金丝桃素通过抑制NF-κB信号通路减轻糖尿病肾病足细胞损伤

doi:10.3969/j.issn.1674-8115.2026.01.005

上海交通大学学报（医学版） ›› 2026, Vol. 46 ›› Issue (1): 43-53.doi: 10.3969/j.issn.1674-8115.2026.01.005

金丝桃素通过抑制NF-κB信号通路减轻糖尿病肾病足细胞损伤

韩珍¹, 王浩¹, 苏秀秀²(), 方际¹()

^1.上海中医药大学附属普陀医院肾内科，上海 200062
^2.上海交通大学医学院附属瑞金医院心内科，上海 200025

收稿日期:2025-05-22 接受日期:2025-08-12 出版日期:2026-01-28 发布日期:2026-01-30
通讯作者: 苏秀秀，主治医师，硕士；电子信箱：sxx11901@rjh.com.cn
方　际，副主任医师，硕士；电子信箱：fjzyy2025@163.com。
作者简介:第一联系人：韩珍和苏秀秀负责实验操作、数据采集和分析、论文撰写，方际和王浩参与实验设计与论文的修改。所有作者均阅读并同意了最终稿件的提交。
基金资助:
上海市普陀区卫生健康系统临床特色专科建设项目(2021tszk02)

Hypericin alleviates podocyte injury in diabetic nephropathy by inhibiting the NF-κB signaling pathway

Han Zhen¹, Wang Hao¹, Su Xiuxiu²(), Fang Ji¹()

^1.Department of Nephrology, Putuo Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200062, China
^2.Department of Cardiology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China

Received:2025-05-22 Accepted:2025-08-12 Online:2026-01-28 Published:2026-01-30
Contact: Su Xiuxiu, E-mail: sxx11901@rjh.com.cn
Fang Ji, E-mail: fjzyy2025@163.com.
About author:First author contact:Han Zhen and Su Xiuxiu were responsible for experimental operations, data collection and analysis, and paper writing. Fang Ji and Wang Hao participated in the experimental design and paper revision. All authors have read the final version of the paper and consented to its submission.
Supported by:
Shanghai Putuo District Health System Clinical Specialty Construction Project(2021tszk02)

摘要/Abstract

摘要：

目的·探讨天然化合物金丝桃素（hypericin，HYP）对糖尿病肾病（diabetic nephropathy，DN）足细胞损伤的保护作用及其可能的分子机制。方法·选取18只db/db小鼠构建DN模型，随机分为模型组、db/db+HYP低剂量组（db/db+HYP-L，1.5 mg/kg）、db/db+HYP高剂量组（db/db+HYP-H，3.0 mg/kg），并设6只db/m小鼠为正常对照组。检测各组小鼠空腹血糖（fasting blood glucose，FBG）、胰岛素（insulin，INS）、总胆固醇（total cholesterol，TC）、甘油三酯（triglyceride，TAG）、低密度脂蛋白（low-density lipoprotein，LDL）、高密度脂蛋白（high-density lipoprotein，HDL）、血肌酐（serum creatinine，Scr）、血尿素氮（blood urea nitrogen，BUN）、尿白蛋白/肌酐比值（urinary albumin-to-creatinine ratio，UACR）及炎症因子［肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素-1β（interleukin-1β，IL-1β）、白细胞介素-6（interleukin-6，IL-6）、单核细胞趋化蛋白-1（monocyte chemoattractant protein-1，MCP-1）］水平。经苏木精-伊红（hematoxylin-eosin，HE）、过碘酸-希夫（periodic acid Schiff，PAS）染色以及透射电镜观察各组小鼠肾脏病理变化及足细胞超微结构。免疫组化及蛋白质印迹法（Western blotting）检测各组小鼠肾组织中足细胞标志物Wilms瘤1（Wilms tumor 1，WT-1）的表达。蛋白质印迹法检测肾组织核因子κB（nuclear factor-κB，NF-κB）的表达水平。体外培养人足细胞系，建立高糖诱导足细胞损伤模型，实验分为5组：正常糖组（5.5 mmol/L glucose，NG）、甘露醇组（5.5 mmol/L glucose+24.5 mmol/L mannitol，MA）、高糖组（30 mmol/L glucose，HG）、高糖+HYP 0.05 μg/mL组（HG+0.05 μg/mL HYP）、高糖+HYP 0.20 μg/mL组（HG+0.20 μg/mL HYP），48 h后收集细胞。通过蛋白质印迹法检测WT-1、p-NF-κB p65、NF-κB p65蛋白水平，实时荧光定量PCR（quantitative real-time，qPCR）检测各组人足细胞炎症因子（TNF-α、IL-1β、IL-6、MCP-1）的mRNA水平。结果·与模型组相比，HYP显著降低db/db小鼠FBG、INS、TC、TAG、LDL、Scr、BUN及UACR水平，显著升高HDL水平（均P<0.05），并减轻db/db小鼠足细胞损伤，表现为足突融合现象显著减轻，足突排列重现梳子齿状。体内和体外研究均发现，HYP上调WT-1表达，同时抑制NF-κB p65磷酸化及其介导的炎症因子（TNF-α、IL-1β、IL-6、MCP-1）释放（均P<0.05）。结论·HYP可以减轻db/db小鼠足细胞损伤，其作用可能与上调WT-1表达、抑制NF-κB通路、下调炎症因子表达有关。

关键词: 金丝桃素, 糖尿病肾病, 足细胞, 核因子κB, 炎症反应

Abstract:

Objective ·To investigate the protective effect of the natural compound hypericin (HYP) on podocyte injury in diabetic nephropathy (DN) and its underlying molecular mechanism. Methods ·A total of eighteen db/db mice were selected to establish the DN model and were randomly divided into three groups: the model group, the db/db + HYP low-dose group (db/db+HYP-L, 1.5 mg/kg), and the db/db+HYP high-dose group (db/db+HYP-H, 3 mg/kg). Additionally, six db/m mice were designated as the normal control group. The following parameters were measured in mice from each group: fasting blood glucose (FBG), insulin (INS), total cholesterol (TC), triglyceride (TAG), low-density lipoprotein (LDL), high-density lipoprotein (HDL), serum creatinine (Scr), blood urea nitrogen (BUN), urinary albumin-to-creatinine ratio (UACR), as well as levels of inflammatory cytokines, including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), and monocyte chemoattractant protein-1 (MCP-1). Renal pathological changes and podocyte ultrastructure in mice from each group were examined by hematoxylin-eosin (HE) staining, periodic acid-Schiff (PAS) staining, and transmission electron microscopy. The expression of the podocyte marker Wilms tumor 1 (WT-1) in renal tissues from each group was assessed using immunohistochemistry and Western blotting. The protein expression level of nuclear factor-κB (NF-κB) was also measured by Western blotting. Human podocyte lines were cultured in vitro to establish a high-glucose-induced podocyte injury model. The experiment was divided into five groups: normal glucose (5.5 mmol/L glucose, NG), mannitol (5.5 mmol/L glucose+24.5 mmol/L mannitol, MA), high glucose (30.0 mmol/L glucose, HG), high glucose+0.05 μg/mL HYP, and high glucose+0.20 μg/mL HYP. Cells were harvested after 48 h of incubation. The protein levels of WT-1, phospho-NF-κB p65 (p-NF-κB p65), and NF-κB p65 were determined by Western blotting. The mRNA expression levels of inflammatory cytokines (TNF-α, IL-1β, IL-6, and MCP-1) in human podocytes across all groups were measured by quantitative real-time PCR (qPCR). Results ·Compared with the model group, HYP administration significantly reduced the levels of FBG, INS, TC, TAG, LDL, Scr, BUN, and UACR in db/db mice, while significantly increasing the level of HDL (all P<0.05). Concomitantly, HYP alleviated podocyte injury in db/db mice, as evidenced by a marked reduction in foot process effacement and restoration of a comb-like alignment of the foot processes. Both in vivo and in vitro studies demonstrated that HYP upregulated the expression of WT-1 while concurrently suppressing the phosphorylation of NF-κB p65 and the subsequent release of NF-κB-mediated inflammatory cytokines (TNF-α, IL-1β, IL-6, and MCP-1) (all P<0.05). Conclusion ·HYP alleviates podocyte injury in db/db mice, an effect that may be attributed to the upregulation of WT-1 expression and inhibition of the NF-κB signaling pathway, leading to reduced expression of downstream inflammatory cytokines.

Key words: hypericin (HYP), diabetic nephropathy (DN), podocyte, nuclear factor-kappa B (NF-κB), inflammation

中图分类号:

R692.9

韩珍, 王浩, 苏秀秀, 方际. 金丝桃素通过抑制NF-κB信号通路减轻糖尿病肾病足细胞损伤[J]. 上海交通大学学报（医学版）, 2026, 46(1): 43-53.

Han Zhen, Wang Hao, Su Xiuxiu, Fang Ji. Hypericin alleviates podocyte injury in diabetic nephropathy by inhibiting the NF-κB signaling pathway[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2026, 46(1): 43-53.

图/表 9

表1 引物序列

Tab 1 Primer sequences for qPCR

Gene	Forward primer (5′→3′)	Reverse primer (5′→3′)
TNF-α	GGCAGGTTCTGTCCCTTTCA	CTGTGCTCATGGTGTCTTTTCTG
IL-1β	CTACAGGCTCCGAGATGAACAAC	TCCATTGAGGTGGAGAGCTTTC
IL-6	ACTCACCTCTTCAGAACGAATTG	CCATCTTTGGAAGGTTCAGGTTG
MCP-1	CAGCCAGATGCAATCAATGCC	TGGAATCCTGAACCCACTTCT
β-actin	CATGTACGTTGCTATCCAGGC	CTCCTTAATGTCACGCACGAT

图1 金丝桃素在 db/db 小鼠肾脏和HPCs中的分布Note： Fluorescence intensity in the organs of db/db mice at 4 h after intravenous injection of HYP (5 mg/kg). A. In vivo imaging showed fluorescence intensity. B. Line graph of background-subtracted fluorescence intensity in various organs. ①P=0.021, compared with the heart. C. Fluorescence intensity of HYP in HPCs at different time points (×200). Scale bar=20 μm.

Fig 1 Distribution of HYP in the kidneys of db/db mice and HPCs

图2 db/db 小鼠糖尿病代谢表型的多参数动态分析Note： HYP reduced water intake, urine output, food consumption, and body weight in db/db mice. A. Water intake. B. Urine volume. C. Food intake. D. Body weight. ①P=0.007,②P=0.034,③P=0.023,④P=0.042, compared with the model group (n=6).

Fig 2 Dynamic multi-parameter analysis of diabetic metabolic phenotypes in db/db mice

图3 HYP干预对 db/db 小鼠糖脂代谢的影响Note： HYP reduced insulin levels and ameliorated glucose and lipid metabolic disorders in db/db mice. A. Insulin levels. B. Fasting blood glucose levels. C. Total cholesterol (TC). D. Triglycerides (TAG). E. Low-density lipoprotein (LDL). F. High-density lipoprotein (HDL). ①P<0.001, compared with the db/m group; ②P=0.003,③P=0.027, ④P=0.032,⑤P=0.006,⑥P=0.017,⑦P=0.002, compared with the model group (n=6).

Fig 3 Effect of HYP intervention on glucose and lipid metabolism in db/db mice

图4 HYP对 db/db 小鼠肾功能的改善Note： HYP ameliorated renal function in db/db mice. A. Scr. B. BUN. C. UACR. ①P<0.001, compared with the db/m group; ②P=0.003,③P=0.006,④P=0.026, compared with the model group (n=6).

Fig 4 Hypericin ameliorates renal dysfunction in db/db mice

图5 HE、PAS染色、免疫组化、透射电镜评估肾组织病理学变化Note： HYP ameliorated podocyte injury in db/db mice. A. HE staining showed histopathological changes in glomeruli across groups (×400). B. PAS staining revealed glomerular basement membrane thickness and mesangial matrix expansion in each group (×400). C. WT-1 expression in mouse glomeruli from different groups (×400). D. Transmission electron microscopy showed podocyte foot process effacement (×20 000). Scale bars=20 μm (A, B, C); 500 nm (D).

Fig 5 Renal histopathological changes were evaluated by HE staining, PAS staining, immunohistochemistry, and transmission electron microscopy

图6 HYP对高糖诱导的HPCs细胞活力的影响Note： HYP restored the viability of HPCs downregulated by high-glucose-induced. A. Cell viability of HPCs cultured under normal glucose conditions with different concentrations of HYP. B. Cell viability of the high-glucose-induced HPCs at different time points. C. Cell viability of the high-glucose-induced HPC model after intervention with different concentrations of HYP. ①P=0.037, compared with the 0 group. ②P=0.008, ③P=0.016, ④P=0.009, compared with the NG group.

Fig 6 Effects of HYP on the viability of high-glucose-induced HPCs

图7 HYP下调炎症因子表达Note：Inflammatory cytokine levels across groups. A.TNF-α (pg/mL), IL-1β (pg/mL), IL-6 (pg/mL), and MCP-1(ng/mL). ①P<0.001, compared with the db/m group; ②P<0.001, compared with the model group (n=6). B. The mRNA expression levels of TNF-α, IL-1β, IL-6, and MCP-1. ③P<0.001, compared with the NG group; ④P<0.001, compared with the HG group.

Fig 7 HYP downregulates the expression of inflammatory cytokines

图8 蛋白质印迹法检测HYP对WT-1蛋白及NF-κB信号通路蛋白表达的影响Note： The effect of HYP on the expression of WT-1 protein and NF-κB signaling pathway-related proteins. A. Western blotting analysis of WT-1, p-NF-κB p65, NF-κB p65, and β-actin protein expression. (①P<0.001, compared with the db/m group; ②P=0.003, ③P=0.002, compared with the model group; n=3). B. Western blotting analysis of the protein expression levels of WT-1, p-NF-κB p65, NF-κB p65, and β-actin in HPCs. Quantitative analysis of WT-1, p-NF-κB p65, and NF-κB p65. (④P<0.001, compared with the NG group; ⑤P<0.001, compared with the HG group).

Fig 8 Effect of HYP on WT-1 and NF-κB signaling pathway protein expression analyzed by Western blotting

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金丝桃素通过抑制NF-κB信号通路减轻糖尿病肾病足细胞损伤

Hypericin alleviates podocyte injury in diabetic nephropathy by inhibiting the NF-κB signaling pathway

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