金合欢素调节Hippo信号通路对糖尿病视网膜病变大鼠血管生成的影响

doi:10.11958/20230889

天津医药 ›› 2024, Vol. 52 ›› Issue (6): 578-583.doi: 10.11958/20230889

金合欢素调节Hippo信号通路对糖尿病视网膜病变大鼠血管生成的影响

陈芋洁(), 黄霞, 邓铂林, 贾文文()

四川省医学科学院·四川省人民医院眼科（邮编610000）

收稿日期:2023-06-12 修回日期:2023-08-30 出版日期:2024-06-15 发布日期:2024-06-06
通讯作者: ^△E-mail：hailunx60@163.com
作者简介:陈芋洁（1993），女，主要从事眼科方面研究。E-mail：zznijnmn@163.com
基金资助:
四川省科技计划项目(23ZDYF1942)

Effects of acacetin on angiogenesis in diabetes retinopathy rats by regulating Hippo signaling pathway

CHEN Yujie(), HUANG Xia, DENG Bolin, JIA Wenwen()

Department of Ophthalmology, Sichuan Academy of Medical Sciences, Sichuan Provincial People's Hospital, Chengdu 610000, China

Received:2023-06-12 Revised:2023-08-30 Published:2024-06-15 Online:2024-06-06
Contact: ^△E-mail: hailunx60@163.com

陈芋洁, 黄霞, 邓铂林, 贾文文. 金合欢素调节Hippo信号通路对糖尿病视网膜病变大鼠血管生成的影响[J]. 天津医药, 2024, 52(6): 578-583.

CHEN Yujie, HUANG Xia, DENG Bolin, JIA Wenwen. Effects of acacetin on angiogenesis in diabetes retinopathy rats by regulating Hippo signaling pathway[J]. Tianjin Medical Journal, 2024, 52(6): 578-583.

摘要/Abstract

摘要：

目的探究金合欢素（Aca）调节Hippo信号通路对糖尿病视网膜病变（DR）大鼠血管生成的影响。方法将60只SD大鼠随机分为对照组、DR组、Aca低剂量组（10 mg/kg）、Aca中剂量组（20 mg/kg）、Aca高剂量组（30 mg/kg）、Hippo-yes相关蛋白（YAP）通路抑制剂Verteporfin组（Aca高剂量30 mg/kg+Verteporfin 0.8 pmol/kg），每组10只。除对照组外，采用链脲佐菌素和高脂饲料喂养构建DR模型，检测大鼠体质量和空腹血糖（FBG）水平，荧光素血管造影（FFA）观察视网膜血管新生和荧光素渗漏，酶联免疫吸附试验检测血清血管内皮生长因子（VEGF）、促血管生成素-2（Ang-2）水平，苏木素伊红染色观察视网膜组织病理形态变化，Western blot法检测VEGF、缺氧诱导因子1α（HIF-1α）、血管细胞黏附分子-1（VCAM-1）及Hippo信号通路蛋白表达。结果与对照组比较，DR组大鼠视网膜细胞排列混乱、松散，新生血管形成，大量荧光素渗漏，体质量、大肿瘤抑制激酶2（LATS2）、磷酸化YAP（p-YAP）表达降低，FBG、VEGF、Ang-2、HIF-1α、VCAM-1、YAP、转录调节因子（TAZ）、TEA结构域家族成员1（TEAD1）表达增加（P<0.05）；与DR组比较，Aca低、中、高剂量组和Verteporfin组大鼠视网膜细胞排列整齐，新生血管形成和荧光素渗漏减少，体质量、LATS2、p-YAP表达增加，FBG、VEGF、Ang-2、HIF-1α、VCAM-1、YAP、TAZ、TEAD1表达降低（P<0.05），高剂量组效果更明显，但Verteporfin组与Aca高剂量组对DR大鼠各指标的作用效果差异无统计学意义。结论 Aca能抑制DR大鼠血管生成，改善视网膜病理损伤，其作用机制可能与调节Hippo信号通路有关。

关键词: 糖尿病视网膜病变, 视网膜新生血管化, 金合欢素, Hippo信号通路

Abstract:

Objective To investigate the impact of acacetin (Aca) on angiogenesis in diabetes retinopathy (DR) rats by regulating Hippo signaling pathway. Methods Sixty SD rats were grouped into the control group, the DR group, the Aca low dose group (10 mg/kg), the Aca medium dose group (20 mg/kg), the Aca high dose group (30 mg/kg) and Hippo-YAP signaling pathway inhibitor Verteporfin group (Aca high dose 30 mg/kg+Verteporfin 0.8 pmol/kg), with 10 rats in each group. Except the control group, streptozotocin and high-fat feed were used to construct the DR model. Body weight and fasting blood glucose (FBG) levels of rats were measured. Fluorescein angiography (FFA) was applied to observe retinal angiogenesis and fluorescein leakage. Enzyme linked immunosorbent assay was applied to detect serum levels of vascular endothelial growth factor (VEGF) and angiopoietin-2 (Ang-2). Pathological changes of retinal tissue were observed by hematoxylin-eosin staining. Western blot assay was applied to detect expression levels of VEGF, hypoxia-inducible factor 1 alpha (HIF-1α), vascular cell adhesion molecule-1 (VCAM-1) and Hippo signaling pathway proteins. Results Compared with the control group, retinal cells of rats in the DR group were arranged in a disordered and loose manner, with neovascularization and a large amount of fluorescein leakage, and body weight, the expression of large tumor suppressor homolog 2 (LATS2), p-Yes-associated protein (YAP) were reduced. FBG and expression levels of VEGF, Ang-2, HIF-1α, VCAM-1, YAP, transcription activator with PDZ binding motif (TAZ), TEA domain family member 1 (TEAD1) were increased (P<0.05). Compared with the DR group, retina cells of rats in the Aca low, medium and high-dose groups and the Verteporfin group were arranged neatly, with reduced neovascularization and fluorescence leakage, body weight and the expression levels of LATS2 and p-YAP were increased, and FBG, expression levels of VEGF, Ang-2, HIF-1α, VCAM-1, YAP, TAZ and TEAD1 were reduced (P<0.05). The effect was more obvious in the Aca high dose group. However, there was no significant difference in each indicator between the Verteporfin group and the Aca high dose group. Conclusion Aca can inhibit angiogenesis and improve retinal pathological damage in DR rats, and its mechanism of action may be related to regulating the Hippo signaling pathway.

Key words: diabetic retinopathy, retinal neovascularization, acacin, Hippo signaling pathway

中图分类号:

R774.1

图/表 8

参考文献 24

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组别	体质量/g	FBG/（mmol/L）
对照组	432.68±45.09	5.27±0.62
DR组	285.93±29.65^a	23.59±2.61^a
Aca低剂量组	358.76±36.92^b	15.62±1.61^b
Aca中剂量组	382.05±39.02^bc	10.07±1.14^bc
Aca高剂量组	417.25±42.83^bcd	6.37±0.72^bcd
Verteporfin组	403.86±41.09^bcd	6.48±0.68^bcd
F	28.295^＊＊	253.633^＊＊

组别	体质量/g	FBG/（mmol/L）
对照组	432.68±45.09	5.27±0.62
DR组	285.93±29.65^a	23.59±2.61^a
Aca低剂量组	358.76±36.92^b	15.62±1.61^b
Aca中剂量组	382.05±39.02^bc	10.07±1.14^bc
Aca高剂量组	417.25±42.83^bcd	6.37±0.72^bcd
Verteporfin组	403.86±41.09^bcd	6.48±0.68^bcd
F	28.295^＊＊	253.633^＊＊

组别	VEGF/（ng/L）	Ang-2/（μg/L）
对照组	105.32±10.38	6.38±0.66
DR组	163.58±17.14^a	11.92±1.24^a
Aca低剂量组	139.74±13.18^b	9.27±1.01^b
Aca中剂量组	125.07±12.49^bc	8.04±0.91^bc
Aca高剂量组	109.31±11.26^bcd	7.05±0.77^bc
Verteporfin组	110.83±11.75^bcd	7.48±0.78^bcd
F	30.569^＊＊	47.819^＊＊

组别	VEGF/（ng/L）	Ang-2/（μg/L）
对照组	105.32±10.38	6.38±0.66
DR组	163.58±17.14^a	11.92±1.24^a
Aca低剂量组	139.74±13.18^b	9.27±1.01^b
Aca中剂量组	125.07±12.49^bc	8.04±0.91^bc
Aca高剂量组	109.31±11.26^bcd	7.05±0.77^bc
Verteporfin组	110.83±11.75^bcd	7.48±0.78^bcd
F	30.569^＊＊	47.819^＊＊

组别	VEGF	HIF-1α	VCAM-1
对照组	0.42±0.09	0.58±0.06	0.38±0.04
DR组	1.43±0.15^a	1.26±0.12^a	1.36±0.18^a
Aca低剂量组	0.91±0.09^b	0.95±0.09^b	0.99±0.10^b
Aca中剂量组	0.73±0.07^bc	0.73±0.08^bc	0.61±0.07^bc
Aca高剂量组	0.52±0.06^bcd	0.61±0.07^bcd	0.45±0.05^bcd
Verteporfin组	0.59±0.06^bcd	0.63±0.08^bcd	0.43±0.04^bcd
F	95.456^＊＊	57.885^＊＊	103.997^＊＊

金合欢素调节Hippo信号通路对糖尿病视网膜病变大鼠血管生成的影响

Effects of acacetin on angiogenesis in diabetes retinopathy rats by regulating Hippo signaling pathway

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组别	YAP	p-YAP	LATS2	TAZ	TEAD1
对照组	0.53±0.06	0.93±0.09	0.85±0.09	0.73±0.08	0.48±0.06
DR组	1.35±0.14^a	0.35±0.04^a	0.35±0.04^a	1.53±0.16^a	1.29±0.15^a
Aca低剂量组	0.98±0.10^b	0.48±0.05^b	0.98±0.10^b	1.24±0.14^b	0.93±0.09^b
Aca中剂量组	0.75±0.08^bc	0.67±0.07^bc	0.75±0.08^bc	0.95±0.10^bc	0.72±0.07^bc
Aca高剂量组	0.57±0.06^bcd	0.88±0.09^bcd	0.57±0.06^bcd	0.79±0.08^bcd	0.53±0.06^bcd
Verteporfin组	0.55±0.08^bcd	0.87±0.08^bcd	0.55±0.08^bcd	0.81±0.08^bcd	0.58±0.06^bcd
F	76.193^＊＊	65.149^＊＊	52.165^＊＊	47.776^＊＊	73.858^＊＊

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