CD38 Deficiency Protects Mouse Retinal Ganglion Cells Through Activating the NAD+/Sirt1 Pathway in Ischemia-Reperfusion and Optic Nerve Crush Models.

Purpose: The purpose of this study was to investigate the protective effect of CD38 deletion on retinal ganglion cells (RGCs) in a mouse retinal ischemia/reperfusion (I/R) model and an optic nerve crush (ONC) model, and to elucidate the underlying molecular mechanisms. Methods: Retinal I/R and ONC models were constructed in mice. PCR was used to identify the deletion of CD38 gene in mice, hematoxylin and eosin (H&E) staining was used to evaluate the changes in retinal morphology, and electroretinogram (ERG) was used to evaluate the changes in retinal function. The survival of RGCs and activation of retinal macroglia were evaluated by immunofluorescence staining. The expression of Sirt1, CD38, Ac-p65, Ac-p53, TNF-α, IL-1ß, and Caspase3 proteins in the retina was further evaluated by protein imprinting. Results: In retinal I/R and ONC models, CD38 deficiency reduced the loss of RGCs and activation of macroglia and protected the retinal function. CD38 deficiency increased the concentration of NAD+, reduced the degree of acetylation of NF-κB p65 and p53, and reduced expression of the downstream inflammatory cytokines TNFα, IL-1ß, and apoptotic protein Caspase3 in the retina in the ONC model. Intraperitoneal injection of the Sirt1 inhibitor EX-527 partially counteracted the effects of CD38 deficiency, suggesting that CD38 deficiency acts at least in part through the NAD+/Sirt1 pathway. Conclusions: CD38 plays an important role in the pathogenesis of retinal I/R and ONC injury. CD38 deletion protects RGCs by attenuating inflammatory responses and apoptosis through the NAD+/Sirt1 pathway.

Retinal microvasculature features in patients with migraine: a systematic review and meta-analysis.

Background: Migraine is a central nervous system disorder involving neuronal and vascular factors. The brain has a close anatomical relationship with retinal vessels and similar regulatory processes, and the retinal vascular system is the only in vivo vessel that can be directly visualized, while optical coherence tomography angiography (OCTA) is an advanced retinal vascular imaging technique. In this study, OCTA was used to study the retinal vascular density (VD) and foveal avascular zone (FAZ) in migraine patients, which provided a theoretical basis for its use as a candidate for rapid and non-invasive diagnosis of migraine. Methods: Published studies comparing retinal microvascular profiles between migraine patients and healthy controls were obtained by a comprehensive search of electronic databases. Nine studies were finally included, including 775 eyes (migraine group: 444 eyes, control group: 331 eyes). Pooled effect sizes were presented as standardized mean differences (SMDs) and 95% confidence intervals (CIs). Statistical analysis was performed using Review Manager software (version 5.30). Results: The combined results revealed that the superficial and deep macular whole enface VD (MWEVD) (superficial VD: SMD = -0.30, P = 0.0001; deep VD: SMD = -0.61, P = 0.02), superficial foveal VD (FVD) (SMD = -0.42, P = 0.03), deep parafoveal VD (PFVD) (SMD = -0.31, P = 0.002), and peripapillary VD (PVD) (SMD = -0.49, P = 0.002) were significantly reduced in migraine patients compared with healthy people. However, there was a significant increase in the area of the FAZ in migraine patients (SMD = 0.56, P < 0.0001). Conclusion: Migraine patients are prone to retinal microcirculation disorders, such as decreased blood vessel density and increased avascular area in the fovea. This provides a theoretical basis for OCTA as a candidate for rapid, non-invasive diagnosis of migraine.

Long-term glucocorticoid exposure persistently impairs CD4+ T cell biology by epigenetically modulating the mTORC1 pathway.

Conventional glucocorticoid (GC) treatment has a long-term influence on T-cell immunity, resulting in an increased risk of opportunistic infection after drug withdrawal. The underlying mechanisms remain ambiguous. This study demonstrated that long-term GC treatment induced persistent lymphopenia in patients with primary glomerular disease. GCs continuously suppressed the proportion of CD4+ T cells even after the daily dose was tapered down to the physiologic equivalences, leading to a significant decline of the CD4/CD8 ratio. Meanwhile, GCs impaired CD4+ T cell biology, leading to enhanced apoptotic cell death, reduced proliferative capacity, downregulated pro-inflammatory genes, and upregulated immunoregulatory genes. Specifically, GCs altered FOXP3 expression pattern in CD4+ T cells and favored their acquisition of an active T regulatory (Treg) cell phenotype with enhanced IL-10 production upon stimulation. Mechanistically, GCs tampered with the transcriptional regulation of mechanistic target of rapamycin complex 1 (mTORC1) pathway, resulting in an inhibitory impact on the signaling activity. Targeting mTORC1 signaling by siRNAs could sufficiently modify the viability of GC-exposed CD4+ T cells. By high-throughput sequencing of genome-wide DNA methylation and mRNA, we further uncovered a causal relationship between the altered DNA methylation level and transcription activity in a subset of mTORC1 pathway genes in long-term GC exposure. Taken together, this study reveals a novel regulation of mTORC1 signaling, which might dominate the long-term influence of GC on CD4+ T cell biology in a dose-independent manner.

Autophagy in Rat Müller Glial Cells Is Modulated by the Sirtuin 4/AMPK/mTOR Pathway and Induces Apoptosis under Oxidative Stress.

Müller glial cells (MGCs) are a group of glial cells in the retina that provide essential support to retinal neurons; however, the understanding of MGC apoptosis and autophagy remains limited. This study was aimed at investigating the role of autophagy in MGCs under normal and oxidative conditions, and identifying the underlying mechanisms. In addition, the sirtuin 4 (SIRT4)-mediated signaling pathway was observed to regulate the autophagic process in MGCs. To assess the effect of autophagy on MGC mitochondrial function and survival, we treated rMC-1 cells-rat-derived Müller glial cells-with rapamycin and 3-methyladenine (3-MA), and found that MGC death was not induced by such treatment, while autophagic dysfunction could increase MGC apoptosis under oxidative stress, as reflected by the expression level of cleaved caspase 3 and PI staining. In addition, the downregulation of autophagy by 3-MA could influence the morphology of the mitochondrial network structure, the mitochondrial membrane potential, and generation of reactive oxygen species (ROS) under oxidative stress. Moreover, SIRT4 depletion enhanced autophagosome formation, as verified by an increase in the LC3 II/I ratio and a decrease in the expression of SQSTM1/p62, and vice versa. The inhibition of AMPK phosphorylation by compound C could reverse these changes in LC3 II/I and SQSTM1/p62 caused by SIRT4 knockdown. Our research concludes that MGCs can endure autophagic dysfunction in the absence of oxidative stress, while the downregulation of autophagy can cause MGCs to become more sensitized to oxidative stress. Simultaneous exposure to oxidative stress and autophagic dysfunction in MGCs can result in a pronounced impairment of cell survival. Mechanically, SIRT4 depletion can activate the autophagic process in MGCs by regulating the AMPK-mTOR signaling pathway.

The predictive value of circular RNAs in the diagnosis, prognosis and clinicopathological features of thyroid cancer: A systematic review and meta-analysis.

OBJECTIVE: An increasing number of studies have reported circular RNAs (circRNAs) as new potential diagnostic and prognostic biomarkers for thyroid cancer. However, the overall predictive value of circRNAs for thyroid cancer remains unclear. Therefore, we performed a comprehensive meta-analysis to investigate the predictive value of circRNAs in the diagnosis, prognosis and clinicopathological features of thyroid cancer (TC). METHODS: PubMed, Embase, Cochrane library and Web of science were searched systematically. The search time limit was from the establishment of the database to October 2021. We used the subject words combined with free words for the search. Furthermore, we used Review Manager 5.3 and Meta-Disc1.4 for relevant data analysis, and Stata 13.1 for sensitivity analysis and publication bias test. The combined sensitivity and specificity were used to evaluate the diagnostic value of circRNAs. The hazard ratio (HR) and its 95% confidence interval (95%CI) were used for statistical analysis of the prognosis of TC. The clinical characteristics were analyzed with Odds Ratio (OR) and its 95% CI. P = 0.05 for all analysis, Q test and I2 analysis were used for heterogeneity evaluation, I2 ≤ 50% and Q test P > 0.1, indicated the lack of heterogeneity. RESULTS: A total of 26 eligible studies including 18 on clinicopathological studies, 7 on prognostic and 6 diagnostic studies were enrolled in our meta-analysis. A total of 1911 patients with TC were included. For the clinicopathological and prognostic value, the results showed that a high expression of circRNAs was correlated with poor clinical parameters, such as the tumor size [OR = 2.79, 95%CI (1.88, 4.13)], extra-glandular metastasis (ETE) [OR = 0.74, 95%CI (0.45, 1.53),P = 0.24], TNM staging [OR = 3.82, 95%CI (2.86, 5.10)], lymph node metastasis (LNM) [OR = 3.88, 95% (2.90, 5.19)], tumor focality [OR = 31.87, 95%CI (1.05, 3.31)] and poor overall survival (OS)[ HR = 1.97, 95%CI (1.37, 2.83)]. For the diagnostic value, the sensitivity, specificity and Area Under Curve (AUC) of the combined group were 0.83 [95%CI (0.77-0.87)], 0.77 [95%CI (0.71-0.82)] and 0.91 respectively. CONCLUSIONS：: CircRNAs have a potential diagnostic value for TC. Meanwhile, an elevated expression of circRNAs has been linked to a poor OS in TC patients, and it also associates with poor prognosis of thyroid cancer patients. The expression of circRNAs is also correlated with certain clinical characteristics, suggesting that it may have important clinical significance.

CD38 deficiency protects the retina from ischaemia/reperfusion injury partly via suppression of TLR4/MyD88/NF-κB signalling.

PURPOSE: This study aimed to explore cellular localisation of CD38 in the retina and evaluate the role and potential mechanism of CD38 deficiency in retinal ischaemia/reperfusion (I/R) injury. METHODS: Six-to eight-week-old male CD38 knockout (KO) and wild-type mice in C57BL/6 background were used. Immunostaining was performed to determine the cellular localisation of CD38 in the retina. Haematoxylin and eosin staining and immunostaining of Brn3a were used to evaluate the retinal I/R injury. Western blotting was performed to detect toll-like receptor 4 (TLR4), myeloid differentiation primary response 88 (MyD88), p-p65, ionised calcium-binding adapter molecule 1, Sirtuin1 (Sirt1), Ac-p65, and pro-inflammatory cytokines protein expression. RESULTS: CD38 was highly expressed in mouse retinal microglia and astrocytes/Müller cells. CD38 deficiency reduced I/R-induced retinal damage and retinal ganglion cell death. Following retinal I/R injury, TLR4, MyD88, nuclear factor-κB p-p65 (NF-κB p-p65), pro-inflammatory cytokines and CD38 protein levels were also upregulated. After I/R injury, retinal inflammation factors IL-1ß, IL-6, and TNF-α mRNA and protein levels were increased. IL-1ß, IL-6, and TNF-α were reduced in CD38 KO mice after I/R injury. Retinal I/R injury induced the activation of microglia, but this effect was also suppressed by KO of CD38. Additionally, retinal I/R induced a significant increase in Ac-p65 protein levels and decrease in Sirt1 protein levels, while this effect was greatly attenuated by KO of CD38. CONCLUSION: CD38 deficiency protects the retina from I/R injury by suppressing microglial activation partly via activating Sirt1-mediated suppression of TLR4/MyD88/NF-κB signalling.