Key Laboratory of New Drug Delivery Systems of Chinese Materia Medica, Jiangsu Provincial Academy of Chinese Medicine, Jiangsu, Nanjing, 210028, PR China.
Department of Pharmacology of Chinese Materia Medica, China Pharmaceutical University, Nanjing, 210009, PR China.
Biomed Pharmacother. 2017 May;89:1362-1369. doi: 10.1016/j.biopha.2017.03.016. Epub 2017 Mar 18.
Glomerular mesangial cell plays a vital role in diabetic nephropathy (DN). Recent research has demonstrated that autophagy involved in the development of DN. Paeoniflorin (PF), a monoterpene glucoside, has been proved to attenuate advanced glycation end products (AGEs)-induced mesangial cell injury. However, the regulatory mechanism of PF on autophagy in mesangial cell remains unclear. The aim of this study was to explore the effect of PF on autophagy in AGEs-induced mesangial cell dysfunction. In this study, the leakage of the lactic dehydrogenase (LDH) into the extracellular medium was measured by LDH kit. Transmission electron microscopy (TEM) and mRFP-GFP-microtubule-associated protein light chain 3 (LC3) transfection were performed to observe the formation of autophagy in AGEs-induced mesangial cell. The RAGE/mTOR/autophagy pathway was analyzed by western blotting and small-interfering RNA transfection. Our results showed that the expression of LC3II, p62 were changed in a time-dependent manner in AGEs-stimulated mesangial cell. While PF could decrease the expression of LC3II/LC3I and reduce the number of autophagosomes. Knockdown of Atg5 promoted the protective effect of PF on AGEs-induced HBZY-1 injury. Furthermore, we found PF inhibited autophagy at least partly through inhibiting RAGE and upregulating the level of p-mTOR to against AGEs-induced mesangial cell dysfunction. Thus, PF could be a potential agent for the treatment of DN.
肾小球系膜细胞在糖尿病肾病 (DN) 中起着至关重要的作用。最近的研究表明，自噬参与了 DN 的发展。芍药苷 (PF) 是一种单萜糖苷，已被证明可以减轻晚期糖基化终产物 (AGEs) 诱导的系膜细胞损伤。然而，PF 对系膜细胞自噬的调节机制尚不清楚。本研究旨在探讨 PF 对 AGEs 诱导的系膜细胞功能障碍中自噬的影响。在这项研究中，通过 LDH 试剂盒测量乳酸脱氢酶 (LDH) 向细胞外培养基中的漏出。通过透射电子显微镜 (TEM) 和 mRFP-GFP-微管相关蛋白轻链 3 (LC3) 转染观察 AGEs 诱导的系膜细胞中自噬的形成。通过 Western blot 和小干扰 RNA 转染分析 RAGE/mTOR/自噬途径。我们的结果表明，在 AGEs 刺激的系膜细胞中，LC3II 和 p62 的表达呈时间依赖性变化。而 PF 可以降低 LC3II/LC3I 的表达并减少自噬体的数量。Atg5 的敲低促进了 PF 对 AGEs 诱导的 HBZY-1 损伤的保护作用。此外，我们发现 PF 通过抑制 RAGE 和上调 p-mTOR 水平至少部分抑制自噬，从而对抗 AGEs 诱导的系膜细胞功能障碍。因此，PF 可能是治疗 DN 的潜在药物。
