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A previous study demonstrated the renal-protective effect of thalidomide (Thd) in diabetic nephropathy rats through the activation of the adenosine monophosphate-activated protein kinase (AMPK) and inhibition of the nuclear factor κB (NF-κB)/monocyte chemoattractant protein-1 (MCP-1) and transforming growth factor (TGF)-β1/mothers against decapentaplegic homolog signaling pathways. The association between AMPK inactivation and high glucose (HG)-induced meningeal cell (MC) proliferation and extracellular matrix (ECM) accumulation via NF-κB and TGF-β1 signaling remains unknown. The aim of the current study was to demonstrate the effects of Thd on cell proliferation and ECM expression in HG-cultured MCs and the underlying mechanisms. HG-cultured human MCs were treated with Thd. Cell proliferation was measured by MTT assay and quantification of cell proliferation was based on the measurement of bromodeoxyuridine incorporation. The differences in TGF-β1, fibronectin and MCP-1 protein expression levels were detected via ELISA and western blot analysis. The AMPK signaling pathway was also examined by western blot analysis in MCs. Compound C, an AMPK inhibitor and AICAR (5-aminoimidazole-4-carboxamide 1β-D-ribofuranoside), an AMPK agonist, were used to analyze the functional role of AMPK in MCs. Cell proliferation was significantly decreased in HG-cultured MCs following treatment with high concentrations of Thd (100 and 200 µg/ml) for 24 h compared with the HG-cultured MC group. Thd suppressed the inflammatory processes in HG-induced MCs. These effects were partially mediated through the activation of AMPK and inhibition of the NF-κB/MCP-1 signaling pathways. Taken together, these results suggest that Thd may have therapeutic potential in diabetic renal injury via the AMPK signaling pathway. [ABSTRACT FROM AUTHOR]

Lupus nephritis (LN) is a severe consequence of systemic lupus erythematosus (SLE) and is an important driver of morbidity and mortality in SLE. Treg cells and TIM-3 play an important role in the pathogenesis of LN. The beneficial effect of rapamycin on LN has been confirmed in both mouse models and patients, but the effect of rapamycin on Treg cells and TIM-3 is not yet completely understood. In this study, rapamycin treatment attenuated proteinuria, histological damage, and renal deposition of C3, and improved renal function. Spleen and renal draining lymph node weight and serum levels of anti-dsDNA antibodies were also improved by rapamycin. Furthermore, the frequency of Treg cells and Treg functional molecules, such as cytotoxic T cell antigen 4 (CTLA-4), interleukin 10 (IL-10), and transforming growth factor β1 (TGF-β1), increased significantly after treatment with rapamycin in MRL/lpr mice. We also found that expression of TIM-3 was significantly decreased in CD4+ T cells and Treg cells in mice treated with rapamycin. In summary, the study demonstrated that rapamycin treatment induced preferential expansion of CD4+CD25+Foxp3+ Tregs with increased expression of CTLA-4, IL-10, and TGF-β1, and decreased TIM-3 expression, thereby ameliorating lupus nephritis in the MRL/lpr mouse model., Competing Interests: The authors declare no conflict of interest., (Copyright © 2022 Termedia.)

Objective: To explore the protective effects of leflunomide (A771726) on the expression of podocalyxin, NF-κB and matrix metalloproteinase-9 (MMP-9) in podocytes exposed to high glucose environment and elucidate its mechanism., Methods: Podocytes were cultured in high glucose. And the altered expressions of podocyte protein podocalyxin were detected by Western blotting at different timepoints. Then podocytes were divided into 4 groups of normal glucose control, leflunomide, high glucose and hypertonic control. The expression level of podocalyxin protein in each group was detected by Western blotting. And NF-κB p65 and phosphorylation of NF-κB p65 (P-NF-κBp65) in podocytes cultured in high glucose were detected at different timepoints. And then the podocytes were divided into 5 groups of normal glucose, mannitol, hypertonic control, high glucose, leflunomide and PDTC (NF-κB blocker). And the expressions of MMP-9 protein in these groups were also detected by Western blotting., Results: In the high glucose environment, the expression of podocalyxin declined instantly. Compared with the high-glucose group, the podocalyxin expression of the leflunomide group was significantly higher than the high glucose group (0.46 ± 0.04 vs 0.13 ± 0.03, P < 0.05). After 30-minute stimulation by high glucose, the activation of NF-κB started and the expression of P-NF-κBp65 protein increased. Such activities peaked at 60 minutes and reverted to a basic level after 6 hours. Compared with the high glucose group, the expressions of MMP-9 in PDTC and leflunomide groups were significantly lower than the high glucose group. And the differences were statistically significant (0.71 ± 0.01, 0.64 ± 0.03 vs 1.64 ± 0.03, both P < 0.05)., Conclusions: Leflunomide has protective effects on podocytes in high glucose. And its mechanism is possibly due to a lowered expression of MMP-9 through an inhibition of NF-κB activation.

Objective: To explore the effects of leflunomide active metabolite A771726 on high glucose-induced podocyte cytoskeleton and its possible signaling pathway., Methods: The conditionally immortal human glomerular podocytes were divided into normal glucose (NG), mannitol (MA), high glucose (HG), high glucose with PDTC (pyrrolidine dithiocarbamate, a NF-κBp65 inhibitor) and high glucose with active leflunomide metabolite A771726 groups. Western blot was used to measure the ratio of p-NF-κBp65 to total NF-κBp65. And the protein and mRNA expressions of NF-κBp65, TRPC6 and nephrin were detected by Western blot and reverse transcription polymerase chain reaction (PCR). Immunofluorescence staining was used to detect the changes in the skeleton of podocyte., Results: (1) Podocytes with high glucose could activate the NF-κBp65 signaling pathway. There was a significant increase of p-NF-κBp65 protein at 60 min versus 0 min (1.20 ± 0.04 vs 0.79 ± 0.02, P < 0.01). Little activation of the pathways was observed in groups NG and MA. The up-regulated protein expression of p-NF-κBp65 induced with high glucose was significantly inhibited by PDTC and A771726 (both P < 0.05). The difference of NF-κBp65 mRNA expression was not statistically significant between the groups (all P > 0.05). (2) High glucose-induced podocyte activated the NF-κBp65 signaling path. Its downstream TRPC6 mRNA and protein expression significantly increased than NG while nephrin became down-regulated more than NG. PDTC and A771726 inhibited the high expression of TRPC6 while the expression of nephrin was elevated (all P < 0.05). (3) Immunofluorescent assay of high glucose-induced podocyte cytoskeleton showed disorderly F-actin and a disappearance of tensile fiber after 72 h., Conclusion: Active leflunomide metabolite A771726 may protect podocytes through blocking the high glucose-induced signaling pathway of NF-κBp65.

Background: It is necessary to develop some innovative methods to reveal and discover the novel (SLE)-related protein molecules. In the present study, matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) was employed to detect the differential expression of serum polypeptides in the patients with systemic lupus erythematosus (SLE) presenting with a family history or complicating with kidney injury so as to identify the proteins associated with the genetic factors and kidney injury in SLE., Methods: The subjects recruited were divided into four groups, that is, a group of SLE patients with both family history and kidney injury, a group of SLE patients with only kidney injury but no family history, a group of SLE patients with neither family history nor kidney injury, and a control group consisting of healthy volunteers. By adopting MALDI-TOF MS analysis, the serum samples obtained from the three groups of SLE patients were examined and compared with those from the control group; the categorized peptide fingerprint profile was established via the biological data collected from the samples., Results: The expression of protein with a m/z of 4207 Da increased significantly in SLE patients; the protein with a m/z of 2658 Da was expressed in all SLE patients; three proteins (with m/z of 1465, 5332, and 5900 Da respectively) were expressed in the SLE patients complicated with kidney injury and the protein with a m/z of 1943 Da was expressed in SLE patients with family history., Conclusion: A number of differential proteins were successfully detected and identified through MALDI-TOF MS detection and these proteins may be associated with the genetic basis of SLE and the complicating kidney injury.

Reactive oxygen species (ROS) play a critical role in renal ischemia-reperfusion injury (IRI). Intermedin (IMD) reportedly protected against myocardial IRI via its antioxidant effects; however, its protective role in renal IRI has not been investigated. We overexpressed IMD in rat kidneys and examined how the kidneys respond to renal IRI. Eukaryotic expression plasmid encoding the rat IMD gene or control empty vector was transfected into the left kidney using an ultrasound-microbubble-mediated delivery system. This method yielded high expression of IMD in kidney cells. Renal IRI was induced by clamping the left renal artery followed by reperfusion. In response to IRI, overexpression of IMD in the kidney significantly improved renal function and pathology compared with the kidney transfected with control plasmid. We investigated the mechanisms by which IMD protects against renal IRI. We examined renal superoxide dismutase (SOD) activity and malondialdehyde (MDA) content and found SOD activity was significantly increased, while MDA level was markedly decreased in kidneys transfected with IMD, suggesting ROS production and oxidative stress were reduced by IMD overexpression. We also measured myeloperoxidase (MPO) activity, tubular cell apoptosis, and the expression of intercellular adhesion molecule-1 (ICAM-1), P-selectin, and endothelin-1 (ET-1) in the kidney. Renal MPO activity and the expression of ICAM-1, P-selectin, and ET-1 stimulated by IRI were significantly inhibited by IMD overexpression. Moreover, IMD overexpression prevented kidney cells from apoptosis caused by IRI. Our results demonstrate that overexpression of IMD in the kidney protects against renal IRI, apparently by reducing oxidative stress, consequently suppressing inflammation and vasoconstrictor production and apoptosis.

Objective: To explore the therapeutic effects of leflunomide metabolite A771726 on high glucose-induced podocyte injury and understand its mechanism., Methods: The conditionally immortal human glomerular podocytes were divided into normal glucose group (NG), high glucose group (HG), mannitol group (MA), high glucose with SB202190 (a p38MAPK inhibitor) group and high glucose with active leflunomide metabolite A771726 group. The levels of p38MAPK and p-p38 protein were determined by Western blot. And the rate of podocyte apoptosis was evaluated by flow cytometry., Results: (1) Podocytes with high glucose could activate the p38MAPK signaling pathway. The p-p38 protein expression was significantly elevated. Little activation of the pathways was observed in Groups NG and MA. As compared to HG, the p-p38 protein expression was significantly lowered in SB202190 and A771726 groups. (2) Apoptosis increased in podocytes with high glucose after 24 h. The apoptotic rate of group HG was the most dramatic at 48 h (18.6 ± 0.7)%. It was significantly higher than those of Groups NG and MA. The group of high glucose with SB202190 and high glucose with active leflunomide metabolite A771726 could reduce the podocyte apoptosis by 26% and 17% respectively versus the HG group. And the difference was statistically significant., Conclusion: Leflunomide can inhibit high glucose-induced podocyte apoptosis. And this effect may be involved in its inhibition of activation of p38MAPK.

In the title compound, C(24)H(17)ClN(6), the dihedral angles between the triazolyl ring and its adjacent chlorobenzene and trisubstituted benzene rings are 90.6 (2) and 55.7 (3)°, respectively. The dihedral angle between the trisubstituted ring and the attached tolyl ring of the biphenyl unit is 45.9 (3)°. Intra- and intermolecular N-H⋯N hydrogen bonds are present.

Objective: To investigate the relationship between the polymorphism of microsomal triglyceride transfer protein gene (MTP-493G/T) and the level of serum lipids in Guangxi Heiyi Zhuang population., Methods: A total of 500 people of Heiyi Zhuang nationality, 272 males and 228 females, aged 7-83, were surveyed by cluster sampling. Blood pressure, height, weight, serum lipid and apolipoprotein levels were measured, and both body surface areas and body mass index were calculated. The genotype and allele of MTP-493G/T were determined by polymerase chain reaction combined with allele-specific oligonucleotide (PCR-ASO) hybridization method, and then confirmed by direct sequencing. The results were compared with those in 500 matched people of Han nationality who also live in that district., Results: (1) The frequencies of G and T allele were 0.74 and 0.26 in Heiyi Zhuang people, and 0.73 and 0.27 in Han people respectively. The frequencies of GG, GT and TT genotypes were 0.58 (289/500), 0.33 (166/500) and 0.09 (45/500) in Heiyi Zhuang, and 0.56 (281/500), 0.34 (172/500) and 0.09 (47/500) in Han people respectively. There were no significant differences of G/T allele and genotype frequencies between Heiyi Zhuang and Han people. (2) The levels of total cholesterol, triglyceride, low-density lipoprotein cholesterol, and apolipoprotein B in TT genotype were significantly higher than those in GT or GG genotype, but there were no significant differences of them between the GG and GT genotypes. The levels of high-density lipoprotein cholesterol and apolipoprotein A1 were not significantly different among the three genotypes., Conclusion: The frequencies of allele and genotype of MTP-493G/T are not significantly different between the Heiyi Zhuang and Han populations. But the levels of total cholesterol, triglyceride, low-density lipoprotein cholesterol, and apolipoprotein B in the TT genotype are significantly higher than those in the GT or GG genotype in both nationalities.