Introduction and Objective: Diabetic Kidney Disease (DKD) is a leading cause of end-stage renal disease (ESRD). Extensive studies suggest that ferroptosis is a key factor in DKD progression. Our research aims to investigate whether ferroptosis-related factor ZFP36 can alleviate diabetic kidney injury. Methods: We extracted differentially expressed genes from the GEO database and conducted GO enrichment, GSEA, GSVA, plus machine learning techniques like RF, SVM-RFE, and Lasso Cox regression to identify ferroptosis-related genes. Expression differences of ZFP36 and its interacting proteins were validated in HK-2 cells and DKD mouse models. Additionally, ZFP36's interaction with traditional Chinese herbal components was assessed using AutoDock Tools to evaluate their impact on DKD ferroptosis. Results: We identified 9 genes significantly associated with both ferroptosis and DKD. Further machine learning marked ZFP36 as a central gene. In vivo experiments showed that compared to the control group, the expression of ZFP36 is reduced in the DKD model group. In vitro, overexpression of ZFP36 alleviates high-glucose-induced ferroptosis and reduce the formation of extracellular matrix components such as FN1 and α-SMA.ZFP36 interacts with Nrf2, reducing ROS and MDA levels, and increasing GSH levels, indicates that ZFP36 can alleviate oxidative stress. Molecular docking analysis shows that traditional Chinese herbs like Berberine and Jatrorrhizine positively regulating ferroptosis in DKD, exhibit high affinity for ZFP36, suggesting it mediates their renal protective effect. Conclusion: This study illuminates the emerging role of ZFP36 in ferroptosis in DKD, emphasizing its potential as a therapeutic target. This provides new insights for the development of targeted therapies for DKD. A. Li: None. M. Shi: None. W. Gu: None. Y. Chen: None. Y. Liu: None. J. Zhang: None. Y. Yuxuan: None. C. Huimin: None. J. Chen: None. H. Zhang: None. Huai'an Science and Technology Bureau (BE2023745), Jiangsu Provincial Health Commission (H2023137)

Background: The growing evidence has been verified that C1q/TNF-related protein 6 (CTRP6) and CTRP9, secreted by adipose tissue, can regulate glucose and lipid metabolism. However, the effect of CTRP6 and CTRP9 in gestational diabetes mellitus (GDM) is scarcely known. This study aimed to evaluate the potential clinical value of CTRP6 and CTRP9 in GDM. Methods: A total of 35 GDM subjects and 37 healthy controls, between 24 and 28 weeks of gestation, were included in the study. Diagnosis of GDM was made according to American Diabetes Association criteria. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of CTRP6 and CTRP9. Fasting insulin (FINS), IL-6, and TNF-α were detected by Luminex-xMAP technology. The main clinical characteristics, such as Anthropological data and metabolic parameters, were also obtained. Results: Circulating CTRP6, CTRP9, IL-6 and TNF-α were significantly elevated in GDM subjects compared with controls. Pearson correlation analysis showed that CTRP6 and CTRP9 were positively correlated with FPG (r = 0.280, P = 0.017 and r = 0.559, P < 0.001, respectively) and HOMA-IR (r = 0.250, P = 0.035 and r = 0.382, P = 0.001, respectively). Moreover, CTRP9 was also positively correlated with FINS (r = 0.253, P = 0.032), IL-6 (r = 0.283, P = 0.016) and TNF-α (r = 0.266, P = 0.024). The area under the receiver operating characteristic curve (AUC-ROC) indicated that the combination of CTRP6 and CTRP9 was more effective in predicting GDM than single indicators. Conclusion: Elevated CTRP6 and CTRP9 in GDM patients may be related to insulin resistance. And inflammatory factors may mediate the role of CTRP9 in insulin resistance. In addition, CTRP6 and CTRP9 may serve as candidate predictors in patients with early GDM. Disclosure Y.Liu: None. Y.Chen: None. W.Gu: None. L.Yin: None. H.Cao: None. M.Shi: None. Funding National Natural Science Foundation of China (81700723)

Objective: Damage to Schwann cells play a crucial role in diabetic peripheral neuropathy (DPN) . To observe whether HMGB1 inhibitor (Glycyrrhizic acid, GA) can prevent diabetic Schwann cells damage and its potential mechanisms. Methods: RSC96 cells were divided into 5 groups: NG group (5.6 mmol/L) and HG group (25.0 mmol/L) , HG+GA (1μM) group, HG+GA (10μM) , HG+GA (100μM) . Small interfering RNA was used to knock out HMGB1. CCK8 assay was used to detect the proliferation of RSC96 cells. The levels of inflammatory factors were determined by ELISA. The mRNA expression of NGF and neuritin-1 was detected by qRT-PCR. The expression levels of NSE, cleaved-caspase-3, RAGE, p38MAPK, ERK, JNK and NF-κBp65 in cells were measured by western blot. Results: GA at the concentration of μM for 24h had significant inhibitory effect on HMGB1 synthesis and secretion (P<0.05) . Schwann cell viability in high glucose was restored due to GA and siHMGB1 (P<0.05) . GA intervention and siHMGB1 potently prevented inflammatory substance generation (TNF-α, IL-1β, IL-6, MCP-1 and ICAM-1) , alleviated the neurotrophic factor reduction (NGF and neuritin-1) , and suppressed cellular apoptosis related protein activation (cleaved caspase-3) in Schwann cells exposed to high glucose ambience (P<0.05) . Moreover, we found that RAGE expression, p38MAPK phosphorylation and nuclear NF-κBp65 expression in high glucose-stimulated Schwann cells was reversed by GA or siRNA interference with HMGB1 (P<0.05) . Conclusions: GA and siHMGB1 can not only inhibit HMGB1 expression in Schwann cells exposed to high glucose, but also enhance cell viability. GA and siHMGB1 restored neurotrophic factors level and reduce cleaved caspase-3 expression in Schwann cell due to its anti-inflammatory ability, which may be related to the inhibition of RAGE/p38MAPK/nuclear NF-κBp65 pathway. HMGB1 may implicate in Schwann cells lesion in high glucose milieu and HMGB1 inhibition protects Schwann cells from high glucose induced cytotoxicity. Funding National Natural Science Foundation of China Grant Award (81700723)

Aim: To analyze the relationship between serum Elabela (ELA) levels and type 2 diabetic retinopathy (DR) . Methods: A total of 81 patients with type 2 diabetes were collected. According to the stages of diabetic retinopathy, the patients were divided into three groups: group 1: no diabetic retinopathy stage, group 2: non-proliferative diabetic retinopathy (NPDR) and group 3: proliferative diabetic retinopathy (PDR) , with 27 patients in each group. Serum ELA levels were detected by ELISA. Relevant clinical datas were recorded and analyzed. Results: There were no statistical significance in age, BMI, FPG, HbA1c, t-chol, TG, LDL-C, HDL-C among 3 groups (all P> 0.05) . The duration of diabetes in group 1 was significantly shorter than that in groups 2 and 3 (all P<0.05) , and there were significant differences in SBP, DBP, eGFR and Cre in groups 3 compared with those in groups 1 and 2 (P<0.05) . From group 1 to group 3, the levels of ELA decreased gradually, with statistical significance among the three groups (P<0.05) . Correlation analysis showed that serum ELA levels were negatively correlated with the course of diabetes, DR, SBP, Cre (P<0.05) , and positively correlated with eGFR (P<0.05) . Stepwise multiple linear regression analysis showed that the most relevant variables for ELA were age, BMI and DR (P=0.005; P = 0.000; P = 0.001) . According to ROC curve analysis, the sensitivity and specificity of ELA in the diagnosis of diabetic retinopathy were 59.3%, 83.3%, and the area under the curve was 0.753 (95%CI: 0.638, 0.869, P=0.000) . Conclusions: With the progressive of diabetic retinopathy, the level of serum ELA decreases gradually. ELA may be a potential clinical predictor and therapeutic target of diabetic retinopathy. Disclosure W.Gu: None. M.Shi: None. Y.Chen: None. Y.Liu: None. J.Song: None. H.Zhang: None. Funding National Natural Science Foundation of China Grant Award (81200595/81400807/81700723) , Six High-peak Talents Project of Jiangsu Province (WSN-101) , Research Project of Jiangsu 333 engineering (BRA2016232) and Research Project of Jiangsu Provincial Commission of Health and Family Planning (F201549/H201667) , and International Science and Technology Cooperation Project of Huaian (HAC201707) .

Objective: Damage to Schwann cells play a crucial role in diabetic peripheral neuropathy (DPN) . To observe whether HMGB1 inhibitor (Glycyrrhizic acid, GA) can prevent diabetic Schwann cells damage and its potential mechanisms. Methods: RSC96 cells were divided into 5 groups: NG group (5.6 mmol/L) and HG group (25.0 mmol/L) , HG+GA (1μM) group, HG+GA (10μM) , HG+GA (100μM) . Small interfering RNA was used to knock out HMGB1. CCK8 assay was used to detect the proliferation of RSC96 cells. The levels of inflammatory factors were determined by ELISA. The mRNA expression of NGF and neuritin-1 was detected by qRT-PCR. The expression levels of NSE, cleaved-caspase-3, RAGE, p38MAPK, ERK, JNK and NF-κBp65 in cells were measured by western blot. Results: GA at the concentration of μM for 24h had significant inhibitory effect on HMGB1 synthesis and secretion (P&lt;0.05) . Schwann cell viability in high glucose was restored due to GA and siHMGB1 (P&lt;0.05) . GA intervention and siHMGB1 potently prevented inflammatory substance generation (TNF-α, IL-1β, IL-6, MCP-1 and ICAM-1) , alleviated the neurotrophic factor reduction (NGF and neuritin-1) , and suppressed cellular apoptosis related protein activation (cleaved caspase-3) in Schwann cells exposed to high glucose ambience (P&lt;0.05) . Moreover, we found that RAGE expression, p38MAPK phosphorylation and nuclear NF-κBp65 expression in high glucose-stimulated Schwann cells was reversed by GA or siRNA interference with HMGB1 (P&lt;0.05) . Conclusions: GA and siHMGB1 can not only inhibit HMGB1 expression in Schwann cells exposed to high glucose, but also enhance cell viability. GA and siHMGB1 restored neurotrophic factors level and reduce cleaved caspase-3 expression in Schwann cell due to its anti-inflammatory ability, which may be related to the inhibition of RAGE/p38MAPK/nuclear NF-κBp65 pathway. HMGB1 may implicate in Schwann cells lesion in high glucose milieu and HMGB1 inhibition protects Schwann cells from high glucose induced cytotoxicity. Funding National Natural Science Foundation of China Grant Award (81700723)

Gestational diabetes mellitus (GDM) is a common disease in pregnant women, which mechanisms remain unclear. Apelin-APJ system contributes to the energy homeostasis in middle and terminal pregnancy. Elabela (ELA) is another endogenous ligand of apelin receptor (APJ), which is expressed highly in the placenta. Few studies have focused on the role of ELA in GDM and the association between ELA and inflammatory cytokines. In this study we mainly compare the concentrations of serum Elabela between 33 diabetes pregnant women (GDM group) with 33 healthy pregnant women (control group). The inflammatory biomarkers including IL-1β, IL-6 and TNF-α were measured and evaluated the relationship with ELA. There were significant differences in serum Elabela (P<0.05), fasting plasma glucose (FPG), one-hour glucose, two-hour glucose in the OGTT (P<0.001), HOMA-IR (P<0.001), cytokine IL-1β(P<0.05) between GDM group and control group. ELA had significant correlations with FPG (r=-0.254, P=0.039), fasting insulin (r=-0.258, P=0.036), HOMA-IR (r=-0.275, P=0.026), and IL-1β (r=-0.789, P=0.000). And stepwise multiple linear regression analysis presented IL-1β and FPG most significantly affected the ELA levels. This study showed that decreased ELA levels were significantly associated with increased glucose levels and insulin resistance in GDM, which might be mediated by inflammatory factors, such as IL-1β. Serum ELA levels might be a clinical biomarker for GDM. Disclosure Y. Chen: None. M. Shi: None. H. Zhang: None. Funding National Natural Science Foundation of China (81200595, 81400807)

Objective: ELABELA (ELA), an endogenous ligand for apelin receptor (APJ), is decreased in plasma of DKD patients. The effects of ELA on relieving diabetic renal lesion have not been reported. This study was performed to explore the protective impact and potential mechanisms of ELA on DKD via β-arrestins. Methods: Sixteen eight-week-old male db/db mice were randomly divided into diabetic nephropathy group (db/db group) and intervention group (db/db+ELA group) with administration of ELA (5mg ∙ kg-1 ∙ day-1) for 8 weeks. 8 age-matched male db/m mice were recognized as normal control group (db/m group). Both db/db and db/m group received equivalent normal saline injection for 8 weeks. Body weights and blood glucose levels were measured once a week. After 8 weeks, blood and urine samples were collected. Serum creatinine and urinary albumin to creatinine ration (UACR) were examined. The kidneys were taken after sacrificing mice. The morphology change of kidney tissues were assessed by HE and PAS staining. The expression of β-Arrestin1/2 was detected by immunohistochemistry. The levels of fibronectin (FN), collagen type IV (Col-IV) and β-arrestin1/2 in kidney tissues were examined by western blotting. Results: (1) With the intervention of ELA, UACR, serum creatinine and the ratio of kidney weight to body weight of db/db mice were significantly decreased (P<0.05). (2) Compared with db/m mice, db/db mice exhibited renal tubular epithelial cells edema, decreased ratio of non-lumen area to total area, thickened basement membrane and increased glycogen accumulation, however, those pathological abnormality were mitigated by ELA. (3) The levels of FN, Col-IV and β-Arrestin-1/2 in db/db mice were higher than db/m mice, while ELA treatment suppressed the expression of FN, COL-IV and β-Arrestin-1/2 (P<0.05). Conclusions: ELA may play a role in protection from diabetic kidney damage via down-regulating the expression of β-Arrestin-1/2 and inhibiting the secretion of extracellular matrix. Disclosure M. Shi: None. Y. Chen: None. H. Zhang: None. Funding National Natural Science Foundation of China (81700723)

目的 探讨T2DM患者血清Elabela(ELA)水平与DKD的关系.方法 选取2014年1月至2016年12月于我院内分泌科住院治疗的80例T2DM患者.ELISA法检测血清ELA水平,根据血清ELA水平分为高ELA组和低ELA组,检测两组患者HbA1c、TC、Scr、UACR等指标.结果 血清ELA总体水平为(7.18±3.17)ng/ml.与高ELA组比较,低ELA组UACR、Scr水平升高,eGFR降低(P<0.05);低ELA组DKD及DR患病率高于高ELA组(P<0.05).Pearson相关分析显示,血清ELA水平与eGFR、Ccr呈正相关(r=0.319、0.236,P<0.05),与UACR、Scr、SBP、DBP呈负相关(r=-0.561、-0.270、-0.249、-0.261,P<0.05).Logistic回归分析显示,UACR、BUN和LDL-C是血清ELA的影响因素.受试者工作特征曲线分析显示,ELA诊断DKD的敏感度为76.7％,特异度为80.0％,曲线下面积为0.803(95％CI 0.702～0.904,P=0.000).结论 T2DM合并白蛋白尿患者血清ELA较低,ELA可能是DKD潜在临床预测指标或治疗药物.

目的:探讨2型糖尿病患者血清促红素(EPO)和血清高迁移率蛋白B1(HMGB1)水平与尿蛋白的关系.方法:选取2型糖尿病患者80例,根据尿白蛋白/肌酐比值(UACR)分为正常蛋白尿组(UACR<30 mg/g)、微量蛋白尿组(UACR30～299 mg/g)、肾功能正常大量蛋白尿组(UACR≥300 mg/g)、肾功能不全大量蛋白尿组(UACR≥300 mg/g),每组各20例.检测所有患者血清EPO、HMGB1及相关临床生化指标水平.分析血清EPO、HMGB1之间的相关性及二者与UACR等其他临床生化指标水平的关系.结果:微量蛋白尿组、肾功能正常大量蛋白尿组和肾功能不全大量蛋白尿组糖尿病肾病(DKD)患者血清EPO浓度较正常蛋白尿组均明显降低,差异有统计学意义(P<0.05),且EPO浓度随着UACR增加而降低.EPO与尿素氮、血肌酐呈明显负相关,与肾小球滤过率(eGFR)呈正相关.肾功能正常大量蛋白尿组和肾功能不全大量蛋白尿组患者血清HMGB1浓度较正常蛋白尿组均显著增高,差异有统计学意义(P<0.01);HMGB1与BUN、Cr和UACR呈正相关,与eGFR呈负相关.EPO和HMGB1两者间呈负相关.结论:2型DKD患者血清EPO浓度降低、HMGB1水平升高,且与肾脏损害严重程度相关,血清EPO和HMGB1水平可为DKD的临床预测或治疗提供新思路.