Yan Ding

CD200R1 agonist attenuates LPS-induced inflammatory response in human renal proximal tubular epithelial cells by regulating TLR4-MyD88-TAK1-mediated NF-κB and MAPK pathway

Previous studies have revealed the anti-inflammatory effect of CD200Fc, an agonist of CD200R1 in autoimmune disease. However, little is known about its anti-inflammatory effects in kidney diseases. The aim of this study is to assess the function of CD200Fc in regulating lipopolysaccharide (LPS)-induced inflammatory response in human renal proximal tubular epithelial cells (hRPTECs) and the possible mechanisms. LPS reduced the CD200R1 expression in hRPTECs, and this effect was attenuated by CD200Fc in a dose-dependent manner. In addition, CD200Fc inhibited LPS-induced expressions of TLR4 and its adapter molecule (MyD88 and phosphorylation of TAK1), and abolished its interactions with MyD88 or TAK1 in hRPTECs cells. CD200Fc also attenuated LPS-induced phosphorylation of IκB, NF-κB-P65 translocation to nucleus, and increased phosphorylation of ERK1/2, p38 and JNK in hRPTECs. Moreover, CD200Fc suppressed the LPS-induced release of pro-inflammatory mediators in hRPTECs, including IL-1β, IL-6, IL-8, MCP-1, VCAM-1, ICAM-1, TNF-α, INF-α and INF-γ. Our results suggested that CD200Fc could inhibit the TLR4-mediated inflammatory response in LPS-induced hRPTECs, thus might be beneficial for the treatment of renal disease, such as lupus nephritis.

Roles of 1,25(OH)2D3 and Vitamin D Receptor in the Pathogenesis of Rheumatoid Arthritis and Systemic Lupus Erythematosus by Regulating the Activation of CD4+ T Cells and the PKCδ/ERK Signaling Pathway

Background/Aims: The study aims to elucidate the roles of 1,25(OH)2D3 and vitamin D receptor (VDR) in the pathogenesis of rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) by regulating the activation of CD4+ T cells and the PKCδ/ERK signaling pathway. Methods: From January 2013 to December 2015, a total of 130 SLE patients, 137 RA patients and 130 healthy controls were selected in this study. Serum levels of 1,25(OH)2D3 and VDR mRNA expression were detected by ELISA and real-time fluorescence quantitative PCR (RT-qPCR). Density gradient centrifugation was performed to separate peripheral blood mononuclear cells (PBMCs). CD4+ T cells were separated using magnetic activated cell sorting (MACS). CD4+T cells in logarithmic growth phase were collected and assigned into 9 groups: the normal control group, the normal negative control (NC) group, the VDR siRNA group, the RA control group, the RA NC group, the VDR over-expressed RA group, the SLE control group, the SLE NC group, and the VDR over-expressed SLE group. The mRNA and protein expressions of VDR, PKCδ, ERK1/2, CD11a, CD70 and CD40L were detected by RT-qPCR and Western blotting. Bisulfite genomic sequencing was conducted to monitor the methylation status of CD11a, CD70 and CD40L. Results: Compared with healthy controls, serum 1,25(OH)2D3 level and VDR mRNA expression in peripheral blood were decreased in SLE patients and RA patients. With the increase of concentrations of 1,25(OH)2D3 treatment, the VDR mRNA expression and DNA methylation levels of CD11a, CD70 and CD40L were declined, while the expressions of PKCδ, ERK1/2, CD11a, CD70 and CD40L were elevated in SLE, RA and normal CD4+T cells. Compared with the SLE contro, RA control, SLE NC and RA NC groups, the expressions of PKCδ, ERK1/2, CD11a, CD70 and CD40L decreased but DNA methylation levels of CD11a, CD70 and CD40L increased in the VDR over-expressed SLE group and VDR over-expressed RA group. However, compared with the normal control and normal NC groups, the expressions of PKCδ, ERK1/2, CD11a, CD70 and CD40L increased, but DNA methylation levels of CD11a, CD70 and CD40L decreased in the VDR siRNA group. Compared with the normal control group, the expressions of PKCδ, ERK1/2, CD11a, CD70 and CD40L increased, but DNA methylation levels of CD11a, CD70 and CD40L decreased in the SLE control and RA control groups. Conclusion: Our study provide evidence that 1,25(OH)2D3 and VDR could inhibit the activation of CD4+ T cells and suppress the immune response of SLE and RA through inhibiting PKCδ/ERK pathway and promoting DNA methylation of CD11a, CD70 and CD40L.

CSTMP Exerts Anti-Inflammatory Effects on LPS-Induced Human Renal Proximal Tubular Epithelial Cells by Inhibiting TLR4-Mediated NF-κB Pathways.

Abstract(E)-2-(2-chlorostyryl)-3,5,6-trimethylpyrazine (CSTMP), a novel stilbene derivative, have been shown to have cytoprotective effects against H2O2-induced oxidative stress in human endothelial cells. However, little is known about its anti-inflammatory effects in lupus nephritis (LN). In the present study, we investigated the anti-inflammatory effects of CSTMP on lipopolysaccharide (LPS)-induced human renal proximal tubular epithelial cells (hRPTECs) and elucidated its molecular mechanisms. CSTMP significantly attenuated the cytotoxicity and suppressed the release of proinflammatory mediators, including iNOS, COX-2, TNF-α, IL-6, IL-8, CCL-2, ICAM-1, IL-1β, and MCP-1 in LPS-induced hRPTECs. In addition, CSTMP decreased the expression of TLR4 and its adapter molecules (MyD88, phosphorylation of TAK1, TRAF6, and IRAK1) and abolished its interactions with these adapter molecules in LPS-induced hRPTECs, resulting in an inhibition of the TLR4/MyD88/TAK1/ TRAF6/IRAK1 complex. Moreover, CSTMP also attenuated phosphorylation of IκB and IKK-α/β, and P50-NF-κB and P65-NF-κB translocation to nucleus in LPS-induced hRPTECs. These findings provided new insights to understand the mode of action of CSTMP in treatment of inflammatory diseases, such as LN.

Effects of 1,25(OH)2D3 and vitamin D receptor on peripheral CD4+/CD8+ double‐positive T lymphocytes in a mouse model of systemic lupus erythematosus

This study aims to explore effects of 1,25(OH)2D3 and vitamin D receptor (VDR) on peripheral CD4+/CD8+ double‐positive (DP) T lymphocytes in systemic lupus erythematosus (SLE). MRL‐LPr/LPr mice with SLE (n = 20) and normal MRL mice (n = 20) were assigned into the control group (normal mice, without feeding with 1,25(OH)2D3), the 1,25(OH)2D3 group (SLE mice, feeding with 1,25(OH)2D3), the VDR‐knock‐in + 1,25(OH)2D3 group (SLE mice, VDR‐knock‐in, feeding with 1,25(OH)2D3) and the VDR‐knockout group (normal mice, VDR‐knockout, without feeding with 1,25(OH)2D3) (n = 10 per group). Levels of T lymphocytes were measured by flow cytometry. The mRNA and proteins expressions of inflammatory factors were measured by qRT‐PCR and ELISA. Extracellular signal‐regulated kinase‐1/2 (ERK1/2) expression was measured by Western blotting. Compared with normal mice, SLE mice showed reduced levels of CD4+, CD4+/CD8+ ratio, and DP lymphocytes. The levels of SLE‐related indicators all increased significantly, followed with severe skin ulcers and urinary system infection. With the increase in time, skin ulcers and urinary system infection were significantly improved, levels of CD4+, CD4+/CD8+ ratio, and DP lymphocytes increased, and levels of SLE‐related indicators all decreased in the 1,25(OH)2D3 group. There were no significant changes in bioindicators in the control and the VDR‐knock‐in + 1,25(OH)2D3 groups. The symptoms of SLE gradually occurred in the VDR‐knockout group. This study demonstrates that VDR and 1,25(OH)2D3 could elevate CD4+/CD8+ DP T lymphocytes and reduce expressions of inflammatory factors, thus inhibiting the development and progression of SLE.

The expression of EBV-encoded LMP1 in young patients with lupus nephritis

Hair care products are expected to wet well human hair, even when the hair is hydrophobic. Thus, wetting properties of human hair are very important, as they influence consumer satisfaction with the products. Wettability of a hair tress is an important characteristic. The wetting behavior of polymer solutions on hair is less studied than surfactant solutions. The wetting of hair tresses by aqueous solutions of commercially available polymers AculynTM 22 (A22) and AculynTM 33 (A33) has been investigated. Both experimental studies and numerical simulations of behavior of polymer solutions and foams on tresses of human hair has been investigated including drainage of foams produced from solutions of those polymers and interaction of foams with hair stresses are presented. Both A22 and A33 solutions demonstrate well pronounced shear thinning behavior. Initial contact angle of the A22 and A33 solutions on undamaged hair tresses is about 100 o. The A22 droplets remained on the hair tress after spreading for at least half an hour. However, a fast penetration of the A33 droplet inside the hair tresses was observed when advancing contact angle in the course of spreading reaches a critical value of about (60 o). Pure solutions of A22 and A33 have higher initial contact angle and longer penetration time on hair tresses compared with the solutions containing i-propanol or sodium dodecyl sulphate. The results demonstrate that wetting kinetics of the polymer solution on hair tresses drastically different depending on the formulation and can vary from a rapid imbibition to a spreading only.One of the major disease manifestations of systemic lupus erythematosus (SLE) is lupus nephritis (LN), and the underlying mechanisms are not yet understood. Epstein-Barr virus (EBV) reactivation was associated with the induction of SLE, with EBV-encoded latent membrane protein1 (LMP1) plays a vital role in this process. Although it was reported that LN was associated with LMP1, most of these results are from patients with ages differed greatly (range, 10-56 years). Given the increased prevalence of EBV infection in young patients, we focused on the association of LN and LMP1 expression in the renal tissues of young patients (range, 6-16 years) in this study. We found that the positive rate of LMP1 in the renal tissues was significantly higher in patients with LN compared with control (P<0.001), which is consistent with the previous reports. The positive rates of LMP1 were similar between the patients of initial onset and relapse, and there was no detectable difference between the patients with and without concurrent infection (P>0.05). However, we reported for the first time about the positive correlation of LMP1 with classification of LN. The proportion of young patients positive for anti‑Sm antibody was significantly higher in the LMP1 positive group compared with the LMP1 negative control (P>0.05). These results indicate that EBV infection in the renal of young patients may lead to the increased severity of LN, and the expression of anti-Sm is likely contributed to this process.Human hair is a nanocomposite biological fiber. Maintaining the health, feel, shine, color, softness, and overall aesthetics of the hair is highly desired. Hair care products such as shampoos and conditioners, along with damaging processes such as chemical dyeing and permanent wave treatments, affect the maintenance and grooming process and are important to study because they alter many hair properties. Nanoscale characterization of the cellular structure, mechanical properties, and morphological, frictional, and adhesive properties (tribological properties) of hair are essential to evaluate and develop better cosmetic products, and to advance the understanding of biological and cosmetic science. The tensile response of hair is of considerable interest. Another property of interest is the surface charge of hair, which has a significant effect on manageability, feel, and appearance. For this reason, controlling charge buildup to improve these factors is an important issue in the commercial hair care industry. The atomic/friction force microscope (AFM/FFM) and nanoindenter have recently become important tools for studying the micro/nanoscale properties of human hair. In this talk, we present a comprehensive review of the cellular structural, nanomechanical, and nanotribological properties of various hair and skin as a function of ethnicity, damage, conditioning treatment, and various environments1-2. Various cellular structures of human hair and fine sublamellar structures of the cuticle are identified and studied. Nanomechanical properties such as hardness, elastic modulus, creep and scratch resistance are discussed. Nanotribological properties such as roughness, friction, and adhesion are presented, as well as investigations of conditioner distribution, thickness, and binding interactions.

Curcumin inhibited growth of human melanoma A375 cells via inciting oxidative stress

Curcumin, a polyphenol compound, possesses potent pharmacological properties in preventing cancers, which make it as a potential anti-cancer mediator. However, it is still unknown that whether Curcumin induced melanoma A375 cell was associated with oxidative stress. Here, we firstly found a fascinating result that Curcumin could reduce the proliferation and induced apoptosis of human melanoma A375 cells. Meanwhile, IC50 of Curcumin on A375 cells is 80μM at 48h. In addition, Curcumin caused oxidative stress through inducing further ROS burst, decreasing GSH, and wrecking mitochondria membrane potential (MMP), which were reversed by ROS inhibitor N-acetylcysteine (NAC). Moreover, MMP disruption led to the release of Cytochrome c from mitochondria and subsequently led to intracellular apoptosis. Furthermore, we found that ROS-dependent HIF-1α and its downstream proteins also play an important role on Curcumin induced apoptosis. In conclusion, our results shed new lights on the therapy of melanoma that Curcumin may be a promising candidate.

25-Hydroxyvitamin D-1-α-hydroxylase in apoliporotein E knockout mice: The role of protecting vascular smooth muscle cell from calcification

Previous publications widely reported that 25-hydroxyvitamin D-1-α-hydroxylase (CYP27B1) regulated the metabolism of 25-hydroxyvitamin D3, which has a close association between altered activity of vitamin D and vascular calcification has been reported in various human diseases, including chronic kidney disease, osteoporosis and atherosclerosis. Vascular calcification is a clinically significant component of atherosclerosis and may be promoted by ROS associated inflammatory. In this study, we evaluated the effect of 25-hydroxyvitamin D-1-α-hydroxylase on the atherosclerosis disease both in apolipoprotein (apo) E-/- mice and wild-type mice. We also isolated endothelial cell (ECs) and vascular smooth muscle cells (VSMCs) in aortic from the wild type mice and apoE-/- mice respectively, then investigated that after parathyroid hormone (PTH) both of the CYP27B1 and vitamin D receptor (VDR) expressions in apoE-/-EC and apoE-/-VSMC were higher than the wide-type EC and VSMCs. However, the increased proliferation and decreased apoptosis have showed in EC and VSMC compared with the cells from apo E-/- mice. Moreover, the index associated with vascular calcification such as intracellular Ca2+ concentration and alkaline phosphatase (ALP) activity have been tested and the result suggested that the levels of the former index have improved in the apoE-/-EC and apoE-/-VSMC. We got similar conclusions under the pre-treatment with 1, 25(OH) 2D3.

Identification of a gene-expression predictor for diagnosis and personalized stratification of lupus patients

Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by a wide spectrum of clinical manifestations and degrees of severity. Few genomic biomarkers for SLE have been validated and employed to inform clinical classifications and decisions. To discover and assess the gene-expression based SLE predictors in published studies, we performed a meta-analysis using our established signature database and a data similarity-driven strategy. From 13 training data sets on SLE gene-expression studies, we identified a SLE meta-signature (SLEmetaSig100) containing 100 concordant genes that are involved in DNA sensors and the IFN signaling pathway. We rigorously examined SLEmetaSig100 with both retrospective and prospective validation in two independent data sets. Using unsupervised clustering, we retrospectively elucidated that SLEmetaSig100 could classify clinical samples into two groups that correlated with SLE disease status and disease activities. More importantly, SLEmetaSig100 enabled personalized stratification demonstrating its ability to prospectively predict SLE disease at the individual patient level. To evaluate the performance of SLEmetaSig100 in predicting SLE, we predicted 1,171 testing samples to be either non-SLE or SLE with positive predictive value (97–99%), specificity (85%-84%), and sensitivity (60–84%). Our study suggests that SLEmetaSig100 has enhanced predictive value to facilitate current SLE clinical classification and provides personalized disease activity monitoring.

Chelidonine suppresses LPS-Induced production of inflammatory mediators through the inhibitory of the TLR4/NF-κB signaling pathway in RAW264.7 macrophages

Chelidonine is one of the alkaloids of Chelidonium majus, which has broad pharmacological activities, including anti-inflammatory. Despite chelidonine has been shown to exhibit anti-inflammatory activity, the molecular mechanisms are not yet fully elucidated. In this paper, we used RAW264.7 macrophages and mice to investigate the anti-inflammatory effects of chelidonine. Firstly, we found that chelidonine significantly suppressed LPS-induced the production of NO and PGE2, as well as iNOS and COX-2 mRNA and protein expression. In addition, pro-inflammatory cytokines induced by LPS, such as TNFα and IL-6 were also attenuated by chelidonine. Whats more, LPS-induced activation and degradation of IκBα followed by translocation of the p65 from the cytoplasm to the nucleus were attenuated by chelidonine. Furthermore, chelidonine even significantly inhibited TLR4 expression induced by LPS. Finally, we verified that chelidonine striking ly decreased serum TNFα, IL-6 and PGE2 levels in LPS stimulated mice. Taken together, this study demonstrated that chelidonine may suppressed the LPS-induced inflammatory response both in vitro and in vivo, which was relating to TLR4/NF-κB signaling pathway disturbed by chelidonine.