Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical University, Xuzhou 221004, Jiangsu, China.
Department of Obstetrics and Gynecology, Xuzhou Medical University Affiliated Hospital, Xuzhou 221006, Jiangsu, China.
Biochem Pharmacol. 2020 May;175:113849. doi: 10.1016/j.bcp.2020.113849. Epub 2020 Feb 12.
Gestational diabetes mellitus (GDM) is associated with an increased risk of progressing to type 2 DM and cardiovascular disease; however, the pathogenesis is still poorly understood. This study was to investigate roles of thrombin and its receptor protease-activated receptor 1 (PAR-1) and NLRP1 inflammasome in endothelial injury in GDM condition. Umbilical cord and plasma of GDM patients and high glucose (HG) cultured human umbilical vein endothelial cells (HUVECs) were used to examine the pathological changes of these pathways. Meanwhile, ameliorative effects and potential mechanisms of a natural product sarsasapogenin (Sar) were investigated in HUVECs. Thrombin/PAR-1 pathway, advanced glycation endproducts (AGEs) and their receptor (RAGE) axis, and the nucleotide-binding domain and leucine-rich repeat containing protein 1 (NLRP1) inflammasome were activated in GDM condition and HG-cultured HUVECs, accompanied by endothelial injury (decreased cell viability and increased lactate dehydrogenase release). Nevertheless, thrombin inhibition or PAR-1 antagonism caused decreases in AGEs formation and RAGE expression in HG-cultured HUVECs, while AGEs inhibition or RAGE antagonism declined PAR-1 expression not thrombin activity. Furthermore, thrombin inhibition or PAR-1 antagonism restrained NLRP1 inflammasome activation in HG-cultured HUVECs; meanwhile, NLRP1 expression and interleukin 18 levels were remarkably reduced in HG-cultured HUVECs after PAR-1 knockdown. Interestingly, Sar co-treatment could suppress thrombin/PAR-1 pathway, NLRP1 inflammasome, and AGEs/RAGE axis. Together, endothelial damages in GDM were likely due to enhanced interaction between AGEs/RAGE axis and thrombin/PAR-1 pathway, followed by NLRP1 inflammasome activation. Moreover, Sar may act as a protective agent against endothelial injury in chronic HG condition.
妊娠期糖尿病（GDM）与 2 型糖尿病和心血管疾病风险增加有关；然而，其发病机制仍知之甚少。本研究旨在探讨凝血酶及其受体蛋白酶激活受体 1（PAR-1）和 NLRP1 炎性体在 GDM 状态下内皮损伤中的作用。使用 GDM 患者的脐带和血浆以及高葡萄糖（HG）培养的人脐静脉内皮细胞（HUVEC）来检查这些途径的病理变化。同时，研究了天然产物薯蓣皂甙（Sar）在 HUVEC 中的改善作用及其潜在机制。在 GDM 条件和 HG 培养的 HUVEC 中，凝血酶/PAR-1 途径、晚期糖基化终产物（AGEs）及其受体（RAGE）轴和核苷酸结合域和富含亮氨酸重复序列的蛋白 1（NLRP1）炎性体被激活，伴随着内皮损伤（细胞活力降低和乳酸脱氢酶释放增加）。然而，在 HG 培养的 HUVEC 中，凝血酶抑制或 PAR-1 拮抗作用导致 AGEs 形成和 RAGE 表达减少，而 AGEs 抑制或 RAGE 拮抗作用降低了 PAR-1 表达而非凝血酶活性。此外，凝血酶抑制或 PAR-1 拮抗作用抑制了 HG 培养的 HUVEC 中 NLRP1 炎性体的激活；同时，PAR-1 敲低后，HG 培养的 HUVEC 中 NLRP1 表达和白细胞介素 18 水平显著降低。有趣的是，Sar 共同处理可以抑制凝血酶/PAR-1 途径、NLRP1 炎性体和 AGEs/RAGE 轴。总之，GDM 中的内皮损伤可能是由于 AGEs/RAGE 轴与凝血酶/PAR-1 途径之间的相互作用增强，随后激活 NLRP1 炎性体所致。此外，Sar 可能作为一种保护剂，防止慢性 HG 条件下的内皮损伤。
