Paeoniflorin ameliorates AGEs-induced mesangial cell injury through inhibiting RAGE/mTOR/autophagy pathway.
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.
Key Laboratory of New Drug Delivery Systems of Chinese Materia Medica, Jiangsu Provincial Academy of Chinese Medicine, Jiangsu, Nanjing, 210028, PR China.
Juan
J
Chen
Department of Pharmacology of Chinese Materia Medica, China Pharmaceutical University, Nanjing, 210009, PR China.
Di
D
Zhao
Key Laboratory of New Drug Delivery Systems of Chinese Materia Medica, Jiangsu Provincial Academy of Chinese Medicine, Jiangsu, Nanjing, 210028, PR China.
Maomao
M
Zhu
Key Laboratory of New Drug Delivery Systems of Chinese Materia Medica, Jiangsu Provincial Academy of Chinese Medicine, Jiangsu, Nanjing, 210028, PR China.
Minghua
M
Zhang
Key Laboratory of New Drug Delivery Systems of Chinese Materia Medica, Jiangsu Provincial Academy of Chinese Medicine, Jiangsu, Nanjing, 210028, PR China.
Xuefeng
X
Hou
Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Jiangsu, Nanjing, 210028, PR China.
Wenbo
W
Ding
Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Jiangsu, Nanjing, 210028, PR China.
Shuai
S
Sun
Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Jiangsu, Nanjing, 210028, PR China.
Weiquan
W
Bu
Key Laboratory of New Drug Delivery Systems of Chinese Materia Medica, Jiangsu Provincial Academy of Chinese Medicine, Jiangsu, Nanjing, 210028, PR China; State Key Laboratory Breeding Base of Dao-di Herbs, China Academy of Chinese Medical Sciences, Beijng, 100700, PR China; Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Jiangsu, Nanjing, 210028, PR China. Electronic address: wenmoxiushi@163.com.
Liang
L
Feng
Department of Pharmacology of Chinese Materia Medica, China Pharmaceutical University, Nanjing, 210009, PR China. Electronic address: spma@cpu.edu.cn.
Shiping
S
Ma
Key Laboratory of New Drug Delivery Systems of Chinese Materia Medica, Jiangsu Provincial Academy of Chinese Medicine, Jiangsu, Nanjing, 210028, PR China; Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Jiangsu, Nanjing, 210028, PR China.
Xiaobin
X
Jia
D000818:Animals; D000071182:Autophagosomes / Q000187:drug effects / Q000378:metabolism; D001343:Autophagy / Q000187:drug effects*; D002460:Cell Line; D003928:Diabetic Nephropathies / Q000378:metabolism; D005960:Glucosides / Q000494:pharmacology*; D017127:Glycation End Products, Advanced / Q000378:metabolism*; D050527:Mesangial Cells / Q000187:drug effects* / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D039821:Monoterpenes / Q000494:pharmacology*; D051381:Rats; D000067759:Receptor for Advanced Glycation End Products / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects*; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism*; D015854:Up-Regulation / Q000187:drug effects; D006027:Glycosides
D005960:Glucosides; D017127:Glycation End Products, Advanced; D008869:Microtubule-Associated Proteins; D039821:Monoterpenes; D000067759:Receptor for Advanced Glycation End Products; D058570:TOR Serine-Threonine Kinases; C000593874:Ager protein, rat; C015423:peoniflorin; D006027:Glycosides; C546845:mTOR protein, rat
Advanced glycation end products; Autophagy; Mesangial cells; Paeoniflorin; RAGE/mTOR pathway