Division of Nephrology, Nanfang Hospital, Southern Medical University, Research Institute of Nephrology Guangdong Province, Key Laboratory for Organ Failure Research, Ministry of Education, Guangzhou, Guangdong 510515, China.
Chin Med J (Engl). 2013 Apr;126(7):1230-5.
Diabetic nephropathy (DN) is the leading cause of end-stage renal disease. Various treatment regimens and combinations of therapies provide only partial renoprotection. Therefore new approaches are needed to retard the progression of DN. The aim of the present study was to evaluate the role of a novel spiroalkaloid from Acorus tatarinowii named acortatarin A (AcorA) in inhibiting high glucose-induced extracellular matrix accumulation in mesangial cells (MCs).
The cytotoxity of AcorA on MCs was examined by 3-(4,5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide (MTT) assay. The expression of fibronectin and collagen IV was examined by real time PCR and western blotting. The expression of p22(phox) and p47(phox) was detected by western blot. The interaction between p22(phox) and p47(phox) was examined by co-immunoprecipitation. The phosphorylation of p47(phox) was examined by immunoprecipitation. The phosphorylation of protein kinase C (PKC) α, PKCβ, phospholiase C gamma (PLCγ1), and the p85 subunit of PI3K was determined by Western blotting.
AcorA significantly inhibited high glucose-induced activation of NADPH oxidase, a ROS-generating enzyme, by increasing phosphorylation of p47(phox) and enhancing interaction between p22(phox) and p47(phox). Preincubation of AcorA with MCs inhibited high glucose-induced collagen IV and fibronectin production in a dose-dependent manner. Moreover, AcorA attenuated high glucose enhanced phosphorylation of PKCα, PKCβ, PLCγ1, and the p85 subunit of PI3K.
AcorA inhibits high glucose-induced extracellular matrix production via blocking NADPH oxidase activation.
糖尿病肾病（DN）是终末期肾病的主要原因。各种治疗方案和治疗组合仅提供部分肾脏保护。因此，需要新的方法来延缓 DN 的进展。本研究旨在评估从菖蒲中分离得到的一种新型螺环生物碱命名为菖蒲 A（AcorA）在抑制高糖诱导的系膜细胞（MC）细胞外基质积聚中的作用。
通过 3-(4,5-二甲基噻唑-2-基)-2.5-二苯基四氮唑溴盐（MTT）测定法检测 AcorA 对 MCs 的细胞毒性。实时 PCR 和 Western blot 检测纤维连接蛋白和胶原 IV 的表达。Western blot 检测 p22(phox)和 p47(phox)的表达。通过共免疫沉淀检测 p22(phox)和 p47(phox)之间的相互作用。通过免疫沉淀检测 p47(phox)的磷酸化。Western blot 检测蛋白激酶 C（PKC）α、PKCβ、磷酸脂酶 Cγ（PLCγ1）和 PI3K 的 p85 亚基的磷酸化。
AcorA 通过增加 p47(phox)的磷酸化和增强 p22(phox)和 p47(phox)之间的相互作用，显著抑制了高糖诱导的 NADPH 氧化酶（一种 ROS 生成酶）的激活。AcorA 预处理 MCs 可剂量依赖性抑制高糖诱导的胶原 IV 和纤维连接蛋白产生。此外，AcorA 减弱了高糖增强的 PKCα、PKCβ、PLCγ1 和 PI3K 的 p85 亚基的磷酸化。
AcorA 通过阻断 NADPH 氧化酶的激活抑制高糖诱导的细胞外基质产生。
