| 马力,童宁,王仲建.丹皮酚通过抑制 TGF-β1/JNK信号通路保护糖尿病视网膜血管功能[J].安徽医药,2025,29(6):1106-1113. |
| 丹皮酚通过抑制 TGF-β1/JNK信号通路保护糖尿病视网膜血管功能 |
| Paeonol ameliorates diabetic retinal vascular function by inhibiting the TGF-β1/JNK signaling pathway |
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| DOI：10.3969/j.issn.1009-6469.2025.06.009 |
| 中文关键词: 丹皮酚 糖尿病视网膜病变 血管功能障碍 TGF-β1/JNK信号通路 血管形成 |
| 英文关键词: Paeonol Diabetic retinopathy Vascular dysfunction |
| 基金项目: |
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| 摘要点击次数: 72 |
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| 中文摘要: |
| 目的研究丹皮酚对糖尿病视网膜血管功能的保护作用及其作用机制。方法 2021年 10月至 2023年 7月， 50只 SD大鼠按随机数字表法分为对照组、糖尿病视网膜病变( DR)组、低剂量丹皮酚组、中剂量丹皮酚组和高剂量丹皮酚组，每组各 10只。 DR组和 3个丹皮酚组大鼠腹腔注射链脲佐菌素构建 DR动物模型。丹皮酚组大鼠经 100、200和 400 mg/kg丹皮酚连续灌胃 12周，每天 1次。对照组和 DR组大鼠灌胃等量生理盐水。视网膜电图，荧光素眼底血管造影术，视网膜消化铺片，伊文思蓝和苏木精 -伊红( HE)染色评价视网膜结构损伤和血管功能障碍。用高糖( 35 mmol/L)处理人视网膜微血管内皮细胞(HRMECs)构建 DR细胞模型。 HRMECs分为五组:正常糖组、高糖组、低剂量丹皮酚组、中剂量丹皮酚组和高剂量丹皮酚组。正常糖组加入 5.5 mmol/L葡萄糖，高糖组和 3个丹皮酚组加入 35 mmol/L葡萄糖。丹皮酚组分别用 10、20和 40 μmol/L丹皮酚处理。以噻唑蓝( MTT)、 5-乙炔基 -2’脱氧尿嘧啶核苷( EdU)、划痕、 Transwell和成管实验分别检测细胞活性、增殖、迁移和血管形成。转化生长因子 β1(TGF-β1)含量及其 mRNA水平用酶联免疫吸附试验和实时荧光定量逆转录聚合酶链反应( qRT-PCR)检测。蛋白质印迹法检测 TGF-β1、c-Jun氨基端激酶( JNK)和磷酸化(p)-JNK蛋白表达水平。结果与对照组相比， DR组大鼠血糖含量显著增加，体质量明显减少； a波振幅、 a波潜时和 b波潜时明显增加，而 b波振幅显著降低；视网膜中血管分支数、无细胞血管数和伊文思蓝含量显著增多；视网膜组织结构松散，神经纤维层伴水肿，内外核层细胞排列不规则，有大量新生血管生成( P<0.001)。与 DR组比较，丹皮酚组糖含量显著降低，体质量明显增加； a波振幅、 a波潜时和 b波潜时明显降低，而 b波振幅显著增加；视网膜中血管分支数、无细胞血管数和伊文思蓝含量显著减少；网膜组织结构较为完整，内外核层细胞排列较整齐，新生血管生成较模型组明显减少( P<0.001)。与正常糖组比较，高糖组 HRMECs细胞活性和 EdU阳性细胞数明显减少，细胞迁移率、迁移细胞数、血管样结构数和血管分支点数明显增多；与高糖组比较，丹皮酚组细胞活性和 EdU阳性细胞数明显增加，细胞迁移率、迁移细胞数、血管样结构数和血管分支点数明显减少( P<0.001)。与对照组［(121.53±10.78)ng/L］和正常糖组［(299.87±15.39)ng/L］比较， DR组大鼠血清［(602.87±10.14)ng/L］及高糖组细胞上清中 TGF-β1含量［(1 002.66±16.53)ng/L］显著增加(均 P<0.001)。与对照组( 1.00±0.03)和正常糖组( 1.00±0.02)比较， DR组大鼠视网膜组织( 3.01±0.23)和高糖组 HRMECs中( 4.52±0.23)TGF-β1的 mRNA表达显著增加，低、中、高剂量丹皮酚处理组视网膜组织中( 2.62±0.19、1.81±0.19、 1.51±0.16)和 HRMECs中( 3.50±0.19、2.03±0.21、1.51±0.12)TGF-β1的 mRNA表达则呈剂量依赖性降低( P<0.001)。与对照组(0.54±0.09、0.14±0.04)和正常糖组( 0.15±0.04、0.29±0.07)比较， DR大鼠视网膜组织( 0.95±0.11、1.07±0.06)和高糖组 HRMECs中( 1.16±0.11、0.98±0.07)TGF-β1、p-JNK蛋白表达显著增加，低、中、高剂量丹皮酚处理组视网膜组织中( 0.45±0.05、0.46±0.06、 0.41±0.08；0.84±0.05、0.48±0.08、0.19±0.06)和 HRMECs中( 0.75±0.11、0.56±0.11、0.42±0.08；0.77±0.04、0.25±0.07、0.27±0.06) TGF-β1、p-JNK蛋白表达则呈剂量依赖性降低( P<0.001)。结论丹皮酚通过抑制 TGF-β1/JNK通路保护糖尿病视网膜血管功能。 |
| 英文摘要: |
| Objective To explore the protective effects of paeonol on diabetic retinal vascular dysfunction and its potential mecha-nism.Methods From October 2021 to July 2023, 50 SD rats were randomly divided into five groups: control, DR model, and low-, me-dium-, and high-dose paeonol groups (10 rats per group). Except for the control group, the other four groups were injected with 60 mg/ kg STZ to establish a DR model. The three paeonol groups were administrated 100, 200 and 400 mg/kg paeonol via intragastric adminis-tration for 12 weeks, once daily. The control and DR model groups received equal volumes of normal saline. Retinal damage and vascu-lar dysfunction were assessed via electroretinography, fluorescein fundus angiography, retinal trypsin digestion, Evans blue staining,and H&E staining. HRMECs were treated with high glucose to establish a DR cell model and divided into five groups: normal glucosegroup, high glucose and three paeonol groups. Cells in the normal glucose group were treated with 5.5 mmol/L glucose, while the otherfour groups were exposed to 35 mmol/L glucose. The three paeonol groups were treated with 10, 20 and 40 μmol/L paeonol. Cell viabili-ty, proliferation, migration, and tube formation were assessed using 3-(4, 5-dimethylthiazole-2) -2, 5-diphenyltetrazolium bromide (MTT), 5-acetylene-2'-deoxyuridine (EdU), wound healing, Transwell, and tube formation assays. Enzyme linked-immunosorbent assay (ELISA) and quantitative reverse-transcription polymerase chain reaction (qRT-PCR) were used to evaluate transforming growth factor -β1 (TGF-β1) expression, and Western blotting was used to determine TGF-β1, c-Jun N-terminal kinase (JNK), and phosphorylated (p)-JNK protein levels.Results The high glucose group exhibited significantly decreased cell viability and fewer EdU-positive cells com-pared to the normal glucose group (P<0.001). Furthermore, the high glucose group showed n increased cell migration rate, a highernumber of migrated cells, enhanced tube-like structure formation, and more vascular branch points (P<0.001). In contrast, the paeonol-treated group demonstrated significantly improved cell viability and a greater number of EdU-positive cells compared to the high glu-cose group (P<0.001). Additionally, the cell migration rate, number of migrated cells, tube-like structure formation, and vascular branch points were significantly reduced in the paeonol-treated group (P<0.001). Serum TGF-β1 levels in the DR group (602.87±10.14 ng/L) and cell culture supernatant TGF-β1 levels in the high glucose group [(1 002.66±16.53) ng/L] were significantly higher than thosein the control group [(121.53±10.78) ng/L] and normal glucose group [(299.87±15.39) ng/L], respectively (P<0.001). Similarly, TGF-β1 mRNA expression in DR group retinal tissues (3.01±0.23) and high glucose-treated HRMECs (4.52±0.23) was markedly elevated com-pared to the control group (1.00±0.03) and normal glucose group (1.00±0.02). Notably, all paeonol treatment groups displayed dose-de-pendent reductions in TGF-β1 mRNA expression in tissues (2.62±0.19, 1.81±0.19, 1.51±0.16) and cells (3.50±0.19, 2.03±0.21, 1.51± 0.12) (P<0.001).TGF-β1 and p-JNK protein expression in DR rat retinal tissues (0.95±0.11, 1.07±0.06) and high glucose-treated HR‐MECs (1.16±0.11, 0.98±0.07) were significantly upregulated compared to controls (0.54±0.09, 0.14±0.04) and the normal glucosegroup (0.15±0.04, 0.29±0.07). However, paeonol treatment induced dose-dependent decreases in TGF-β1 and p-JNK protein levels intissues (0.45±0.05, 0.46±0.06, 0.41±0.08; 0.84±0.05, 0.48±0.08, 0.19±0.06) and cells (0.75±0.11, 0.56±0.11, 0.42±0.08; 0.77±0.04,0.25±0.07, 0.27±0.06) (P<0.001).Conclusion Paeonol improves the diabetic retinal vascular dysfunction by suppressing the TGF-β1/ |
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