目的探究脂肪含量和肥胖相关蛋白(fat mass and obesity-associated protein,FTO)和丝氨酸-苏氨酸激酶蛋白激酶D2(serine-threonine kinase protein kinase D2,PRKD2)在糖尿病肾病(diabetic kidney disease,DKD)进展中的调控作用和调节...目的探究脂肪含量和肥胖相关蛋白(fat mass and obesity-associated protein,FTO)和丝氨酸-苏氨酸激酶蛋白激酶D2(serine-threonine kinase protein kinase D2,PRKD2)在糖尿病肾病(diabetic kidney disease,DKD)进展中的调控作用和调节机制。方法采用35 mmol/L葡萄糖对足细胞(MPC5细胞)进行高糖刺激24h构建DKD体外模型。采用FTO过表达载体(pcDNA-FTO)和PRKD2过表达载体(pcDNA-PRKD2),或空载体(vector)转染高糖诱导的MPC5细胞。通过RT-qPCR检测FTO和PRKD2过表达效率;MeRIP检测PRKD2 mRNA的N6-甲基腺苷(N6-methyladenosine,m6A)修饰水平;ELISA检测Caspase-3活性、IL-6,TNF-α和单核细胞趋化蛋白-1(monocyte chemotactic protein-1,MCP-1)分泌量;流式细胞术分析细胞凋亡率;Western blot评估FTO和PRKD2蛋白水平,以及SIRT1/HIF-1α通路关键蛋白表达水平;Pearson分析FTO和PRKD2水平的相关性。结果与无高糖诱导对照组比较,高糖诱导的足细胞中FTO蛋白(0.51±0.04 vs 1.00±0.03)和PRKD2蛋白(0.45±0.03 vs 1.01±0.04)水平显著下调,差异具有统计学意义(t=13.17,16.76,均P<0.001)。高糖诱导的足细胞中FTO蛋白水平和PRKD2蛋白水平呈正相关(r2=0.7051,P<0.001)。与vector组相比,pcDNA-FTO组PRKD2 mRNA的m6A水平(0.56±0.09 vs1.01±0.13)降低,PRKD2 mRNA水平(3.16±0.14 vs 1.03±0.02)显著升高,差异具有统计学意义(t=51.37,11.82,均P<0.001)。与control组(IL-6:512.76±61.85 pg/ml,TNF-α:28.17±2.83 pg/ml,MCP-1:157.31±17.69 pg/ml)和vector组(IL-6:498.41±87.51 pg/ml,TNF-α:26.35±5.47 pg/ml,MCP-1:165.52±16.87 pg/ml)比较,pcDNA-PRKD2组IL-6(301.86±21.85 pg/ml),TNF-α(11.06±4.12 pg/ml),MCP-1分泌量(81.45±9.03pg/ml)显著减少,差异具有统计学意义(F=7.51,10.47,61.97,均P<0.01)。与control组(Caspase-3:689.65±79.5U/L,细胞凋亡率:22.31%±2.69%)和vector组(Caspase-3:715.91±113.58 U/L,细胞凋亡率:21.07%±3.28%)比较,pcDNA-PRKD2组Caspase-3活性(437.64±104.76 U/L)和细胞凋亡率(8.41%±3.15%)下降,差异具有统计学意义(F=2.35,79.13,均P<0.01)。与control组(SIRT1:1.01±0.05,HIF-1α:1.03±0.07)和vector组(SIRT1:0.97±0.05,HIF-1α:1.02±0.03)相比,pcDNA-PRKD2组SIRT1蛋白(3.51±0.15)水平升高,HIF-1α蛋白(0.37±0.07)水平降低,差异具有统计学意义(F=31.54,8.31,均P<0.01)。结论FTO介导m6A修饰的PRKD2通过SIRT1/HIF-1α通路抑制高糖诱导的足细胞炎症反应和细胞凋亡。展开更多
目的:分析25-羟基维生素D与DPN之间的关系。方法:选取2023年2月至2023年12月期间,在北华大学附属医院内分泌科住院的2型糖尿病患者99例,其中68例合并了周围神经病变(DPN),31例未合并周围神经病变(NDPN)。对两组之间的一般资料进行差异...目的:分析25-羟基维生素D与DPN之间的关系。方法:选取2023年2月至2023年12月期间,在北华大学附属医院内分泌科住院的2型糖尿病患者99例,其中68例合并了周围神经病变(DPN),31例未合并周围神经病变(NDPN)。对两组之间的一般资料进行差异性分析;logistic回归分析25-羟基维生素D与DPN之间的关系,并绘制受试者工作特征曲线(ROC)。结果:DPN组患者的年龄、糖尿病病程高于NDPN组患者(P P P P Objective: To analyze the relationship between 25-hydroxyvitamin D and DPN. Method: From February 2023 to December 2023, 99 patients with type 2 diabetes mellitus were hospitalized in the Department of Endocrinology, Affiliated Hospital of Beihua University, of which 68 had peripheral neuropathy (DPN) and 31 had no peripheral neuropathy (NDPN). Differences were analysed for general data between the two groups;logistic regression analysis was used to analyze the relationship between 25-hydroxyvitamin D and DPN, and the receiver operating characteristic curve (ROC) was plotted. Results: The age and duration of diabetes mellitus in the DPN group were higher than those in the NDPN group (P P P P < 0.05). Logistic regression multivariate analysis showed that the incidence of DPN increased by 19.1% for every 1 ng/ml decrease in 25-hydroxyvitamin D. The ROC curve area is 0.721. Conclusion: The level of 25-hydroxyvitamin D in patients with type 2 diabetes mellitus is significantly correlated with DPN. Evaluation of 25-hydroxyvitamin D may be a valuable part of the follow-up of patients with type 2 diabetes.展开更多
文摘目的:分析25-羟基维生素D与DPN之间的关系。方法:选取2023年2月至2023年12月期间,在北华大学附属医院内分泌科住院的2型糖尿病患者99例,其中68例合并了周围神经病变(DPN),31例未合并周围神经病变(NDPN)。对两组之间的一般资料进行差异性分析;logistic回归分析25-羟基维生素D与DPN之间的关系,并绘制受试者工作特征曲线(ROC)。结果:DPN组患者的年龄、糖尿病病程高于NDPN组患者(P P P P Objective: To analyze the relationship between 25-hydroxyvitamin D and DPN. Method: From February 2023 to December 2023, 99 patients with type 2 diabetes mellitus were hospitalized in the Department of Endocrinology, Affiliated Hospital of Beihua University, of which 68 had peripheral neuropathy (DPN) and 31 had no peripheral neuropathy (NDPN). Differences were analysed for general data between the two groups;logistic regression analysis was used to analyze the relationship between 25-hydroxyvitamin D and DPN, and the receiver operating characteristic curve (ROC) was plotted. Results: The age and duration of diabetes mellitus in the DPN group were higher than those in the NDPN group (P P P P < 0.05). Logistic regression multivariate analysis showed that the incidence of DPN increased by 19.1% for every 1 ng/ml decrease in 25-hydroxyvitamin D. The ROC curve area is 0.721. Conclusion: The level of 25-hydroxyvitamin D in patients with type 2 diabetes mellitus is significantly correlated with DPN. Evaluation of 25-hydroxyvitamin D may be a valuable part of the follow-up of patients with type 2 diabetes.
