中国临床药理学杂志

楮实子调控PI3K/Akt通路对治疗糖尿病肾病的作用研究

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郭锡春 ^1a , 袁梦梦 ^1a , 刘文山 ^1a , 丁传华 ^1a , 韩秀媛 ^1b

中国临床药理学杂志 | 临床与基础桥接研究 2026,42(11): 1545-1551

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中国临床药理学杂志 |临床与基础桥接研究 2026 , 42 (11) : 1545 -1551

楮实子调控PI3K/Akt通路对治疗糖尿病肾病的作用研究

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郭锡春^1a, 袁梦梦^1a, 刘文山^1a, 丁传华^1a, 韩秀媛^1b

作者信息

^1a.山东第二医科大学附属医院药学部/神经与肿瘤药物研发全国重点实验室，山东潍坊 261041

^1b.山东第二医科大学附属医院感染管理办公室，山东潍坊 261041

通讯作者:

韩秀媛，副主任药师 Tel: （0536）3081222 E-mail: hxy21001@163.com

作者简介:

郭锡春（1988-），男，主管药师，主要从事医院药学和临床药学方面的工作

Research of fructus broussonetiae regulating PI3K/Akt pathway in treatment of diabetic kidney disease

Xi-chun GUO^1a, Meng-meng YUAN^1a, Wen-shan LIU^1a, Chuan-hua DING^1a, Xiu-yuan HAN^1b

Affiliations

^1a.Department of Pharmacy/State Key Laboratory of Neurology and Oncology Drug Development, Affiliated Hospital of Shandong Second Medical University, Weifang 261041, Shandong Province, China

^1b.Department of Infection Management, Affiliated Hospital of Shandong Second Medical University, Weifang 261041, Shandong Province, China

出版时间: 2026-06-17 doi: 10.13699/j.cnki.1001-6821.2026.11.008

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摘要

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目的

探讨楮实子调控磷脂酰肌醇3-激酶/蛋白激酶B（PI3K/Akt）通路对糖尿病肾病（DKD）的作用机制。

方法

用中药系统药理学数据库与分析平台（TCMSP）、人类基因数据库（GeneCards）、在线人类孟德尔遗传数据库（OMIM）及治疗靶点数据库（TTD）筛选活性成分与靶点。通过构建蛋白质互作（PPI）网络及拓扑分析，利用基因本体（GO）和京都基因与基因组百科全书（KEGG）进行富集分析，结合分子对接和分子动力学模拟（MD）验证核心成分与靶点的相互作用。将小鼠MPC5细胞分为对照组（正常培养），模型组（30 mmol·L^-1葡萄糖处理48 h），甘露醇组（在对照组基础上，44.5 mmol·L^-1甘露醇处理48 h），实验组（在模型组基础上，2.5、5.0、15.0、30.0、45.0 μmol·L^-1木犀草素分别处理24 h），阳性药物组（在模型组基础上，100 μmol·L^-1卡托普利处理24 h）。用细胞计数（CCK-8）法检测细胞存活率，用流式细胞术检测细胞凋亡，用酶联免疫吸附测定（ELISA）法测定炎症因子白介素-1β（IL-1β）、IL-6、肿瘤坏死因子（TNF-a）的水平，用蛋白质印迹法检测B细胞淋巴瘤-2基因（Bcl-2）、Bcl-2相关X蛋白（Bax）、半胱氨酸天冬氨酸特异性蛋白酶3（Caspase-3）、热休克蛋白90-α（HSP90AA1）、磷酸化蛋白激酶B（p-Akt）与Akt蛋白表达水平。

结果

共获得楮实子与DKD的交集靶点42个，筛选出蛋白激酶B1（Akt1）、前列腺素内过氧化物合酶2（PTGS2）、肿瘤蛋白53（TP53）、HSP90AA1等13个核心靶点。GO/KEGG富集分析显示，其作用机制主要涉及PI3K/Akt信号通路调控及氧化应激、细胞凋亡等生物过程。分子对接与MD证实，HSP90AA1与木犀草素形成的复合物结构最稳定（结合自由能-34.08 kcal·mol^-1），故选定木犀草素开展后续细胞实验验证。CCK-8显示，当浓度达到30.0 μmol·L^-1时木犀草素作用进入平台期，此浓度用于后续实验。对照组、模型组、甘露醇组、实验组（30.0 μmol·L^-1）和阳性药物组的细胞总凋亡率分别为（6.23±0.40）%、（35.47±3.08）%、（6.42±0.52）%、（11.20±1.46）%和（10.78±1.06）%；对照组、模型组和实验组（30.0 μmol·L^-1）细胞的IL-6分泌水平为（29.52±2.21）、（91.12±6.14）和（36.48±3.65）pg·mL^-1，IL-1β分泌水平为（12.68±1.17）、（47.42±3.49）和（18.08±1.42）pg·mL^-1，TNF-α分泌水平为（23.15±1.81）、（79.35±7.01）和（29.62±2.17）pg·mL^-1；Bcl-2蛋白相对表达水平为1.02±0.11、0.45±0.04和0.92±0.07，Bax蛋白相对表达水平为1.01±0.09、2.18±0.17和1.12±0.11，Caspase-3蛋白相对表达水平为1.03±0.08、2.32±0.17和1.31±0.14；HSP90AA1蛋白相对表达水平为1.00±0.07、1.96±0.12和1.08±0.10，p-Akt/Akt为1.00±0.07、2.25±0.07和1.11±0.06。模型组的上述指标与对照组相比，实验组与模型组相比，在统计学上差异均具有统计学意义（均P<0.001）。

结论

楮实子的活性成分木犀草素可能通过下调HSP90AA1表达，抑制PI3K/Akt信号通路的过度激活，同时减轻炎症反应，从而达到治疗糖尿病肾病的目的。

关键词

楮实子 / 木犀草素 / 糖尿病肾病 / 实验验证

Abstract

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Objective

To explore the mechanisms of action of fructus broussonetiae regulating phosphoinositide 3-kinase/ protein kinase B (PI3K/Akt) in diabetic kidney disease (DKD).

Methods

The traditional Chinese medicine systems pharmacology database and analysis platform (TCMSP), the human gene database (GeneCards), online mendelian inheritance in man (OMIM) and therapeutic target database (TTD) were used to screen the active ingredients and targets. The protein-protein interaction (PPI) network and topology analysis were constructed and enriched using gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG), and molecular docking and molecular dynamics (MD) simulations were used to verify the interactions between the core ingredients and targets. Mouse podocyte cells 5 were divided into control group (without any treatment), model group (treated with 30 mmol·L^-1 glucose for 48 h), mannitol group (treated with 44.5 mmol·L^-1 mannitol for 48 h on the basis of the control group), experimental group (treated with 2.5, 5.0, 15.0, 30.0 and 45.0 μmol·L^-1 luteolin for 24 h, on the basis of the model group) and positive drug group (treated with 100 μmol·L^-1 captopril for 24 h on the basis of the model group). Cell viability were detected by cell counting kit-8 (CCK-8), apoptosis were detected by flow cytometry, levels of inflammatory factors interleukin-1β (IL-1β), IL-6 and tumour necrosis factor (TNF-a) were measured by enzyme-linked immunosorbent assay (ELISA), and B-cell lymphoma-2 gene (Bcl-2), Bcl-2-related X protein (Bax), cysteine aspartate specific proteinase-3 (Caspase-3), heat shock protein 90-α (HSP90AA1), phosphorylated protein kinase B (p-Akt) and Akt protein expression levels were detected by Weatern blot.

Results

A total of 42 intersection targets of fructus broussonetiae and DKD were identified, and 13 core targets were screened out, including protein kinase B1 (Akt1), prostaglandin-endoperoxide synthase 2 (PTGS2), tumour protein 53 (TP53) and HSP90AA1, etc. The GO/KEGG enrichment analysis showed that the mechanism of action was involved in the regulation of PI3K/Akt signalling pathway as well as the oxidative stress, apoptosis, etc. The molecular docking and kinetic simulations confirmed that HSP90AA1 formed the most stable complex with luteolin (binding free energy -34.08 kcal·mol^-1), therefore, luteolin were selected for subsequent cellular experiments. CCK-8 showed that the effect of luteolin entered into a plateau phase when the concentration reached 30.0 μmol·L^-1, and this concentration was used for the subsequent experiments. The total apoptosis rates of the control group, model group, mannitol group, experimental group (30.0 μmol·L^-1) and positive drug group were (6.23±0.40)%, (35.47±3.08)%, (6.42±0.52)%, (11.20±1.46)% and (10.78±1.06)%, respectively; the IL-6 levels of the cells in the control, model and experimental groups (30.0 μmol·L^-1) were (29.52±2.21), (91.12±6.14) and (36.48±3.65) pg·mL^-1, respectively; IL-1β levels were (12.68±1.17), (47.42±3.49) and (18.08±1.42) pg·mL^-1, respectively; and TNF-α levels were (23.15±1.81), (79.35±7.01) and (29.62±2.17) pg·mL^-1, respectively; the relative expression levels of Bcl-2 protein were 1.02±0.11, 0.45±0.04 and 0.92±0.07, respectively; the relative expression levels of Bax protein were 1.01±0.09, 2.18±0.17 and 1.12±0.11, respectively; the relative expression levels of Caspase-3 protein relative expression levels were 1.03±0.08, 2.32±0.17 and 1.31±0.14, respectively. The relative expression levels of HSP90AA1 protein were 1.00±0.07, 1.96±0.12 and 1.08±0.10, respectively; the relative expression levels of p-Akt/Akt were 1.00±0.07, 2.25±0.07 and 1.11±0.06, respectively. The above indexes in the model group were statistically significant when compared with those in the control group, and when compared with those in the experimental and model groups (P<0.001).

Conclusion

The active component luteolin of fructus broussonetiae may reduce the expression of HSP90AA1, inhibit the activation of PI3K/Akt signaling pathway, and reduce the inflammatory response, thus achieving the purpose of treating DKD.

Key words

fructus broussonetiae / luteolin / diabetic kidney disease / experimental verification

引用本文

郭锡春, 袁梦梦, 刘文山, 丁传华, 韩秀媛. 楮实子调控PI3K/Akt通路对治疗糖尿病肾病的作用研究. 中国临床药理学杂志, 2026 , 42 (11) : 1545 -1551 . DOI: 10.13699/j.cnki.1001-6821.2026.11.008

Xi-chun GUO, Meng-meng YUAN, Wen-shan LIU, Chuan-hua DING, Xiu-yuan HAN. Research of fructus broussonetiae regulating PI3K/Akt pathway in treatment of diabetic kidney disease[J]. Chinese Journal of Clinical Pharmacology, 2026 , 42 (11) : 1545 -1551 . DOI: 10.13699/j.cnki.1001-6821.2026.11.008

基金

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神经与肿瘤药物研发全国重点实验室开放课题基金资助项目(SKLSIM-F-2024113)
潍坊市卫生健康委员会科研项目(WFWSJK-2025-056)

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2026年第42卷第11期

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文章信息

doi: 10.13699/j.cnki.1001-6821.2026.11.008

接收时间：2026-04-16
首发时间：2026-08-06
出版时间：2026-06-17

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收稿日期：2026-04-16

基金

神经与肿瘤药物研发全国重点实验室开放课题基金资助项目(SKLSIM-F-2024113)

潍坊市卫生健康委员会科研项目(WFWSJK-2025-056)

作者信息

^1a.山东第二医科大学附属医院药学部/神经与肿瘤药物研发全国重点实验室，山东潍坊 261041

^1b.山东第二医科大学附属医院感染管理办公室，山东潍坊 261041

通讯作者:

韩秀媛，副主任药师 Tel: （0536）3081222 E-mail: hxy21001@163.com

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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