Neferine inhibits the upregulation of CCL5 and CCR5 in vascular endothelial cells during chronic high glucose treatment.
We investigated whether the expressions of CCL5 and CCR5 participate in dysfunctional changes in human umbilical vein endothelial cells (HUVECs) induced by chronic high glucose treatment and examined whether neferine exerts its therapeutic effects by blocking the development of dysfunctional vascular endothelium. HUVECs were cultured with control or high concentrations of glucose in the absence or presence of neferine for 5 days. Nitric acid reductase method was used to detect the concentration of nitric oxide (NO) released into culture media. The level of intracellular reactive oxygen species (ROS) was measured by fluorescent DCFH-DA probe. The expressions of 84 genes related to endothelial cell biology were assessed by Human Endothelial Cell Biology RT(2) Profiler PCR Array. The expressions of the chemokine CCL5 and its receptor CCR5 were further determined by real-time RT-PCR and western blotting. PCR array indicated that CCL5 was the most significantly upregulated when HUVECs were exposed to chronic high glucose; the intracellular ROS level and the expressions of CCL5 and CCR5 at both mRNA and protein levels were significantly increased, whereas NO production was decreased simultaneously. The increased level of ROS and elevated expressions of CCL5 and CCR5 at high glucose were significantly inhibited by neferine; meanwhile the decreased NO production upon chronic high glucose treatment was relieved. An antioxidant (vitamin E) exerted similar beneficial effects. These data indicate that neferine can reduce the upregulation of CCL5 and CCR5 of vascular endothelium exposure to chronic high glucose and prevent or inhibit subsequent occurrence of inflammation in blood vessels possibly through antioxidation.
Department of Physiology, Basic Medical College of Nanchang University, 461 Bayi Road, Nanchang, Jiangxi, 330006, People's Republic of China.
Guilin
G
Li
Gaochun
G
Zhu
Yun
Y
Gao
Wen
W
Xiao
Hong
H
Xu
Shuangmei
S
Liu
Guihua
G
Tu
Haiying
H
Peng
Chaoran
C
Zheng
Shangdong
S
Liang
Guodong
G
Li
D000975:Antioxidants / Q000494:pharmacology; D044182:Benzylisoquinolines / Q000494:pharmacology*; D002470:Cell Survival; D002478:Cells, Cultured; D018946:Chemokine CCL5 / Q000096:biosynthesis*; D005947:Glucose / Q000008:administration & dosage / Q000494:pharmacology*; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007249:Inflammation / Q000188:drug therapy / Q000517:prevention & control; D009569:Nitric Oxide / Q000032:analysis / Q000378:metabolism; D010088:Oxidoreductases; D017382:Reactive Oxygen Species / Q000032:analysis; D019713:Receptors, CCR5 / Q000096:biosynthesis* / Q000187:drug effects; D015854:Up-Regulation / Q000187:drug effects; D014810:Vitamin E / Q000494:pharmacology
D000975:Antioxidants; D044182:Benzylisoquinolines; C483024:CCL5 protein, human; D018946:Chemokine CCL5; D017382:Reactive Oxygen Species; D019713:Receptors, CCR5; D014810:Vitamin E; C057222:neferine; D009569:Nitric Oxide; D010088:Oxidoreductases; C059847:nitric acid reductase; D005947:Glucose