熟地黄通过多靶点机制治疗慢性肾脏病

doi:10.12122/j.issn.1673-4254.2026.07.15

摘要/Abstract

摘要：

目的基于网络药理学和分子对接技术，探索熟地黄治疗慢性肾脏病（CKD）的潜在作用机制。方法通过中药系统药理学数据库与分析平台和中药分子机制生物信息学分析工具筛选熟地黄的活性成分及靶点，构建“成分-靶点-通路”网络。采用R软件进行基因本体论和京都基因与基因组百科全书富集分析，揭示熟地黄的作用通路，并利用AutoDock Vina对核心靶点Jun原癌基因（JUN）和信号转导及转录激活因子3（STAT3）与活性成分进行分子对接。动物实验方面，选取8周龄雄性C57BL/6小鼠，随机分为假手术组（Ctrl组）、单侧肾缺血再灌注损伤模型组（UIRI组）及熟地黄干预组（UIRI+RRP组，包括低、中、高剂量组），每组6只。采用夹闭右肾蒂30 min建立UIRI模型，假手术组仅暴露肾脏而不夹闭。干预组于造模前2 d至术后10 d分别给予熟地黄水煎液灌胃（300、600、1200 mg·kg^-1·d^-1），其余各组给予等量生理盐水。术后第30天切除左肾并收集血液及肾组织标本。检测血清肌酐、尿酸、尿素氮、尿蛋白水平及肾组织病理变化（苏木精-伊红染色、马松染色、天狼星红染色），并通过免疫组化分析JUN、STAT3、I型胶原（Collagen I）和III型胶原（Collagen III）表达。结果共筛选25种熟地黄活性成分及307个CKD相关交集靶基因，蛋白质-蛋白质相互作用网络分析确定19个核心靶基因，其中JUN和STAT3为核心节点。分子对接证实熟地黄活性成分与JUN和STAT3高亲和力结合。与Ctrl组相比，UIRI组小鼠血清肌酐、尿酸、尿素氮及尿蛋白水平均升高（P<0.01），肾组织可见肾小管扩张及间质纤维化加重，JUN、STAT3、Collagen I及Collagen III表达上调；与UIRI组相比，UIRI+RRP组上述指标水平均降低（P<0.01），肾组织病理损伤程度减轻，JUN、STAT3、Collagen I及Collagen III表达下调。结论熟地黄通过“多成分-多靶点-多通路”机制，靶向JUN/STAT3信号通路，发挥抗纤维化作用，显著延缓CKD进展，为其潜在临床应用提供了理论依据。

关键词: 熟地黄, 慢性肾脏病, 网络药理学, JUN, STAT3

Abstract:

Objective To explore the mechanisms of Rehmanniae radix preparate (RRP) for alleviating chronic kidney disease (CKD). Methods The active components and targets of RRP were identified using TCMSP and BATMAN-TCM databases. A component-target-pathway network was constructed, followed by GO and KEGG enrichment analyses. Molecular docking was performed between the core targets (JUN and STAT3) and the active compounds using AutoDock Vina. For validation of the results, C57BL/6 mice were randomly divided into sham-operated group, unilateral renal ischemia-reperfusion injury (UIRI) model group, and UIRI+RRP treatment group (n=6). RRP treatment was administered by daily gavage (300, 600 and 1200 mg·kg) from 2 days before to 10 days after surgery. On postoperative day 30, serum creatinine, uric acid, blood urea nitrogen, and urinary protein levels were analyzed, and renal histopathology was assessed using HE, Masson, and Sirius Red staining. Immunohistochemistry was used to detect renal expressions of JUN, STAT3, collagen I, and collagen III. Results A total of 25 active compounds and 307 CKD-related targets were identified, and protein-protein interaction analysis identified 19 hub genes with JUN and STAT3 as the key targets. Molecular docking showed strong binding affinities between the active compounds and JUN/STAT3. The mouse models of UIRI showed significant elevation of serum creatinine, uric acid, blood urea nitrogen, and urinary protein levels with marked tubular injury, interstitial fibrosis, and increased renal expressions of JUN, STAT3, collagen I, and collagen III. RRP treatment significantly reduced these biochemical parameters in the mouse models, improved renal histology, and downregulated JUN, STAT3, and fibrosis-related proteins. Conclusion The ameliorative effect of RRP against renal fibrosis and CKD progression in mice is mediated by a multi-component, multi-target, and multi-pathway mechanism targeting the JUN/STAT3 signaling pathway.

Key words: rehmanniae radix praeparata, chronic kidney disease, network pharmacology, JUN, STAT3

林灶强, 江梦瑶, 冯流畅, 方坤洋, 方萍君, 谭秦湘. 熟地黄通过多靶点机制治疗慢性肾脏病[J]. 南方医科大学学报, 2026, 46(7): 1611-1621.

Zaoqiang LIN, Mengyao JIANG, Liuchang FENG, Kunyang FANG, Pingjun FANG, Qinxiang TAN. Rehmanniae radix preparata alleviates chronic kidney disease in mice through a multi-target mechanism[J]. Journal of Southern Medical University, 2026, 46(7): 1611-1621.

图/表 15

表1 RRP的有效成份

Tab.1 Active components of RRP

Active ingredients	Source	Active ingredients	Source
ID:5742590	TCMSP	ID:45356795	BATMANTCM
ID:3084097	TCMSP	ID:10467	BATMANTCM
ID:5280794	TCMSP	ID:7405	BATMANTCM
ID:5281773	BATMANTCM	ID:6029	BATMANTCM
ID:24981346	BATMANTCM	ID:439685	BATMANTCM
ID:53399246	BATMANTCM	ID:1110	BATMANTCM
ID:54600129	BATMANTCM	ID:10542	BATMANTCM
ID:107848	BATMANTCM	ID:439531	BATMANTCM
ID:11252659	BATMANTCM	ID:11005647	BATMANTCM
ID:5988	BATMANTCM	ID:222284	BATMANTCM
ID:7820	BATMANTCM	ID:6030	BATMANTCM
ID:5280794	BATMANTCM	ID:8215	BATMANTCM
ID:3893	BATMANTCM	ID:13849	BATMANTCM
ID:5460660	BATMANTCM	ID:5320903	BATMANTCM
ID:5280450	BATMANTCM	ID:445638	BATMANTCM
ID:15693864	BATMANTCM	ID:53301851	BATMANTCM
ID:6251	BATMANTCM	ID:445858	BATMANTCM
ID:24721256	BATMANTCM	ID:643801	BATMANTCM
ID:137706142	BATMANTCM	ID:91458	BATMANTCM
ID:499	BATMANTCM	ID:439242	BATMANTCM
ID:15693863	BATMANTCM	ID:3931	BATMANTCM
ID:5705531	BATMANTCM	ID:14486934	BATMANTCM
ID:5281800	BATMANTCM	ID:138107792	BATMANTCM

图1 RRP与CKD重叠靶点的鉴定

Fig.1 Identification of intersecting targets between RRP and CKD. A: Venn diagram of intersecting genes from databases; B: Venn diagram of intersecting target genes between RRP and CKD.

图2 RRP-成份-靶点网络图

Fig.2 RRP-component-target network diagram. Squares represent target genes, octagons represent active components from BATMAN-TCM, hexagons represent active components from TCMSP, and circles represent common active components from both TCMSP and BATMAN-TCM.

图3 PPI网络中核心基因的筛选

Fig.3 Screening of core genes in the PPI network.

表2 拓扑参数评分结果

Tab.2 Topological parameter scoring results

Name	Betweenness	Closeness	Degree	Eigenvector	LAC	Network
JUN	29.251	0.857	30	0.320	14.400	23.626
STAT3	19.715	0.818	28	0.310	14.857	22.161
ESR1	19.287	0.818	28	0.307	14.571	22.076
RELA	13.129	0.783	26	0.293	14.462	20.310
TP53	16.546	0.783	26	0.292	13.538	18.684
NFKB1	9.175	0.720	22	0.253	12.364	15.697
CTNNB1	9.246	0.692	20	0.220	9.600	11.510
FOS	7.160	0.692	20	0.238	11.600	13.429
SRC	8.572	0.692	20	0.223	10.000	11.902
AKT1	5.111	0.667	18	0.211	10.222	11.554
MAPK8	2.760	0.667	18	0.230	12.444	13.466
CREB1	4.860	0.667	18	0.215	10.222	11.166
PRKACA	3.344	0.643	16	0.193	9.000	9.600
AR	6.808	0.643	16	0.172	7.500	8.693
TNF	2.238	0.621	14	0.170	8.571	9.231
IL6	1.352	0.621	14	0.171	9.714	10.462
HSP90AA1	0.556	0.600	12	0.145	8.667	9.455
SIRT1	0.222	0.581	10	0.132	7.200	8.000
NCOA1	0.667	0.563	8	0.099	4.000	4.571

图4 RRP治疗CKD交集靶点基因的GO和 KEGG 富集分析

Fig.4 GO and KEGG enrichment analysis of intersecting target genes for RRP treatment of CKD.

图5 靶点蛋白(JUN)受体与活性成份配体(CID_5280450)的分子对接

Fig.5 Molecular docking of target protein receptors (JUN) with active component ligands (CID_5280450).

图6 靶点蛋白(STAT3)受体与活性成份配体(CID_24721256)的分子对接

Fig.6 Molecular docking of target protein receptors (STAT3) with active component ligands (CID_5280450).

图7 小鼠肌酐、尿素氮、尿酸及尿蛋白水平变化

Fig.7 Changes in in creatinine (A), blood urea nitrogen (B), uric acid (C), and urinary protein (D) levels in mouse models of chronic kidney disease treated with RRP at low, median and high doses (n=6). *P<0.05, **P<0.01, ***P<0.001 vs ctrl group, CKD+RRP (low), CKD+RRP (median), or CKD+RRP (high) (n=6).

图8 小鼠肾组织病理染色

Fig.8 Histopathological staining of mouse kidney tissue (Original magnification: ×400).

图 9 小鼠肾组织病理染色定量分析结果

Fig.9 Quantitative analysis of pathological staining of mouse kidney tissue (n=6). ***P<0.001 vs ctrl group, CKD+RRP (low), CKD+RRP (median), or CKD+RRP (high).

图10 小鼠肾组织JUN和STAT3的IHC染色

Fig.10 Immunohistochemical staining of JUN and STAT3 in mouse kidney tissue (×400).

图11 小鼠肾组织p-JUN和p-STAT3的双荧光染色

Fig.11 Double fluorescence staining of p-JUN and p-STAT3 in mouse kidney tissue (×400).

图12 小鼠肾组织TNF-α和IL-6水平

Fig.12 　Levels of TNF‑α and IL-6 in mousekidney tissue ( (n=6). A: TNF‑α levels in thekidney; B: IL-6 levels in the kidney. *P<0.05vs

图13 小鼠肾组织Collagen I和Collagen III的IHC染色

Fig.13 Collagen I and collagen III staining of mouse kidney tissue (×400).

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