title: Inhibition of Hypoxia-Induced Retinal Angiogenesis by Specnuezhenide, an Effective Constituent of Ligustrum lucidum Ait., through Suppression of the HIF-1α/VEGF Signaling Pathway.
abstract: Specnuezhenide (SPN), one of the main ingredients of Chinese medicine "Nü-zhen-zi", has anti-angiogenic and vision improvement effects. However, studies of its effect on retinal neovascularization are limited so far. In the present study, we established a vascular endothelial growth factor A (VEGFA) secretion model of human acute retinal pigment epithelial-19 (ARPE-19) cells by exposure of 150 μM CoCl₂ to the cells and determined the VEGFA concentrations, the mRNA expressions of VEGFA, hypoxia inducible factor-1α (HIF-1α) &amp; prolyl hydroxylases 2 (PHD-2), and the protein expressions of HIF-1α and PHD-2 after treatment of 3-(5'-hydroxymethyl-2'-furyl)-1-benzylindazole (YC-1, 1.0 μg/mL) or SPN (0.2, 1.0 and 5.0 μg/mL). Furthermore, rat pups with retinopathy were treated with SPN (5.0 and 10.0 mg/kg) in an 80% oxygen atmosphere and the retinal avascular areas were assessed through visualization using infusion of ADPase and H&amp;E stains. The results showed that SPN inhibited VEGFA secretion by ARPE-19 cells under hypoxia condition, down-regulated the mRNA expressions of VEGFA and PHD-2 slightly, and the protein expressions of VEGFA, HIF-1α and PHD-2 significantly in vitro. SPN also prevented hypoxia-induced retinal neovascularization in a rat model of oxygen-induced retinopathy in vivo. These results indicate that SPN ameliorates retinal neovascularization through inhibition of HIF-1α/VEGF signaling pathway. Therefore, SPN has the potential to be developed as an agent for the prevention and treatment of diabetic retinopathy.
authors.affiliation: Laboratory of Chinese Materia Medica, Department of Pharmacology, School of Pharmacy, Southwest Medical University, Luzhou 646000, Sichuan, China. jianmingwu@swmu.edu.cn.|Post-Doctoral Research Station, KangHong Pharmaceutical Group, Chengdu 610036, Sichuan, China. jianmingwu@swmu.edu.cn.|Post-Doctoral Mobile Station, West China School of Pharmacy, Sichuan University, Chengdu 610041, Sichuan, China. jianmingwu@swmu.edu.cn.
authors.forename: Jianming
authors.initials: J
authors.lastname: Wu
authors.affiliation: Post-Doctoral Research Station, KangHong Pharmaceutical Group, Chengdu 610036, Sichuan, China. kexiao@cnkh.com.
authors.forename: Xiao
authors.initials: X
authors.lastname: Ke
authors.affiliation: Post-Doctoral Research Station, KangHong Pharmaceutical Group, Chengdu 610036, Sichuan, China. fu_wei99@163.com.
authors.forename: Wei
authors.initials: W
authors.lastname: Fu
authors.affiliation: Post-Doctoral Research Station, KangHong Pharmaceutical Group, Chengdu 610036, Sichuan, China. 1483687108@qq.com.
authors.forename: Xiaoping
authors.initials: X
authors.lastname: Gao
authors.affiliation: Post-Doctoral Research Station, KangHong Pharmaceutical Group, Chengdu 610036, Sichuan, China. arken20@163.com.
authors.forename: Hongcheng
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Post-Doctoral Research Station, KangHong Pharmaceutical Group, Chengdu 610036, Sichuan, China. dy_ts110@163.com.
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.affiliation: Post-Doctoral Research Station, KangHong Pharmaceutical Group, Chengdu 610036, Sichuan, China. manapppp@163.com.
authors.forename: Na
authors.initials: N
authors.lastname: Ma
authors.affiliation: Post-Doctoral Research Station, KangHong Pharmaceutical Group, Chengdu 610036, Sichuan, China. zhaomanqian@cnkh.com.
authors.forename: Manxi
authors.initials: M
authors.lastname: Zhao
authors.affiliation: Post-Doctoral Research Station, KangHong Pharmaceutical Group, Chengdu 610036, Sichuan, China. hxf@cnkh.com.
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Hao
authors.affiliation: Post-Doctoral Mobile Station, West China School of Pharmacy, Sichuan University, Chengdu 610041, Sichuan, China. zrzzl@vip.sina.com.
authors.forename: Zhirong
authors.initials: Z
authors.lastname: Zhang
mesh_terms: D020533:Angiogenesis Inhibitors / Q000302:isolation & purification / Q000494:pharmacology*; D000818:Animals; D015687:Cell Hypoxia; D002460:Cell Line; D003035:Cobalt / Q000494:pharmacology; D003930:Diabetic Retinopathy / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D004195:Disease Models, Animal; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation; D005960:Glucosides / Q000302:isolation & purification / Q000494:pharmacology*; D006801:Humans; D000860:Hypoxia / Q000150:complications / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases / Q000235:genetics / Q000378:metabolism; D007191:Indazoles / Q000494:pharmacology; D031659:Ligustrum / Q000737:chemistry*; D010936:Plant Extracts / Q000737:chemistry; D011714:Pyrans / Q000302:isolation & purification / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015861:Retinal Neovascularization / Q000188:drug therapy* / Q000209:etiology / Q000235:genetics / Q000473:pathology; D055213:Retinal Pigment Epithelium / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D015398:Signal Transduction; D042461:Vascular Endothelial Growth Factor A / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism
chemical_list: D020533:Angiogenesis Inhibitors; D005960:Glucosides; C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D007191:Indazoles; D010936:Plant Extracts; D011714:Pyrans; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; C090937:3-(5'-hydroxymethyl-2'-furyl)-1-benzylindazole; C083178:nuezhenide; D003035:Cobalt; C490190:EGLN1 protein, human; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases; C018021:cobaltous chloride
keywords: angiogenesis; hypoxia-inducible factor-1; oxygen-induced retinopathy; specnuezhenide; vascular endothelial growth factor