Jiangsu Key Laboratory of Molecular Medicine

Nanjing, China

Jiangsu Key Laboratory of Molecular Medicine

Nanjing, China
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Chen Y.,Nanjing University | Chen Y.,Jiangsu Key Laboratory of Molecular Medicine | Xu J.,Institute of Toxicology and Functional Assessment | Li Y.,Nanjing University | And 4 more authors.
Reproductive Toxicology | Year: 2011

The effects of chronic low-dose exposure to microcystins were preliminarily studied on sperm quality and testicular function in male mice. Microcystin-LR (MC-LR) was orally administered to male mice at 0, 1, 3.2, and 10. μg/L for 3 and 6 months. Our preliminary study found in three-month group, sperm quality declined at 3.2 and 10. μg/L doses, testosterone dropped at 10. μg/L, levels of LH and FSH increased, and Leydig cells exhibited apoptosis. Similar, but more pronounced, effects were observed in groups treated with MC-LR for 6 months. Compared to control (0. μg/L), the rate of sperm abnormality was higher and testosterone levels were lower following administration of 3.2 and 10. μg/L MC-LR and structural damage to the testis was observed with 10. μg/L dose. Thus, chronic low-dose treatment with MC-LR results in substantial toxicity to male reproduction, causing declines in sperm quality, decreased levels of serum testosterone, and injury to the testis. © 2011 Elsevier Inc.


Chen D.,Nanjing University | Su A.,Nanjing University | Fu Y.,Nanjing University | Wang X.,Nanjing University | And 6 more authors.
Antiviral Research | Year: 2015

Herpes simplex virus types 1 and 2 (HSV-1 and -2) are highly prevalent in many populations and therapeutic options are limited. Both viruses can establish latency by maintaining viral genomes in neurons of sensory ganglia. Primary or recurrent HSV infections may lead to deleterious outcomes: HSV-1 infection may result in corneal blindness and encephalitis and HSV-2 infection leads to herpes genitalis. While no effective vaccine is available, acyclovir is widely used for therapy, which targets and inhibits viral DNA polymerase. Although acyclovir is of low toxicity, resistant strains arise due to persistent use, mainly in immune compromised patients. In our effort to identify new HSV inhibitory molecules, harmine was found to potently inhibit HSV infection. Harmine, a beta-carbon alkaloid with an indole core structure and a pyridine ring, is widely distributed in plants. Earlier studies showed that harmine exhibited pharmacological activities such as antifungal, antimicrobial, antitumor, antiplasmodial and antioxidants. In the current study, we showed that harmine was a potent inhibitor of HSV-2 infection in vitro assays with EC50 value at around 1.47 μM and CC50 value at around 337.10 μM. The HSV RNA transcription, protein synthesis, and virus titers were reduced by the presence of harmine in a dose dependent manner. Further study on the mechanism of the anti-HSV activity showed that harmine blocked HSV-induced ROS production and the upregulated cytokine/chemokine expression, but our evidence showed that the inhibition of viral replication was unlikely mediated by the blocking of ROS production. We demonstrated that harmine significantly reduced HSV-2-induced NF-κB activation, as well as IκB-α degradation and p65 nuclear translocation. We found that harmine also inhibited HSV-2-mediated p38 kinase and c-Jun N-terminal kinases (JNK) phosphorylation. © 2015, Elsevier B.V. All rights reserved.


Zhou Y.,Nanjing University | Zhou Y.,Jiangsu Key Laboratory of Molecular Medicine | Yuan J.,Nanjing University | Yuan J.,Jiangsu Key Laboratory of Molecular Medicine | And 4 more authors.
Toxicology Letters | Year: 2012

Microcystin-leucine arginine (MC-LR), a cyclic heptapeptide produced by several bloom-forming cyanobacteria, has strong reproductive toxicity. We examined whether MC-LR could enter spermatogonia and investigated the toxic effects of MC-LR on spermatogonia in vitro. Multispecific organic anion-transporting polypeptides (Oatps), which transported MCs, were screened as well. Spermatogonia were exposed to 0, 0.5, 5, 50, and 500nmol/L (nM) MC-LR for 6h. Cell viability and total antioxidant capacity significantly decreased, meanwhile, the ratio of apoptotic cells, reactive oxidative species (ROS) production, mitochondrial membrane potential (MMP), and intracellular free Ca 2+ increased after exposure to 5nM and higher concentrations of MC-LR. MC-LR can immigrate into spermatogonia. At least 5 Oatps (Oatp1a5, -3a1, -6b1, -6c1, and -6d1) were detected at the mRNA level in spermatogonia, and the expression of these Oatps was affected by MC-LR, especially Oatp3a1. This study demonstrated that MC-LR can be transported into spermatogonia and leads to cytotoxicity. © 2012 Elsevier Ireland Ltd.


Fan H.,Nanjing University | Zhao G.,Nanjing University | Liu L.,Nanjing University | Liu F.,Nanjing University | And 6 more authors.
Cellular and Molecular Immunology | Year: 2012

Mesenchymal stem cells (MSCs) have been used experimentally for treating inflammatory disorders, partly due to their immunosuppressive properties. Although interleukin-1β (IL-1β) is one of the most important inflammatory mediators, growing evidence indicates that IL-1β signaling elicits the immunosuppressive properties of MSCs. However, it remains unclear how IL-1β signaling accomplishes this activity. Here, we focus on the therapeutic efficacy of IL-1β-primed MSCs in the dextran sulfate sodium (DSS)-induced colitis model, in addition to the underlining mechanisms. We first found that IL-1β-primed MSCs, without any observable phenotype change in vitro, significantly attenuated the development of DSS-induced murine colitis. Moreover, IL-1β-primed MSCs modulated the balance of immune cells in the spleen and the mesenteric lymph nodes (MLNs) through elevating cyclooxygenase-2 (COX-2), IL-6 and IL-8 expression and influencing the polarization of peritoneal macrophages. Importantly, IL-1β-primed MSCs possessed an enhanced ability to migrate to the inflammatory site of the gut via upregulation of chemokine receptor type 4 (CXCR4) expression. In summary, IL-1β-primed MSCs have improved efficacy in treating DSS-induced colitis, which at least partly depends on their increased immunosuppressive capacities and enhanced migration ability. © 2012 CSI and USTC. All rights reserved.


Zhao X.,Nanjing University | Liu L.,Nanjing University | Liu D.,Nanjing University | Fan H.,Nanjing University | And 4 more authors.
American Journal of Reproductive Immunology | Year: 2012

Problem: Progesterone (P4) plays a central role in the establishment and maintenance of pregnancy. It also has profound effects on the regulation of immune responses. Mesenchymal stem cells (MSCs), which are thought to have the ability to modulate immunocyte activation, are present in human endometrium and deciduas and highly express progesterone receptor (PR). Especially, during pregnancy, both P4 and MSCs are present and regulatively changed at the fetal-maternal interface, but the effect of P4 on the MSCs remains unknown. Therefore, in this study, we investigated the effects of P4 on the immunomodulatory ability of MSCs and the underlying mechanisms. Method of study: Mesenchymal stem cells were obtained from human umbilical cords. The effects of P4 on the cell morphology, phenotype, proliferation, apoptosis, and expression levels of cytokine and protein were examined. Results: Progesterone did not affect the phenotype, morphology, proliferation, and apoptosis of MSCs, but promoted the production of PGE2 and IL-6 in MSCs. Furthermore, the up-regulation of PGE2 and IL-6 was related to the activation of p38 and NF-κB. Conclusions: Progesterone enhances immunomodulatory function of MSCs through up-regulation of PGE2 and IL-6. The data suggest that P4-regulated MSCs may play a crucial role in modulating the local immune balance of fetal-maternal interface. © 2012 John Wiley & Sons A/S.


Chen S.,Nanjing Medical University | Zhao G.,Nanjing Medical University | Miao H.,Nanjing University | Tang R.,Nanjing University | And 6 more authors.
FEBS Letters | Year: 2015

Mesenchymal stem cells (MSCs) play an important role in the pathology of preeclampsia (PE). Our previous microarray analysis found that microRNA-494 (miR-494) is highly expressed in decidua-derived MSCs (dMSCs) from PE. We hypothesized that aberrant expression of miR-494 in dMSCs is involved in PE development. In the present study, we found that miR-494 arrests G1/S transition in dMSCs by targeting CDK6 and CCND1. We also found that supernatant from miR-494-overexpressing dMSCs reduces HTR-8/SVneo migration and impairs HUVEC capillary formation by suppressing VEGF. Taken together, we report an unrecognized mechanism of miR-494 affecting dMSC proliferation and function in the pathology of PE. © 2015 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.


Fan H.,Nanjing University | Liu F.,Nanjing University | Dong G.,Nanjing University | Ren D.,Nanjing University | And 6 more authors.
Cell Death and Disease | Year: 2014

B-cell abnormality including excessive activation and lymphopenia is a central feature of systemic lupus erythematosus (SLE). Although activation threshold, auto-reaction and death of B cells can be affected by intrinsical and/or external signaling, the underlying mechanisms are unclear. Herein, we demonstrate that co-activation of Toll-like receptor 7 (TLR7) and B-cell receptor (BCR) pathways is a core event for the survival/dead states of B cells in SLE. We found that the mortalities of CD19+CD27- and CD19+IgM+ B-cell subsets were increased in the peripheral blood mononuclear cells (PBMCs) of SLE patients. The gene microarray analysis of CD19+ B cells from active SLE patients showed that the differentially expressed genes were closely correlated to TLR7, BCR, apoptosis, necroptosis and immune pathways. We also found that co-activation of TLR7 and BCR could trigger normal B cells to take on SLE-like B-cell characters including the elevated viability, activation and proliferation in the first 3 days and necroptosis in the later days. Moreover, the necroptotic B cells exhibited mitochondrial dysfunction and hypoxia, along with the elevated expression of necroptosis-related genes, consistent with that in both SLE B-cell microarray and real-time PCR verification. Expectedly, pretreatment with the receptor-interacting protein kinase 1 (RIPK1) inhibitor Necrostatin-1, and not the apoptosis inhibitor zVAD, suppressed B-cell death. Importantly, B cells from additional SLE patients also significantly displayed high expression levels of necroptosis-related genes compared with those from healthy donors. These data indicate that co-activation of TLR7 and BCR pathways can promote B cells to hyperactivation and ultimately necroptosis. Our finding provides a new explanation on B-cell lymphopenia in active SLE patients. These data suggest that extrinsic factors may increase the intrinsical abnormality of B cells in SLE patients. © 2014 Macmillan Publishers Limited. All rights reserved.


Liu L.,Nanjing University | Zhao G.,Nanjing University | Fan H.,Nanjing University | Zhao X.,Nanjing University | And 5 more authors.
PLoS ONE | Year: 2014

Pre-eclampsia (PE) is thought to be a pregnancy-induced autoimmune disease. Despite several strategies carried out for targeting specific factors relevant to its pathogenesis, PE remains potentially fatal to some patients. Here, we reported a way to isolate mesenchymal stem cells (MSCs) from decidua. The MSCs not only exhibited differentiation and self-renewal capacities, they also possessed immunomodulatory functions and secreted some soluble mediators including IL-6, TGF-β, IDO, VEGF and COX-2. Most importantly, the MSCs were specifically provided with the ability to suppress T cells proliferation by IDO in response to inflammatory cytokine IFN-γ. Moreover, we developed a Th1 cell-induced PE mouse model which displayed a high level of pathogenesis factor TNF-α. Strikingly, MSCs-based therapy significantly ameliorated both clinical and histopathological severity of PE symptoms including decreasing the blood pressure and proteinuria, suppressing glomerulonephritis, protecting the feto-placental development. The therapy also reversed abnormal TNF-α expression in uterine and splenic lymphocytes. These data suggest that MSCs may ameliorate Th1-induced PE-like symptoms in mice via the suppression of TNF-α and MSCs-based therapy may provide a potential novel method for PE. © 2014 Liu et al.


Wang Y.,Nanjing University | Fan H.,Nanjing University | Zhao G.,Nanjing Medical University | Liu D.,Nanjing University | And 5 more authors.
FEBS Journal | Year: 2012

Pre-eclampsia is thought to be a systemic disease of maternal endothelial cell dysfunctions. miRNAs regulate various basic biological functions in cells, including stem cells. Mesenchymal stem cells exist in almost all tissues and are the key cellular source for tissue repair and regeneration. Our aims are to investigate whether miRNAs regulate MSCs in fetal-maternal interfaces to influence the pathogenesis of pre-eclampsia. The differential expression of miRNAs in decidua-derived mesenchymal stem cells of all patients with severe pre-eclampsia (n = 20) and normal groups (n = 20) was first screened by microarray analysis and validated by quantitative real-time PCR analysis. The integrated bioinformatics analysis showed that miR-16 showed the highest number of connections in the miRNA GO network and the miRNA gene network. Moreover, over-expressed miR-16 inhibited the proliferation and migration of decidua-derived mesenchymal stem cells and induced cell-cycle arrest by targeting cyclin E1. Interestingly, over-expression of miR-16 by decidua-derived mesenchymal stem cells reduced the ability of human umbilical vein endothelial cells to form blood vessels and reduced the migration of trophoblast cells. Furthermore, decidua-derived mesenchymal stem cell-expressed endothelial growth factor VEGF-A was involved in migration of trophoblast cells and human umbilical vein endothelial cells as well as tube and network formation. Importantly, the levels of cyclin E1 and VEGF-A were negatively correlated with the level of miR-16 expression in decidua-derived mesenchymal stem cells from the patients with severe pre-eclampsia. Together, these data suggest that the alteration of miR-16 expression in decidua-derived mesenchymal stem cells may be involved in the development of pre-eclampsia. © 2012 FEBS.


Zhao X.,Nanjing University | Liu D.,Jiangsu Key Laboratory of Molecular Medicine | Gong W.,Nanjing University | Zhao G.,Nanjing University | And 3 more authors.
Stem Cells | Year: 2014

Mesenchymal stem cells (MSCs) are attractive candidates for clinical therapeutic applications. Recent studies indicate MSCs express active Toll-like receptors (TLRs), but their effect on MSCs and the underlying mechanisms remain unclear. In this study, we found that, after treating human umbilical cord MSCs with various TLR ligands, only TLR3 ligand, poly(I:C), could significantly increase the expression of cyclooxygenase-2 (COX-2). Furthermore, poly(I:C) could enhance MSCs' anti-inflammatory effect on macrophages. Next, we focused on the regulatory roles of microRNAs (miRNAs) in the process of poly(I:C) activating MSCs. Our experiments indicated that miR-143 expression was significantly decreased in MSCs with poly(I:C) treatment, and the expression level of miR-143 could regulate the effect of poly(I:C) on MSCs' immunosuppressive function. Subsequent results showed that the reporter genes with putative miR-143 binding sites from the transforming growth factor-b-activated kinase-1 (TAK1) and COX-2 30 untranslated regions were downregulated in the presence of miR-143. In addition, mRNA and protein expression of TAK1 and COX-2 in MSCs was also downregulated with miR-143 overexpression, suggesting that TAK1 and COX-2 are target genes of miR-143 in MSCs. Consistent with miR-143 overexpression, TAK1 interference also attenuated MSCs' immunosuppressive function enhanced by poly(I:C). Additionally, it was shown that TLR3-activated MSCs could improve survival in cecal ligation and puncture (CLP)-induced sepsis, while miR-143 overexpression reduced the effectiveness of this therapy. These results proved that poly(I:C) improved the immunosuppressive abilities of MSCs, revealed the regulatory role of miRNAs in the process, and may provide an opportunity for potential novel therapies for sepsis. © AlphaMed Press 2013.

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