Division of Nephrology, Beijing Anzhen Hospital, Capital Medical University, Beijing 100029, China.
J Diabetes Res. 2018 Nov 11;2018:1390418. doi: 10.1155/2018/1390418. eCollection 2018.
BACKGROUND/AIMS: It is known that chronic low-grade inflammation contributes to the initiation and development of both diabetes and diabetic nephropathy (DN), so we designed this study to investigate the role of P2X7R and NLRP3 inflammasome in DN pathogenesis and the antagonistic effects of artificially cultivated (ACOS).
A rat model of DN caused by high-fat-diet feeding and low-dose streptozotocin injection and a mouse podocyte injury model induced by high-glucose (HG) stimulation were established, and the intervention effects of ACOS on them were observed. The biological parameters of serum and urine and the pathological manifestations of kidney tissue were examined. The expression of mRNA and protein of P2X7R and NLRP3 inflammasome (NLRP3, ASC, and caspase-1) and downstream effectors (IL-1 and IL-18), as well as podocyte-associated molecules, was determined by real-time quantitative PCR and Western blot assay, respectively.
The DN rats showed to have developed insulin resistance, elevated fasting blood glucose, increased urinary protein excretion, and serum creatinine level as well as corresponding glomerular pathological alterations including podocyte damages. ACOS significantly antagonized the above changes. The experiments and both displayed that the mRNA and protein expression of P2X7R, NLRP3, ASC, caspase1 (procaspase-1 mRNA in the gene level and active caspase-1 subunit P10 in the protein level), IL-1, and IL-18 was significantly upregulated and the mRNA and protein expression of podocyte-associated molecules was significantly changed (downregulation of nephrin, podocin, and WT-1 expression and upregulation of desmin expression) indicating podocyte injury in the kidney tissue of DN rats and in the HG-stressed mouse podocytes, respectively. ACOS also significantly antagonized all the above changes.
Our research work suggests that P2X7R and NLRP3 inflammasome are involved in the pathogenesis of DN, and ACOS can effectively inhibit the high expression of P2X7R and the activation of NLRP3 inflammasome, which may contribute to the therapeutic effects of .
背景/目的：已知慢性低度炎症会导致糖尿病和糖尿病肾病（DN）的发生和发展，因此我们设计了这项研究，以探讨 P2X7R 和 NLRP3 炎性体在 DN 发病机制中的作用，以及人工栽培（ACOS）的拮抗作用。
建立高脂饮食喂养和小剂量链脲佐菌素注射诱导的大鼠 DN 模型和高糖（HG）刺激诱导的小鼠足细胞损伤模型，并观察 ACOS 对它们的干预作用。检测血清和尿液的生物学参数以及肾脏组织的病理表现。通过实时定量 PCR 和 Western blot 分别测定 P2X7R 和 NLRP3 炎性体（NLRP3、ASC 和 caspase-1）及其下游效应物（IL-1 和 IL-18）以及足细胞相关分子的 mRNA 和蛋白表达。
DN 大鼠表现出胰岛素抵抗、空腹血糖升高、尿蛋白排泄增加和血清肌酐水平升高，以及相应的肾小球病理改变，包括足细胞损伤。ACOS 显著拮抗了上述变化。实验 1 和实验 2 均显示，P2X7R、NLRP3、ASC、caspase1（基因水平的 procaspase-1 mRNA 和蛋白水平的活性 caspase-1 亚基 P10）、IL-1 和 IL-18 的 mRNA 和蛋白表达均显著上调，足细胞相关分子的 mRNA 和蛋白表达也发生显著变化（nephrin、podocin 和 WT-1 表达下调，desmin 表达上调），表明 DN 大鼠肾脏组织和 HG 刺激的小鼠足细胞中存在足细胞损伤。ACOS 也显著拮抗了所有上述变化。
我们的研究工作表明，P2X7R 和 NLRP3 炎性体参与了 DN 的发病机制，ACOS 能有效抑制 P2X7R 的高表达和 NLRP3 炎性体的激活，这可能有助于 ACOS 的治疗作用。
