland and Retina by Topical Particulate Matter 2.5.

Particulate matter 2.5 (PM2.5) may aggravate dry eye disease (DED). Corni Fructus (CF), which is fruit of Cornus officinalis Sieb. et Zucc., has been reported to have various beneficial pharmacological effects, whereas the effect of CF on the eye is still unknown. Therefore, in this study, we investigated the effect of oral administration of water extract of CF (CFW) on the eye, hematology, and biochemistry in a DED model induced by topical exposure to PM2.5. Furthermore, the efficacy of CFW compared with cyclosporine (CsA), an anti-inflammatory agent, and lutein, the posterior eye-protective agent. Sprague-Dawley rats were topically administered 5 mg/mL PM2.5 in both eyes four times daily for 14 days. During the same period, CFW (200 mg/kg and 400 mg/kg) and lutein (4.1 mg/kg) were orally administered once a day. All eyes of rats in the 0.05% cyclosporine A (CsA)-treated group were topically exposed to 20 µL of CsA, twice daily for 14 days. Oral administration of CFW attenuated the PM2.5-induced reduction of tear secretion and corneal epithelial damage. In addition, CFW protected against goblet cell loss in conjunctiva and overexpression of inflammatory factors in the lacrimal gland following topical exposure to PM2.5. Furthermore, CFW markedly prevented PM2.5-induced ganglion cell loss and recovered the thickness of inner plexiform layer. Meanwhile, CFW treatment decreased the levels of total cholesterol and low-density lipoprotein cholesterol in serum induced by PM2.5. Importantly, the efficacy of CFW was superior or similar to that of CsA and lutein. Taken together, oral administration of CFW may have protective effects against PM2.5-induced DED symptoms via stabilization of the tear film and suppression of inflammation. Furthermore, CFW may in part contribute to improving retinal function and lipid metabolism disorder.

In vitro effects of tropisetron and granisetron against Echinococcus granulosus (s.s.) protoscoleces by involvement of calcineurin and calmodulin.

BACKGROUND: Cystic echinococcosis (CE) is a disease caused by the larval stage of Echinococcus granulosus sensu lato  (s.l.). The treatment of CE mainly relies on the use of benzimidazoles, which can commonly cause adverse side effects. Therefore, more efficient treatment options are needed. Drug repurposing is a useful approach for advancing drug development. We have evaluated the in vitro protoscolicidal effects of tropisetron and granisetron in E. granulosus sensu stricto (s.s.) and assessed the expression of the calcineurin (CaN) and calmodulin (CaM) genes, both of which have been linked to cellular signaling activities and thus are potentially promising targets for the development of drugs. METHODS: Protoscoleces (PSC) of E. granulosus (s.s.) (genotype G1) obtained from sheep hepatic hydatid cysts were exposed to tropisetron and granisetron at concentrations of 50, 150 and 250 µM for various periods of time up to 10 days. Cyclosporine A (CsA) and albendazole sulfoxide were used for comparison. Changes in the morphology of PSC were investigated by light microscopy and scanning electron microscopy. Gene expression was assessed using real-time PCR at the mRNA level for E. granulosus calcineurin subunit A (Eg-CaN-A), calcineurin subunit B (Eg-CaN-B) and calmodulin (Eg-CaM) after a 24-h exposure at 50 and 250 µM, respectively. RESULTS: At 150 and 250 µM, tropisetron had the highest protoscolicidal effect, whereas CsA was most effective at 50 µM. Granisetron, however, was less effective than tropisetron at all three concentrations. Examination of morphological alterations revealed that the rate at which PSC were killed increased with increasing rate of PSC evagination, as observed in PSC exposed to tropisetron. Gene expression analysis revealed that tropisetron at 50 µM significantly upregulated Eg-CaN-B and Eg-CaM expression while at 250 µM it significantly downregulated both Eg-CaN-B and Eg-CaM expressions; in comparison, granisetron decreased the expression of all three genes at both concentrations. CONCLUSIONS: Tropisetron exhibited a higher efficacy than granisetron against E. granulosus (s.s.) PSC, which is probably due to the different mechanisms of action of the two drugs. The concentration-dependent effect of tropisetron on calcineurin gene expression might reflect its dual functions, which should stimulate future research into its mechanism of action and evaluation of its potential therapeutical effect in the treatment of CE.

Evaluation of different almond (Terminalia catappa) extracts against oxidative stress induced by cyclosporine in brain and liver of rats.

OBJECTIVES: This study was designed to evaluate the ameliorative effect of almond (Terminalia catappa) leaf (ALE) and stem bark (ABE) extracts on the enzyme activities and oxidative stress markers in the brain and liver tissues of cyclosporine-A (CsA) stressed male albino rats. METHODS: Eighty-eight adult male rats weighing between 200 and 220 g were randomly distributed to into 11 groups (n=8) and different doses (100 and 200 mg/kg bwt.) of ALE and ABE were administered through oral gavages to the normal rats and 50 mg/kg/bwt/day CsA-stressed, while normal control rats was given a saline solution (p.o), and the treatment lasted for 14 days. Blood plasma, liver and brain tissues were prepared for biochemical analysis. RESULTS: Neuronal [acetylcholinesterase (AChE) and butrylcholinesterase (BChE) and arginase] enzyme activities and thiobarbituric acid reactive species (TBARS) level, plasma aspartate transferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) activities, liver non-protein thiol (NPSH) level were analyzed. The results revealed that, the administration of CsA induced a significant increase in neuronal AChE, BChE, arginase, TBARS level, but decreased nitric oxide (NO) level. CsA also increased ALT, AST, and ALP activities in the blood plasma of CsA stress rats compared to normal control, but were significantly reversed respectively (p<0.001) upon treatment with the ALE and ABE dose-dependently. CONCLUSIONS: The study revealed that ALE and ABE could prevent neuronal dysfunction and liver toxicity induced by CsA administration, however, higher dose (200 mg/kg) of the studied extracts appears to be more potent.

Schisandrae chinensis fructus extract protects against hepatorenal toxicity and changes metabolic ions in cyclosporine A rats.

While cyclosporin A (CsA) is an effective immunosuppressive agent, its clinical application is limited by serious hepatorenal toxicity. However, Schisandrae chinensis fructus extract (SCE) has been previously shown to alleviate the hepatorenal damage caused by CsA. In this study, we aimed to evaluate the protective effects of SCE against hepatorenal toxicity induced by CsA. Our results revealed that SCE can prevent and treat CsA-induced liver and kidney injury. Furthermore, SCE inhibited the upward trend of dUDP and CDP-ethanolamine in the urine of CsA rats, pathways of which are involved in pyrimidine and glycerophospholipid metabolism. We finally confirmed that this protection of SCE was regulated by the activation of Nrf2 signaling pathway and the inhibition of apoptosis. In summary, our findings indicated that SCE may effectively prevent and treat hepatorenal toxicity caused by CsA. In addition, metabolomic techniques identified potential biomarkers for the occurrence of hepatorenal toxicity in CsA rats.

Inhibition effect of epigallocatechin-3-gallate on the pharmacokinetics of calcineurin inhibitors, tacrolimus, and cyclosporine A, in rats.

BACKGROUND: Epigallocatechin-3-gallate (EGCG) is the most biologically active catechin of green tea. Tacrolimus (TAC) and cyclosporine A (CsA) are immunosuppressive agents commonly used in clinical organ transplantation. The present study investigated the effect of EGCG on the pharmacokinetics of TAC and CsA in rats and its underlying mechanisms. RESEARCH DESIGN AND METHODS: Either TAC or CsA was administered to rats intravenously or orally with or without concomitant EGCG. Polymerase Chain Reaction and Western Blot were used to determine the effect of EGCG on drug-metabolizing enzymes (DMEs), drug transporters (DTs) and nuclear receptors (NRs). RESULTS: The Cmax and AUC of TAC were reduced, and V/F and CL/F of TAC were enhanced after co-administration of EGCG. EGCG increased the Cmax, AUC of CsA at 3 ~ 30 mg∙kg-1 dosages, while decreased those parameters at the dosage of 100 mg∙kg-1. EGCG inhibited the mRNA and protein expressions of DMEs and DTs, such as CYP3A1, A2, UGT1A1, Mdr1 and Mrp2, but upregulated the expressions of Car, Pxr and Fxr. CONCLUSIONS: These results revealed consumption of high dose EGCG may cause a significant alteration in pharmacokinetics of TAC and distribution/elimination profiles of CsA through the regulation of DMEs, DTs and NRs.

Extracts from Almond (Terminalia catappa) leaf and stem bark mitigate the activities of crucial enzymes and oxidative stress associated with hypertension in cyclosporine A-stressed rats.

We investigated the effect of extracts from the leaf (ALE) and stem bark (ABE) of Almond tree on activities of some crucial enzymes [angiotensin-1 converting enzyme (ACE), arginase, acetylcholinesterase (AChE), phosphodiesterase-5 (PDE-5), adenosine deaminase (ADA), superoxide dismutase (SOD), catalase], and thiobarbituric acid reactive species (TBARS) associated with hypertension in normal adult male Wistar albino rats and Cyclosporine A (CsA)-stressed rats. The result revealed that CsA-stressed rats treated with captopril and extracts (ALE and ABE) had lowered ACE, arginase, AChE, PDE-5, ADA activities, and TBARS level, coupled with improved SOD and catalase activities compared with untreated CsA-stressed rats, which had reversed these biochemicals compared to normal rats. This suggests that the extracts could be explored to suppress hypertension and other cardiac injury known with CsA treatment; the potentials that could be linked with the constituent polyphenols. However, further studies including blood pressure should be determined to ascertain this claim. PRACTICAL APPLICATIONS: Drug-induced cardiotoxicity, hypertension, and organ damage are among the most common side effects of pharmaceutics. Therefore, it becomes imperative to find natural, effective, and alternative therapy with little or no side effect to combat drug toxicity. The use of Almond (leaf and stem bark) in folklore for the treatment/management of hypertension and other heart-related diseases without full scientific basis is on the increase. Hence, this study provides some biochemical evidences on the effect of Almond leaf and stem back extracts on crucial enzymes and oxidative stress markers involve in the incidence of hypertension in the course of Cyclosporine A administration. The findings of this study indicated that the studied plant materials could be promoted as nutraceutical agents to neutralize drug-induced cardiac injury and hypertension.

Dextran sulfate sodium-induced colitis and ginseng intervention altered oral pharmacokinetics of cyclosporine A in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Application of cyclosporine A (CsA) as a rescue treatment in acute severe ulcerative colitis (UC) is limited by its narrow therapeutic window and great interpatient variability. As a substrate of cytochrome P450 3A enzyme (CYP3A) and P-glycoprotein (P-gp), the oral pharmacokinetics of CsA is susceptible to disease status and concomitant medications. Combined treatment with ginseng, a famous medicinal herb frequently prescribed for ameliorating abnormal immune response in many diseases including UC, showed immunologic safety in CsA-based immunosuppression. AIM OF THE STUDY: Since the therapeutic levels of CsA can be achieved within 24 h, this study first assessed the impact of acute colitis and ginseng intervention on the single oral dose pharmacokinetics of CsA and explored the underlying mechanisms in dextran sulfate sodium (DSS)-induced colitis rats and Caco-2 cells. MATERIALS AND METHODS: Rats received drinking water (normal group), 5% DSS (UC group), or 5% DSS plus daily oral ginseng extract (GS+UC group). On day 7, GS+UC group only received an oral dose of CsA (5 mg/kg), while animals of normal or UC group received an oral, intravenous (1.25 mg/kg), or intraperitoneal dose of CsA (1.25 mg/kg), respectively. Blood, liver/intestine tissues and fecal samples were collected for determining CsA and main hydroxylated metabolite HO-CsA or measuring hepatic/intestinal CYP3A activity. Caco-2 cells were incubated with gut microbial culture supernatant (CS) of different groups or ginseng (decoction or polysaccharides), and then CYP3A, P-gp and tight junction (TJ) proteins were determined. RESULTS: Oral CsA exhibited enhanced absorption, systemic exposure and tissue accumulation, and lower fecal excretion, while intravenous or intraperitoneal CsA showed lower systemic exposure and enhanced distribution, in colitis rats. Diminished intestinal and hepatic P-gp expression well explained the changes with DSS-induced colitis. Moreover, blood exposures of HO-CsA in both normal and colitis after oral dosing were significantly higher than intravenous/intraperitoneal dosing, supporting the dominant role of intestinal first-pass metabolism. Interestingly, colitis reduced CYP3A expression in intestine and liver but only potentiated intestinal CYP3A activity, causing higher oral systemic exposure of HO-CsA. Oral ginseng mitigated colitis-induced down-regulation of CYP3A and P-gp expression, facilitated HO-CsA production, biliary excretion and colonic sequestration of CsA, while not affected CsA oral systemic exposure. In Caco-2 cells, gut microbial CS from both colitis and GS+UC group diminished P-gp function, while ginseng polysaccharides directly affected ZO-1 distribution and suppressed TJ proteins expression, explaining unaltered oral CsA systemic exposure. CONCLUSIONS: DSS-induced colitis significantly altered oral CsA disposition through regulating intestinal and hepatic P-gp and CYP3A. One-week ginseng treatment enhanced colonic accumulation while not altered the systemic exposure of CsA after single oral dosing, indicating pharmacokinetic compatibility between the two medications.

Low Content of Cyclosporine A and Its Metabolites in the Colostrum of Post-Transplant Mothers.

The rate of post-transplant mothers who breastfeed while on immunosuppression is progressively increasing. Data on breastfeeding while on cyclosporine-based regimens are limited. Therefore, we assessed the amount of cyclosporine and its metabolites that might be ingested by a breastfed infant by measuring the concentration of cyclosporine and its metabolites in the colostrum of seven post-transplant mothers. The mean concentration of cyclosporine in the colostrum was 22.40 ± 9.43 mcg/L, and the estimated mean daily dose of the drug was 1049.22 ± 397.41 ng/kg/24 h. Only three metabolites (AM1, DHCsA, and THCsA) had mean colostrum amounts comparable to or higher than cyclosporine itself, with the daily doses being 468.51 ± 80.37, 2757.79 ± 1926.11, and 1044.76 ± 948.56 ng/kg/24 h, respectively. Our results indicate a low transfer of cyclosporine and its metabolites into the colostrum in the first two days postpartum and confirm the emerging change to the policy on breastfeeding among post-transplant mothers. A full assessment of the safety of immunosuppressant exposure via breastmilk will require further studies with long-term follow-ups of breastfed children.

Cyclosporine A induces testicular injury via mitochondrial apoptotic pathway by regulation of mir-34a and sirt-1 in male rats: The rescue effect of curcumin.

Testicular damage contributes to cyclosporine A (CsA) induced male infertility. However, the exact underlying molecular mediators involved in CsA-induced testis disorder remains unclear. The present study aimed to characterize the role of mir-34a/sirt-1 in CsA induced testicular injury alone or in combination with curcumin. A total of twenty-eight male Wistar rats were subdivided into four groups: control (Con), sham, cyclosporine A (CsA), cyclosporineA + curcumin (CsA + cur). The animals received cyclosporine A (30 mg/kg) and curcumin (40 mg/kg) for 28 days by oral gavage. At the end of the experiment, CsA administration significantly resulted in a decrease in testis weight and testis coefficient. The molecular analysis demonstrated that CsA exposure increased 8-OHdg and Nox4 protein contents in the testis tissue. TUNEL staining indicated that CsA caused the number of apoptotic cells to increase in the testes of male rats. In addition, exposure to CsA resulted in an increased expression of Bax, and a decreased expresion in that of Bcl-2, with a concomitant up-regulation of the Bax/Bcl-2, c-Caspase-3/p-Caspase-3 ratio and cytochrome c level. Meanwhile, exposure to CsA increased the expression of mir-34a and decreased sirt-1 protein level in the testis tissue samples compared to the control group. Taken together, our findings suggested that CsA can cause damage to testicular germ cells via oxidative stress and mitochondrial apoptotic pathway, and probably mir-34a/sirt-1 play a crucial role in this process. It also demonstrates that these negative effects of CsA can be reduced by using curcumin as an antioxidant and anti-inflammatory agent.

Costus afer leaf extract protects against testicle damage caused by cyclosporine A in adult male Wistar rats through an antioxidant mechanism.

Cyclosporine A is one of the most widely used drugs in organ transplant and oncology patients. But its use is accompanied by many toxicities. This study aimed to investigate the possible protective effect of Costus afer (C. afer) leaf extract on cyclosporine A-induced testicular toxicity. This study was carried out on 40 adult male Wistar rats were divided into four groups: control, C. afer, cyclosporine A and cyclosporine A+ C. afer groups. The investigations include genital weight, sperm count and characters, serum luteinising hormone (LH) and testosterone, testicular tissue contents of reduced glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSHPx) and lipid peroxidation (MDA). Besides, a histopathological examination of testicular tissue stained with haematoxylin and eosin (H & E) was performed. Cyclosporine A+ C. afer group showed a significant increase in the genital weight, serum testosterone, sperm count, motility and viability. Besides, the extract significantly decreased testicular content of MDA and increased SOD, CAT and GSHPx. C. afer coadministration significantly decreased serum LH and sperm abnormalities and protected against testicular histopathological alterations. The extract showed a protective effect against testicular toxicity associated with cyclosporine A and that was through an antioxidant mechanism.

Evaluation of in vitro antileishmanial efficacy of cyclosporin A and its non-immunosuppressive derivative, dihydrocyclosporin A.

BACKGROUND: New therapeutic drugs are urgently needed against visceral leishmaniasis because current drugs, such as pentavalent antimonials and miltefosine, produce severe side effects and development of resistance. Whether cyclosporine A (CsA) and its derivatives can be used as therapeutic drugs for visceral leishmaniasis has been controversial for many years. METHODS: In this study, we evaluated the efficacy of CsA and its derivative, dihydrocyclosporin A (DHCsA-d), against promastigotes and intracellular amastigotes of Leishmania donovani. Sodium stibogluconate (SSG) was used as a positive control. RESULTS: Our results showed that DHCsA-d was able to inhibit the proliferation of L. donovani promastigotes (IC50: 21.24 µM and 12.14 µM at 24 h and 48 h, respectively) and intracellular amastigotes (IC50: 5.23 µM and 4.84 µM at 24 and 48 h, respectively) in vitro, but CsA treatment increased the number of amastigotes in host cells. Both DHCsA-d and CsA caused several alterations in the morphology and ultrastructure of L. donovani, especially in the mitochondria. However, DHCsA-d showed high cytotoxicity towards cells of the mouse macrophage cell line RAW264.7, with CC50 values of 7.98 µM (24 h) and 6.65 µM (48 h). Moreover, DHCsA-d could increase IL-12, TNF-α and IFN-γ production and decrease the levels of IL-10, IL-4, NO and H2O2 in infected macrophages. On the contrary, CsA decreased IL-12, TNF-α, and IFN-γ production and increased the levels of IL-10, IL-4, NO and H2O2 in infected macrophages. The expression of L. donovani cyclophilin A (LdCyPA) in promastigotes and intracellular amastigotes and the expression of cyclophilin A (CyPA) in RAW 264.7 cells were found to be significantly downregulated in the CsA-treated group compared to those in the untreated group. However, no significant changes in LdCyPA and CyPA levels were found after DHCsA-d or SSG treatment. CONCLUSIONS: Our findings initially resolved the dispute regarding the efficacy of CsA and DHCsA-d for visceral leishmaniasis treatment. CsA showed no significant inhibitory effect on intracellular amastigotes. DHCsA-d significantly inhibited promastigotes and intracellular amastigotes, but it was highly cytotoxic. Therefore, CsA and DHCsA-d are not recommended as antileishmanial drugs.

Berberine combined with cyclosporine A alleviates acute graft-versus-host disease in murine models.

Graft-versus-host disease (GVHD) causes significant mortality after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Berberine (BBR) is primarily used to alleviate inflammation caused by autoimmune disorders. Herein the effect of BBR and cyclosporine A (CsA) on GVHD prevention in murine models is explored. Acute GVHD was induced by total body irradiation and tail vein injection with the mixture of bone marrow cells and spleen lymphocytes. Then models were treated with BBR (10 mg/kg), CsA (5 mg/kg) or the combination of BBR and CsA (10 mg/kg and 5 mg/kg) once a day for 10 days. The survival rate, weight loss and GVHD index were monitored. Then the histological changes, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, apoptosis and the levels of inflammatory cytokines, oxidative stress and nuclear factor-κB (NF-κB) signaling in liver and intestine were analyzed. Moreover, the levels of inflammatory cytokines and oxidative stress, and the count of T helper 1 (Th1) cells and Th17 cells in peripheral blood were determined. The results showed that BBR reduced GVHD-induced weight loss and GVHD index scores, attenuated liver and intestinal injury, and inhibited ALT and AST activities, inflammation, oxidative stress and NF-κB activation in liver and intestine. Additionally, BBR inhibited inflammation and reduced Th1 cell counts but had no effect on Th17 cell counts. Interestingly, the concomitant therapy of BBR and CsA was more potent than either BBR or CsA and effectively elevated the survival rate of GVHD models. This present study provides a new therapeutic strategy for alleviation of acute GVHD.

Protecting Intestinal Microenvironment Alleviates Acute Graft-Versus-Host Disease.

Acute gut graft-versus-host disease (aGVHD) is a leading threat to the survival of allogeneic hematopoietic stem cell transplantation (allo-HSCT) recipients. Abnormal gut microbiota is correlated with poor prognosis in allo-HSCT recipients. A disrupted intestinal microenvironment exacerbates dysbiosis in GVHD patients. We hypothesized that maintaining the integrity of the intestinal barrier may protect gut microbiota and attenuate aGVHD. This hypothesis was tested in a murine aGVHD model and an in vitro intestinal epithelial culture. Millipore cytokine array was utilized to determine the expression of proinflammatory cytokines in the serum. The 16S rRNA sequencing was used to determine the abundance and diversity of gut microbiota. Combining Xuebijing injection (XBJ) with a reduced dose of cyclosporine A (CsA) is superior to CsA alone in improving the survival of aGVHD mice and delayed aGVHD progression. This regimen also reduced interleukin 6 (IL-6) and IL-12 levels in the peripheral blood. 16S rRNA analysis revealed the combination treatment protected gut microbiota in aGVHD mice by reversing the dysbiosis at the phylum, genus, and species level. It inhibited enterococcal expansion, a hallmark of GVHD progression. It inhibited enterococcal expansion, a hallmark of GVHD progression. Furthermore, Escherichia coli expansion was inhibited by this regimen. Pathology analysis revealed that the combination treatment improved the integrity of the intestinal tissue of aGVHD mice. It also reduced the intestinal permeability in aGVHD mice. Besides, XBJ ameliorated doxorubicin-induced intestinal epithelial death in CCK-8 assay. Overall, combining XBJ with CsA protected the intestinal microenvironment to prevent aGVHD. Our findings suggested that protecting the intestinal microenvironment could be a novel strategy to manage aGVHD. Combining XBJ with CsA may reduce the side effects of current aGVHD prevention regimens and improve the quality of life of allo-HSCT recipients.

Clinical effect of sodium hyaluronate and cyclosporine A eye drops for combination treatment of mixed dry eye syndrome / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To analyze the efficacy of sodium hyaluronate and cyclosporine A eye drops in treating patients with mixed dry eye disease. <p>METHODS: Among patients diagnosed with dry eye who presented to our hospital between February 2018 to February 2019, 60(120 eyes)cases were divided into 30(60 eyes)in each control and treatment group by random number table method. Both groups received routine treatment with the methods of eyelid hot compress cleaning and meibomian gland massage. The treatment group received combined application of sodium hyaluronate eye drop(0.3%)and cyclosporine A eye drop(1%), while control group received carbomer eye drops used alone at four times daily. And dry eye symptom score, Schirmer I test(SⅠt), tear film break-up time(BUT)and corneal fluorescein staining(CFS)results were obtained before treatment, at the 1 and 3mo after initiation of treatment. <p>RESULTS: No statistically differences were observed in any of the indexes between the control group(dry eye symptom score: 13.52±2.15, SⅠt: 5.22±2.23, BUT: 5.02±1.58, CFS:2.82±0.81)and the treatment group(dry eye symptom score: 13.75+3.05, SⅠt: 5.54+2.89, BUT: 5.14+1.84, CFS: 2.73±0.45)before initiating treatment. One month later, the dry eye symptom score of the control group(12.22±2.64)and the treatment group(11.42±2.06)improved after treatment; the SⅠt of the control group(7.94±2.15)and the treatment group(8.63±2.78)also improved after treatment, and result of the treatment group was better than that of the control group, and the difference was statistically significant; the BUT of the control group(5.32±1.34)and the treatment group(5.46±1.45)were better after treatment, but the difference was not statistically significant. After 3mo treatment, the dry eye symptom score, SⅠt, BUT and CFS of the control group were 11.57±2.98, 8.44±2.35, 5.92±1.75, 1.92±0.44, respectively, and the dry eye symptom score, SⅠt, BUT and CFS of the control group were 9.23±2.34, 10.45±2.65, 5.92±1.75, 8.69±1.78, 1.59±0.79, respectively(<i>P</i><0.05).<p>CONCLUSIONS: Combination therapy of sodium hyaluronate eye drop and cyclosporine A eye drops treatments are effective for the treatment of mixed dry eye syndrome.

Clinical effect of sodium hyaluronate and cyclosporine A eye drops for combination treatment of mixed dry eye syndrome / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To analyze the efficacy of sodium hyaluronate and cyclosporine A eye drops in treating patients with mixed dry eye disease. <p>METHODS: Among patients diagnosed with dry eye who presented to our hospital between February 2018 to February 2019, 60(120 eyes)cases were divided into 30(60 eyes)in each control and treatment group by random number table method. Both groups received routine treatment with the methods of eyelid hot compress cleaning and meibomian gland massage. The treatment group received combined application of sodium hyaluronate eye drop(0.3%)and cyclosporine A eye drop(1%), while control group received carbomer eye drops used alone at four times daily. And dry eye symptom score, Schirmer I test(SⅠt), tear film break-up time(BUT)and corneal fluorescein staining(CFS)results were obtained before treatment, at the 1 and 3mo after initiation of treatment. <p>RESULTS: No statistically differences were observed in any of the indexes between the control group(dry eye symptom score: 13.52±2.15, SⅠt: 5.22±2.23, BUT: 5.02±1.58, CFS:2.82±0.81)and the treatment group(dry eye symptom score: 13.75+3.05, SⅠt: 5.54+2.89, BUT: 5.14+1.84, CFS: 2.73±0.45)before initiating treatment. One month later, the dry eye symptom score of the control group(12.22±2.64)and the treatment group(11.42±2.06)improved after treatment; the SⅠt of the control group(7.94±2.15)and the treatment group(8.63±2.78)also improved after treatment, and result of the treatment group was better than that of the control group, and the difference was statistically significant; the BUT of the control group(5.32±1.34)and the treatment group(5.46±1.45)were better after treatment, but the difference was not statistically significant. After 3mo treatment, the dry eye symptom score, SⅠt, BUT and CFS of the control group were 11.57±2.98, 8.44±2.35, 5.92±1.75, 1.92±0.44, respectively, and the dry eye symptom score, SⅠt, BUT and CFS of the control group were 9.23±2.34, 10.45±2.65, 5.92±1.75, 8.69±1.78, 1.59±0.79, respectively(<i>P</i><0.05).<p>CONCLUSIONS: Combination therapy of sodium hyaluronate eye drop and cyclosporine A eye drops treatments are effective for the treatment of mixed dry eye syndrome.

Cyclosporine A promotes the therapeutic effect of mesenchymal stem cells on transplantation reaction.

The successful application of mesenchymal stem cells (MSCs) remains a major challenge in stem cell therapy. Currently, several in vitro studies have indicated potentially beneficial interactions of MSCs with immunosuppressive drugs. These interactions can be even more complex in vivo, and it is in this setting that we investigate the effect of MSCs in combination with Cyclosporine A (CsA) on transplantation reaction and allogeneic cell survival. Using an in vivo mouse model, we found that CsA significantly promoted the survival of MSCs in various organs and tissues of the recipients. In addition, compared to treatment with CsA or MSCs alone, the survival of transplanted allogeneic cells was significantly improved after the combined application of MSCs with CsA. We further observed that the combinatory treatment suppressed immune response to the alloantigen challenge and modulated the immune balance by harnessing proinflammatory CD4+T-bet+ and CD4+RORγt+ cell subsets. These changes were accompanied by a significant decrease in IL-17 production along with an elevated level of IL-10. Co-cultivation of purified naive CD4+ cells with peritoneal macrophages isolated from mice treated with MSCs and CsA revealed that MSC-educated macrophages play an important role in the immunomodulatory effect observed on distinct T-cell subpopulations. Taken together, our findings suggest that CsA promotes MSC survival in vivo and that the therapeutic efficacy of the combination of MSCs with CsA is superior to each monotherapy. This combinatory treatment thus represents a promising approach to reducing immunosuppressant dosage while maintaining or even improving the outcome of therapy.