Chinese Journal of Animal Nutrition >

2026 , Vol. 38 >Issue 3: 2203 - 2218

DOI: https://doi.org/10.12418/CJAN2026.177

RESEARCH PAPER

Exploring Potential of Sophora alopecuroides L. to Alleviate Heat Stress in Animals Based on Network Pharmacology and Molecular Docking Technology

  • LIU Qi ,
  • WANG Hairong , *
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  • Key Laboratory of Animal Nutrition and Feed Science in Universities of Inner Mongolia Autonomous Region, College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China
*professor, E-mail:

Received date: 2025-07-23

  Online published: 2026-03-16

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Abstract

This study aimed to explore the potential of Sophora alopecuroides L. to alleviate heat stress in animals using network pharmacology and molecular docking technology. The potential effective active ingredients of Sophora alopecuroides L. were retrieved from the HERB database, and their putative targets were predicted using the SwissTargetPrediction database. A “Sophora alopecuroides L.-potential effective active ingredients-targets” network was constructed and visualized with Cytoscape 3.10.3. Heat stress-related targets were collected from the GeneCards, OMIM and TTD databases. The overlapping targets between the potential effective active ingredient action targets of Sophora alopecuroides L. and heat stress-related targets were identified via a Venn diagram generated on the MicroBioInfo platform. These overlapping targets were uploaded to the STRING database to construct a protein-protein interaction (PPI) network, which was further refined and visualized using Cytoscape 3.10.3. Gene Ontology (GO) functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis for the overlapping targets were performed using the DAVID database. Molecular docking between the top 5 core targets from the PPI network and the potential effective active ingredients of Sophora alopecuroides L. was conducted using PyMOL 3.1.5 and AutoDockTools 1.5.7. The results showed as follows: 1) Sophora alopecuroides L. mainly contains 10 potential effective active ingredients, including cytisine, matrine, sophocarpine, sophoramine, and sophoridine. 2) A total of 276 overlapping targets between the action targets of potential effective active ingredients in Sophora alopecuroides L. and the heat stress-related targets were found, primarily involving protein kinase B1 (AKT1), caspase 3 (CASP3), epidermal growth factor receptor (EGFR), heat shock protein 90 alpha family class A member 1 (HSP90AA1), and matrix metalloproteinase 9 (MMP9). 3) GO functional enrichment analysis yielded 618 entries (P<0.01), comprising 372 biological process (BP) related entries [e.g., positive regulation of mitogen-activated protein kinase (MAPK) cascade, chemical synaptic transmission, and adenylate cyclase-activating adrenergic receptor signaling pathway], 80 cellular component (CC) related entries (e.g., plasma membrane, synapse and cell surface), and 166 molecular function (MF) related entries (e.g., identical protein binding, protein kinase activity, and protein serine/threonine kinase activity). 4) KEGG pathway enrichment analysis revealed 148 entries (P<0.05), mainly including the phosphoinositide 3-kinase (PI3K)- protein kinase B (Akt), cyclic adenosine monophosphate (cAMP), and calcium signaling pathways. 5) Molecular docking confirmed stable binding between the top 5 core targets and the potential effective active ingredients of Sophora alopecuroides L. In conclusion, Sophora alopecuroides L. may alleviate heat stress through potential effective active ingredients such as sophocarpine, sophoramine, sophoridine, and matrine, acting on core targets like AKT1, CASP3, EGFR, HSP90AA1 and MMP9, and by modulating multiple signaling pathways including PI3K-Akt, cAMP, and calcium. This study provides a theoretical foundation for further investigation into the mechanism by which Sophora alopecuroides L. mitigates heat stress.

Key words： Sophora alopecuroides L.; network pharmacology; molecular docking; heat stress; mechanism of action

Cite this article

LIU Qi , WANG Hairong . Exploring Potential of Sophora alopecuroides L. to Alleviate Heat Stress in Animals Based on Network Pharmacology and Molecular Docking Technology[J]. Chinese Journal of Animal Nutrition, 2026 , 38(3) : 2203 -2218 . DOI: 10.12418/CJAN2026.177

近年来,我国畜牧业规模化、集约化的程度不断升高,对养殖环境的要求愈发严格。然而,随着全球变暖现象的日益严峻,热应激(heat stress)现象极为普遍,高温环境中动物产热量超出散热能力,引发体温升高,进而造成生理机能紊乱[1],严重影响动物健康及生产性能,带来巨大经济损失,已成为畜牧生产中亟待解决的关键问题。现有缓解手段多依赖物理降温或单一药物干预[2],但存在成本高、效果局限等问题,亟需开发天然植物活性成分的替代方案。苦豆子(Sophora alopecuroides L.)为多年生草本豆科槐属植物,广泛分布于中国西北部、荒地、固定及半固定沙地和盐碱地,具有良好的抗旱、抗寒和耐风沙能力[3],具有抗菌、抗肿瘤[4]、抗炎[5]以及抗氧化[6]作用。本课题组前期研究结果显示,苦豆子能够在高精料饲粮条件下缓解炎症反应并调节能量代谢[7]。从抗氧化和抗炎的作用机制来看,苦豆子可能具有缓解热应激诱导的细胞损伤的作用。网络药理学基于疾病、基因和药物间复杂的相互关系,构建起多层次网络,从整体视角预测药物靶点,为提高药物研发效率提供创新方法[8]。分子对接技术是目前开发药物的重要方法,通过利用虚拟筛选技术寻找药物作用靶点[9],依据受体自身特征,以及受体与药物分子间的相互作用方式,对二者的结合模式与亲和力进行预测[10]。本研究将通过网络药理学与分子对接技术预测苦豆子的潜在有效活性成分、作用靶点、信号通路以及缓解热应激的作用机制,探究苦豆子缓解热应激的潜质,以期为开发基于苦豆子的天然抗热应激制剂提供数据支持。

1 材料与方法

1.1 数据库

本研究所用到的数据库有本草组鉴(HERB)数据库(http://herb.ac.cn)、SwissTargetPrediction数据库(http://www.swisstargetprediction.ch)、GeneCards数据库(https://www.genecards.org)、OMIM数据库(https://www.omim.org)、TTD数据库(https://ttd.idrblab.cn)、DAVID数据库(https://david.ncifcrf.gov或https://davidbioinformatics.nih.gov)、PubChem数据库(https://pubchem.ncbi.nlm.nih.gov)和PDB数据库(https://www.rcsb.org)。

1.2 苦豆子活性成分的筛选与获取

在HERB数据库以“Sophora alopecuroides”为检索词进行检索,获得苦豆子全部活性成分[11];为了保证口服后药物具备良好的吸收利用率并能顺利通过生物膜屏障,以口服生物利用度(oral bioavailability,OB)≥30%和分子质量(molecular weight,MW)≤500 u作为筛选条件[12],筛选苦豆子潜在有效活性成分,并保存其通用简化分子输入线性输入系统(simplified molecular input line entry system,SMILES)表达式。

1.3 苦豆子潜在有效活性成分相关靶点的获取

在SwissTargetPrediction数据库中,选择“Homo sapiens”作为物种,分别导入苦豆子潜在有效活性成分的SMILES表达式,获取相应的作用靶点。

1.4 热应激相关靶点的筛选与获取

在GeneCards数据库、OMIM数据库、TTD数据库中,以“heat stress”为关键词进行检索[13],通过相关性评分(relevance score)筛选热应激的靶点集。靶点信息采集时间与1.3相同。

1.5 蛋白质-蛋白质相互作用(PPI)网络构建及核心靶点筛选

通过微生信平台(https://www.bioinformatics.com.cn),制作韦恩(Venn)图,得到苦豆子潜在有效活性成分作用靶点与热应激相关靶点的交集靶点。在STRING平台(https://cn.string-db.org)输入获取的交集靶点,设置中等置信度(medium confidence)为0.4,得到PPI网络图,下载其PNG和TSV格式。再利用Cytoscape 3.10.3软件构建更清晰的可视化PPI网络图。

1.6 基因本体(GO)功能富集分析和京都基因与基因组百科全书数据库(KEGG)通路富集分析

利用DAVID数据库对交集靶点分别开展GO功能富集分析和KEGG通路富集分析。其中,GO功能富集分析细分为生物过程(biological process,BP)、细胞组分(cellular component,CC)、分子功能(molecular function,MF),用以注释作用靶蛋白在基因功能层面的具体作用;KEGG通路富集分析聚焦于明确药物治疗疾病所涉及的主要信号通路。完成分析后,按照P值对结果进行排序,选取GO功能富集分析各类别的前10个结果[14],以及KEGG通路富集分析中P<0.05[15]的结果,借助微生信平台进行可视化处理。

1.7 苦豆子-潜在有效活性成分-热应激靶点-信号通路网络图构建

将苦豆子潜在有效活性成分缓解热应激的潜在作用靶点及KEGG通路富集分析得到的前20条关键通路富集基因信息制成Network表格和Tape表格,导入Cytoscape 3.10.3软件绘制苦豆子-潜在有效活性成分-热应激靶点-信号通路网络图。

1.8 分子对接验证

将PPI网络中度值(degree)排名前5的核心靶点作为受体[16],与相应的苦豆子潜在有效活性成分进行分子对接,苦豆子潜在有效活性成分作为配体,其3D结构从PubChem下载,保存其3D格式。核心靶点蛋白的结构从PDB数据库获取,通过设定最新年份和分辨率在1.5~2.0 Å范围内进行筛选,下载蛋白质的PDB格式,利用PyMOL 3.1.5软件将受体结构中配体和水分子删除,得到蛋白质受体。将得到的小分子配体文件格式利用Open Babel 3.1.1处理转化为PDB格式,然后使用AutoDockTools 1.5.7进行分子对接,通过结合能评估配体与受体的结合能力。最后利用PyMoL 3.1.5软件进行可视化处理。

2 结果与分析

2.1 苦豆子的潜在有效活性成分靶点分析

通过HERB数据库检索到苦豆子共有26个活性成分,以MW和OB限定筛选之后,最终得到苦豆子的10个潜在有效活性成分,分别是(-)-9α-羟基槐胺碱、金雀花碱、无刺枣碱A、苦参碱、尼克酸、N-甲基金雀花碱、羟基那可汀、槐果碱、槐胺碱、槐定碱,并获取它们的SMILES表达式(表1)。
表1 苦豆子的潜在有效活性成分

Table 1 Potential effective active ingredients in Sophora alopecuroides L.

分子编号
Molecular No.		 分子名称
Molecular name		 口服生物利用度
OB/%		 分子质量
MW/u		 SMILES表达式
SMILES expression
MOL006570 (-)-9α-羟基槐胺碱
(-)-9alpha-hydroxysophoramine		 35.226 54 262.39 C1CC2CN3C(CC=CC3=O)
C4C2N(C1)CC(C4)O
MOL005945 金雀花碱 Cytisine 69.400 08 190.24 C1C2CNCC1C3=CC=CC
(=O)N3C2
MOL012952 无刺枣碱A Daechualkaloid A 34.937 35 179.22 C1CC2N(C1)C3=
C(CCC3=O)CO2
MOL005944 苦参碱 Matrine 63.774 93 248.36 C1CC2C3CCCN4C3C(CCC4)
CN2C(=O)C1
MOL000421 尼克酸 Nicotinic acid 47.645 29 123.11 C1=CC(=CN=C1)C(=O)O
MOL003632 N-甲基金雀花碱
N-methylcytisine		 76.704 35 204.27 CN1CC2CC(C1)C3=
CC=CC(=O)N3C2
MOL003633 羟基那可汀 Oxynarcotine 56.742 70 431.40 CNCCC1=CC2=C(C(=C1CC(=O)
C3=C(C(=C(C=C3)OC)OC)C
(=O)O)OC)OCO2
MOL003627 槐果碱 Sophocarpine 64.264 33 246.35 C1CC2CN3C(CC=CC3=O)
C4C2N(C1)CCC4
MOL003676 槐胺碱 Sophoramine 42.161 88 244.33 C1CC2CN3C(=O)C=CC=
C3C4C2N(C1)CCC4
MOL003680 槐定碱Sophoridine 60.070 33 248.36 C1CC2C3CCCN4C3C(CCC4)
CN2C(=O)C1
在SwissTargetPrediction数据库,利用10个潜在有效活性成分的SMILES表达式,检索各成分相应的靶点,最后汇总整合得到995个靶点,经过去重得到420个作用靶点。

2.2 苦豆子-潜在有效活性成分-作用靶点网络图构建

将上面得到的10个潜在有效活性成分及相应作用靶点导入Cytoscape 3.10.3软件,构建苦豆子-潜在有效活性成分-作用靶点网络图(图1)。经Cytoscape 3.10.3中的“Analyze Network”工具分析可知,该网络图中包括431个节点(1种药物、10种潜在有效活性成分和420个作用靶点)和1 005条边。
图1 苦豆子-潜在有效活性成分-作用靶点网络图

仅注释度值排名前50的节点 Only the top 50 nodes ranked by annotation degree values。Sophoramine:槐胺碱;Sophocarpine:槐果碱;Cytisine:金雀花碱;(-)-9alpha-hydroxysophoramine:(-)-9α-羟基槐胺碱;Sophoridine:槐定碱;Oxynarcotine:羟基那可汀Oxynarcotine: Hydroxynarcotine;N-methylcytisine:N-甲基金雀花碱;Matrine:苦参碱;Daechualkaloid A:无刺枣碱A;Nicotinic acid:尼克酸;SRD5A2:5α-还原酶 2 steroid 5-alpha-reductase 2;PARP1:多聚ADP核糖聚合酶1 poly ADP-ribose polymerase 1;CHRNA4:烟碱型乙酰胆碱受体α4亚基 cholinergic receptor nicotinic alpha 4 subunit;CHRNB2:烟碱型乙酰胆碱受体β2亚基 cholinergic receptor nicotinic beta 2 subunit;ROCK2:Rho相关蛋白激酶2 Rho-associated coiled-coil containing protein kinase 2;HTR3A:5-羟色胺3A受体 5-hydroxytryptamine receptor 3A;SLC6A3:多巴胺转运体 solute carrier family 6 member 3;CHRM2:毒蕈碱型乙酰胆碱受体 M2 cholinergic receptor muscarinic 2;CYP19A1:芳香化酶 cytochrome P450 family 19 subfamily A member 1;BRD4:溴结构域蛋白4 bromodomain containing 4;JAK2:Janus激酶2 Janus kinase 2;ESR2:雌激素受体β estrogen receptor 2;HTR2B:5-羟色胺2B受体 5-hydroxytryptamine receptor 2B;DPP9:二肽基肽酶9 dipeptidyl peptidase 9;HTR1F:5-羟色胺1F受体 5-hydroxytryptamine receptor 1F;ROCK1:Rho相关蛋白激酶1 Rho-associated coiled-coil containing protein kinase 1;DRD1:多巴胺D1受体 dopamine receptor D1;CHRM5:毒蕈碱型乙酰胆碱受体M5 cholinergic receptor muscarinic 5;CHRM4:毒蕈碱型乙酰胆碱受体M4 cholinergic receptor muscarinic 4;CHRM3:毒蕈碱型乙酰胆碱受体M3 cholinergic receptor muscarinic 3;PDE7A:磷酸二酯酶7A phosphodiesterase 7A;MALT1:黏膜相关淋巴组织淋巴瘤转运蛋白1 mucosa associated lymphoid tissue lymphoma translocation protein 1;PRMT3:蛋白质精氨酸甲基转移酶3 protein arginine methyltransferase 3;TYK2:酪氨酸激酶2 tyrosine kinase 2;JAK3:Janus激酶3 Janus kinase 3;PBK:PDZ结合激酶 PDZ binding kinase;PIM2:丝氨酸/苏氨酸激酶PIM2 pim-2 proto-oncogene, serine/threonine kinase;PARP2:多聚ADP核糖聚合酶2 poly ADP-ribose polymerase 2;CHRNA7:烟碱型乙酰胆碱受体α7亚基 cholinergic receptor nicotinic alpha 7 subunit;SRD5A1:5α-还原酶1 steroid 5-alpha-reductase 1;DRD5:多巴胺D5受体 dopamine receptor D5;TACR1:速激肽1受体 tachykinin receptor 1;DPP8:二肽基肽酶 8 dipeptidyl peptidase 8;JAK1:Janus激酶1 Janus kinase 1;ADRA2B:α2B肾上腺素能受体 adrenoceptor alpha 2B;DRD2:多巴胺D2受体 dopamine receptor D2;SLC6A4:5-羟色胺转运体 solute carrier family 6 member 4;CHRNA3:烟碱型乙酰胆碱受体α3亚基 cholinergic receptor nicotinic alpha 3 subunit;CHRNB4:烟碱型乙酰胆碱受体β4亚基 cholinergic receptor nicotinic beta 4 subunit;HSP90AA1:热休克蛋白90αA1 heat shock protein 90 alpha family class A member 1。

Fig.1 Network diagram of Sophora alopecuroides L.-potential effective active ingredients-targets of action

2.3 热应激相关靶点分析

在GeneCards、OMIM、TTD数据库中,以“heat stress”为关键词进行检索,分别得到10 621、143、4个相关靶点,再将从GeneCards数据库得到的靶点通过筛选相关性评分≥1.767 356 873(中位数),得到5 310个相关靶点。将从这3个数据库得到的靶点合并去重,最终得到5 401个热应激相关靶点。

2.4 苦豆子潜在有效活性成分作用靶点与热应激相关靶点的交集靶点分析

由图2可知,苦豆子潜在有效活性成分作用靶点与热应激相关靶点一共存在276个交集靶点。这些多效性生物学靶点的协同作用为缓解热应激症状提供了重要的分子干预基础,显示出苦豆子通过多靶点调控热应激的潜在价值。
图2 苦豆子潜在有效活性成分作用靶点与热应激相关靶点的韦恩图

Fig.2 Venn diagram of between action targets of potential effective active ingredients in Sophora alopecuroides L. and heat stress-related targets

2.5 苦豆子治疗热应激潜在作用靶点的PPI网络构建

通过STRING平台,对276个交集靶点构建PPI网络,设置中等置信度为0.4,得到初步PPI网络图(图3),并下载TSV格式。将TSV文件导入Cytoscape 3.10.3,利用“Analyze Network”工具分析计算出各节点的度值,再按照度值对关键靶点进行排序,得到可视化PPI网络图(图4)。最后利用Cytoscape 3.10.3中的CytoNCA分析出度值、介度中心性(betweenness)、接近中心性(closeness),筛选出排名靠前的10个核心靶点,分别是蛋白激酶B1(AKT1)、半胱氨酸天冬氨酸蛋白酶3(CASP3)、表皮生长因子受体(EGFR)、热休克蛋白90αA1(HSP90AA1)和基质金属蛋白酶9(MMP9)、前列腺素内过氧化物合酶2(PTGS2)、糖原合成酶激酶3β(GSK3B)、淀粉样前体蛋白(APP)、Toll样受体4(TLR4)及人表皮生长因子受体2(ERBB2)。
图3 初步PPI网络图

Fig.3 Preliminary PPI network diagram

图4 可视化PPI网络图

网络中节点越大、颜色越深代表度值越大,该节点在该网络中越重要 The larger the node in the network, the darker the color, and the larger the degree value, the more important the node is in the network。仅注释图中关键节点 Annotate only the key nodes in the diagram。AKT1:蛋白激酶B1 protein kinase B1;CASP3:半胱氨酸天冬氨酸蛋白酶3 caspase 3;EGFR:表皮生长因子受体 epidermal growth factor receptor;HSP90AA1:热休克蛋白90αA1 heat shock protein 90 alpha family class A member 1;MMP9:基质金属蛋白酶9 matrix metalloproteinase 9;PTGS2:前列腺素内过氧化物合酶2 prostaglandin-endoperoxide synthase 2;GSK3B:糖原合成酶激酶3β glycogen synthase kinase 3 beta;APP:淀粉样前体蛋白 amyloid precursor protein;TLR4:Toll样受体4 Toll-like receptor 4;ERBB2:人表皮生长因子受体2 Erb-b2 receptor tyrosine kinase 2;MDM2:鼠双微体2 蛋白 mouse double minute 2 homolog;PARP1:多聚ADP核糖聚合酶1 poly (ADP-ribose) polymerase 1;ACE:血管紧张素转换酶 angiotensin-converting enzyme;CDK2:细胞周期蛋白依赖性激酶2 cyclin-dependent kinase 2;MMP2:基质金属蛋白酶2 matrix metalloproteinase 2;KDR:血管内皮生长因子受体2 kinase insert domain receptor;CDC42:细胞分裂周期蛋白42 cell division cycle 42;HDAC1:组蛋白去乙酰化酶1 histone deacetylase 1;JAK2:Janus激酶2 Janus kinase 2;MAOA:单胺氧化酶A monoamine oxidase A;PTPRC:蛋白酪氨酸磷酸酶受体C protein tyrosine phosphatase receptor type C;MAOB:单胺氧化酶B monoamine oxidase B;CDK1:细胞周期蛋白依赖性激酶1 cyclin-dependent kinase 1;PIK3CA:磷脂酰肌醇3-激酶催化亚基α phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha;CTSB:组织蛋白酶B cathepsin B;CCNA2:细胞周期蛋白A2 cyclin A2;NR3C1:核受体亚家族3C成员1 nuclear receptor subfamily 3 group C member 1;MCL1:髓细胞白血病1蛋白 myeloid cell leukemia 1;MAPK14:丝裂原活化蛋白激酶14 mitogen-activated protein kinase 14;ITGB1:整合素β1 integrin beta 1;DRD2:多巴胺D2受体 dopamine receptor D2;CHEK1:细胞周期检测点激酶1 checkpoint kinase 1;ABL1:ABL原癌基因1 ABL proto-oncogene 1;AR:雄激素受体 androgen receptor。

Fig.4 Visualized PPI network diagram

2.6 GO功能富集分析与KEGG通路富集分析

GO功能富集分析共得到618个条目(P<0.01),其中BP有372个相关条目,主要包括丝裂原活化蛋白激酶(MAPK)级联的正向调控、对异生物质刺激的反应、蛋白质磷酸化、化学突触传递、腺苷酸环化酶激活的肾上腺素能受体信号通路等;CC有80个相关条目,主要包括质膜、突触、细胞表面、树突、突触后膜等;MF有166个相关条目,主要包括相同蛋白质结合、蛋白激酶活性、蛋白丝氨酸/苏氨酸激酶活性、组蛋白H3Y41激酶活性以及蛋白丝氨酸激酶活性等。对GO富集分析结果按P值排序后,选取前10位数据,导入微生信平台生成BP、CC、MF三合一双侧条形图(图5)。
图5 GO功能富集分析图

Biological process:生物过程;Cellular component:细胞组分;Molecular function:分子功能;positive regulation of MAPK cascade:丝裂原活化蛋白激酶级联的正向调控;response to xenobiotic stimulus:对异生物质刺激的反应;protein phosphorylation:蛋白质磷酸化;chemical synaptic transmission:化学突触传递;adenylate cyclase-activating adrenergic receptor signaling pathway:腺苷酸环化酶激活的肾上腺素能受体信号通路;G protein-coupled receptor signaling pathway;coupled to cyclic nucleotide second messenger:与环核苷酸第2信使偶联的G蛋白偶联受体信号通路;proteolysis:蛋白质水解;cellular response to amyloid-beta:细胞对淀粉样β的应答;peptidyl-serine phosphorylation:肽基丝氨酸磷酸化;intracellular signal transduction:细胞内信号转导;plasma membrane:质膜;synapse:突触;cell surface:细胞表面;dendrite:树突;postsynaptic membrane:突触后膜;membrane raft:膜筏;presynaptic membrane:突触前膜;cytoplasm:细胞质;neuronal cell body:神经元胞体;protein-containing complex:含蛋白质的复合物;identical protein binding:相同蛋白质结合;protein kinase activity:蛋白激酶活性;protein serine/threonine kinase activity:蛋白质丝氨酸/苏氨酸激酶活性;histone H3Y41 kinase activity:组蛋白H3Y41激酶活性;histone H2AXY142 kinase activity:组蛋白H2AXY142激酶活性;enzyme binding:酶结合;protein serine kinase activity:蛋白质丝氨酸激酶活性;protein tyrosine kinase activity:蛋白质酪氨酸激酶活性;ATP binding:ATP结合;endopeptidase activity:内肽酶活性。

Fig.5 GO functional enrichment analysis chart

KEGG通路富集分析共得到148个条目(P<0.05),根据P值大小,将前20个关键条目通过微生信平台制成气泡图,主要包括神经活性配体-受体相互作用、钙信号通路、癌症中的蛋白聚糖、血清素能突触、多巴胺能突触、磷脂酰肌醇3激酶-蛋白激酶B(PI3K-Akt)信号通路以及环磷酸腺苷(cAMP)信号通路(cAMP signaling pathway)等。这些通路可能是苦豆子治疗热应激的潜在通路(图6)。
图6 KEGG通路富集分析图

Neuroactive ligand-receptor interaction:神经活性配体-受体相互作用;Calcium signaling pathway:钙信号通路;Proteoglycans in cancer:癌症中的蛋白聚糖;Serotonergic synapse:血清素能突触;Pathways in cancer:癌症中的通路;Focal adhesion:黏着斑;Prostate cancer:前列腺癌;PI3K-Akt signaling pathway:磷脂酰肌醇3激酶-蛋白激酶B信号通路;Dopaminergic synapse:多巴胺能突触;Apoptosis:细胞凋亡;Lipid and atherosclerosis:脂质与动脉粥样硬化;Endocrine resistance:内分泌抵抗;Fluid shear stress and atherosclerosis:流体剪切应力与动脉粥样硬化;cAMP signaling pathway:环磷酸腺苷信号通路;cGMP-PKG signaling pathway:环鸟苷酸依赖性蛋白激酶信号通路;Insulin resistance:胰岛素抵抗;VEGF signaling pathway:血管内皮生长因子信号通路;AGE-RAGE signaling pathway in diabetic complications:糖尿病并发症中的AGE-RAGE信号通路;Chemical carcinogenesis-receptor activation:化学致癌作用-受体激活;Neurotrophin signaling pathway:神经营养因子信号通路。图7同 the same as Fig.7。

Fig.6 KEGG pathway enrichment analysis chart

2.7 苦豆子-潜在有效活性成分-热应激靶点-信号通路网络图构建

通过Cytoscape 3.10.3软件绘制出苦豆子-潜在有效活性成分-热应激靶点-信号通路网络图(图7),并利用“Network Analyze”工具分析。结果显示,该网络图中包括308个节点和999条边。
图7 苦豆子-潜在有效活性成分-热应激靶点-信号通路网络图

Fig.7 Network diagram of Sophora alopecuroides L.-potential effective active ingredients-heat stress targets-signaling pathways

2.8 分子对接验证结果

选取度值居于前5的核心靶点AKT1、CASP3、EGFR、HSP90AA1、MMP9,根据图1的苦豆子-潜在有效活性成分-作用靶点网络图,分别与相应的苦豆子潜在有效活性成分进行分子对接,结果如表2所示。通常情况下,当配体与受体之间的结合能低于-20.92 kJ/mol时,可判定该配体与受体展现出较为良好的结合能力[17];而当结合能进一步降低至-29.29 kJ/mol以下时,则表明二者之间形成了具有显著强度的结合作用[18-19]。由表2可知,5个核心靶点与相应的潜在有效活性成分均具有良好的结合能力,根据结合能的大小,槐果碱与HSP90AA1的结合能力最强,槐定碱、苦参碱、槐胺碱与HSP90AA1结合能力较槐果碱次之,但均具有显著强度的结合能力。(-)-9α-羟基槐胺碱与AKT1,金雀花碱与AKT1、EGFR,无刺枣碱A与EGFR,尼克酸与HSP90AA1,N-甲基金雀花碱与CASP3、EGFR,羟基那可汀与EGFR、MMP9均具有良好的结合能力。将这13个对接结果全部进行可视化处理,可视化结果见图8。通过可视化结果可以发现,除了槐果碱-HSP90AA1的对接结果以外全部具有1条及以上的氢键,槐果碱-HSP90AA1对接的结合能最高,具有1个盐桥和3个疏水作用力。
表2 苦豆子潜在有效活性成分与核心靶点的结合能力

Table 2 Binding ability of active ingredients in Sophora alopecuroides L. and core targets

序号
Serial number		 配体
Ligand		 受体
Receptor		 PDB数据库编号
PDB database ID		 结合能
Binding energy/
(kJ/mol)
A (-)-9α-羟基槐胺碱
(-)-9alpha-hydroxysophoramine		 蛋白激酶B1 AKT1 5AAR -28.70
B
金雀花碱 Cytisine		 蛋白激酶B1 AKT1 5AAR -27.41
C 表皮生长因子受体 EGFR 6TFV -24.39
D 无刺枣碱A Daechualkaloid A 表皮生长因子受体 EGFR 6TFV -26.86
E 苦参碱 Matrine 热休克蛋白90αA1 HSP90AA1 7S8Y -35.06
F 尼克酸 Nicotinic acid 热休克蛋白90αA1 HSP90AA1 7S8Y -25.06
G
N-甲基金雀花碱
N-methylcytisine		 半胱氨酸天冬氨酸蛋白酶3 CASP3 1RHJ -24.52
H 表皮生长因子受体 EGFR 6TFV -24.31
I
羟基那可汀Oxynarcotine		 表皮生长因子受体 EGFR 6TFV -28.45
J 基质金属蛋白酶9 MMP9 5UE4 -21.55
K 槐果碱 Sophocarpine 热休克蛋白90αA1 HSP90AA1 7S8Y -35.56
L 槐胺碱 Sophoramine 热休克蛋白90αA1 HSP90AA1 7S8Y -33.64
M 槐定碱Sophoridine 热休克蛋白90αA1 HSP90AA1 7S8Y -35.15
图8 苦豆子潜在有效活性成分与核心靶点的分子对接结果

Fig.8 Molecular docking results of active ingredients in Sophora alopecuroides L. and core targets

3 讨论

3.1 苦豆子缓解动物热应激的潜在作用基础

以OB≥30%和MW≤500 u作为筛选条件,得到(-)-9α-羟基槐胺碱、金雀花碱、无刺枣碱A、苦参碱、尼克酸、N-甲基金雀花碱、羟基那可丁、槐果碱、槐胺碱以及槐定碱这10种潜在有效活性成分进行后续分析。根据研究报道,金雀花碱具有抗炎[20]、抗肿瘤[21]等作用,苦参碱具有抗炎[22]、抗病毒[23]、保护神经以及抗多种肿瘤的作用[24]。尼克酸,又名烟酸,具有抗炎、抗氧化、调节脂质代谢等作用[25],有研究表明,当奶牛发生热应激时补充尼克酸会使奶牛皮肤毛细血管舒张增加,促进散热,同时降低直肠温度,提高奶牛乳腺上皮细胞热休克蛋白27(HSP27)、热休克蛋白70(HSP70)的表达,从而缓解热应激[26]。槐果碱具有抗肿瘤[27]、抑制炎症与凋亡、抗氧化应激[28]等作用。槐胺碱具有镇痛、抗炎作用[29]等。槐定碱是一种天然的抗凋亡、抗炎与抗氧化剂[30]。这些潜在有效活性成分均有抗炎、抗氧化及调节代谢等特性,为缓解热应激提供了潜在作用基础。

3.2 苦豆子缓解热应激的核心靶点及作用机制

分子对接结果显示,苦豆子的10种潜在有效活性成分与核心靶点AKT1、CASP3、EGFR、HSP90AA1、MMP9对接的结合能均小于-20.92 kJ/mol,且可视化结果显示均具有稳定的分子间作用力。结合2个预测结果表明,苦豆子可能通过作用于AKT1、CASP3、EGFR、HSP90AA1、MMP9等靶点缓解热应激。AKT1可以通过对转录因子进行磷酸化修饰,调控促凋亡基因或抗凋亡基因的表达,在细胞存活过程中发挥重要的作用[31]。有研究表明,AKT1是热应激下热休克蛋白1(HSP1)激活的重要因子[32]。CASP3是细胞凋亡通路中的关键分子靶点,其激活标志着细胞进入不可逆转的凋亡阶段[33]。本研究表明,苦豆子可能通过靶向CASP3缓解热应激诱导的细胞凋亡。EGFR在皮肤屏障角质形成细胞的增殖、分化、迁移及角化进程中发挥关键调控作用,是维系皮肤稳态与表皮发育调控的重要因子[34]。有研究表明,当热应激发生时,猪小肠受到损伤程度与EGFR表达下调相关[35]。哺乳动物依赖热休克蛋白(HSPs)来调节体温和应对热应激[36],在热应激条件下,HSPs通过介导蛋白质的折叠、重折叠过程,调控蛋白质聚集并拮抗潜在的破坏性相互作用[37]。
研究证实,热休克蛋白90(HSP90)基因亚家族成员在维持细胞蛋白质稳态、抑制细胞凋亡、调控转录进程等方面均扮演关键角色[38]。HSP90AA1为HSP90α家族成员1,在启动热应激反应中起着关键作用[39],因此苦豆子中潜在有效活性成分苦参碱、尼克酸、槐定碱、槐果碱、槐胺碱均有可能通过靶向HSP90AA1来缓解动物热应激。MMP9的活性在诸多生物学进程中占据核心地位,涵盖骨骼重建、骨量动态调控以及肿瘤细胞的侵袭迁移等[40]。有研究表明,抑制MMP9的表达能够有效减少中性粒细胞和嗜酸性粒细胞在气道内的浸润,进而减轻局部炎症反应[41],这提示苦豆子中的潜在有效活性成分羟基那可汀可能通过靶向MMP9来缓解热应激给动物带来的机体损伤。

3.3 GO功能富集与KEGG通路富集的生物学意义

根据GO功能富集分析和KEGG通路富集分析结果,核心靶点显著富集在神经活性配体-受体相互作用、钙信号通路、PI3K-Akt信号通路以及cAMP信号通路等多条信号通路上。
在P值排名前20的通路中,PI3K-Akt信号通路是细胞内重要的信号转导通路,在调节细胞迁移、增殖、分化和凋亡中起重要作用。有研究表明,PI3K-Akt信号通路能阻止不同凋亡刺激信号诱导的细胞凋亡,并且在多种细胞凋亡中促进细胞存活[42]。He等[43]研究表明,当动物发生热应激时,PI3K-Akt信号通路可以介导核因子E2相关因子2(Nrf2)/血红素加氧酶-1(HO-1)抗氧化信号通路,缓解热应激诱导的肠上皮屏障功能障碍。
钙作为钙信号通路(calcium signaling pathway)关键信号元件,参与调控细胞分化、增殖与运动、细胞凋亡、分泌、兴奋、收缩及神经元可塑性等多种基本生理过程,钙离子(Ca2+)作为细胞内多种钙信号通路转导的核心调控因子,其可对下游基因表达进行调控,并参与多种细胞功能的调节过程[44]。有研究表明,热应激可促使卵巢颗粒细胞细胞膜上的钙离子进入通道及钙离子释放通道开放,导致细胞内钙离子浓度升高,进而引起钙/钙调蛋白依赖性蛋白激酶2(CAMK2)的蛋白表达上调,激活下游信号通路,诱导细胞凋亡,影响卵巢颗粒细胞功能,最终调控绵羊的繁殖性能[45]。
cAMP信号通路是第2信使系统,参与多种复杂重要的生理过程[46]。cAMP是体内关键的第2信使分子,蛋白激酶A(PKA)作为cAMP的核心效应分子,激活后可直接作用于靶蛋白环磷酸腺苷反应元件结合蛋白(CREB),CREB磷酸化后,进一步调控下游基因的转录过程[47]。郑秦文等[48]研究表明,不同热应激强度下,小鼠乳腺上皮细胞cAMP水平上升,提示热应激激活胞外信号分子,促使ATP环化,表明该信号通路被激活。

3.4 分子对接验证结果分析

分子对接结果表明,苦豆子潜在有效活性成分与排名前5的核心靶点具有较好的结合能力,从原子层面验证了网络药理学的可靠性和准确性,证明了苦豆子潜在有效活性成分在相应作用靶点上具有显著作用。

4 结论

本研究结果表明,苦豆子可能通过槐果碱、槐胺碱、槐定碱、苦参碱等潜在有效活性成分作用于AKT1、CASP3、EGFR、HSP90AA1、MMP9等核心靶点,调控PI3K-Akt、cAMP、钙等多条信号通路缓解动物热应激,该结果可为进一步探究苦豆子缓解热应激的机制提供理论依据。

References

Publishing order | Descend order by publishing year | Descend order by cited within
[1]
郭伟. 夏季奶牛热应激的危害及其防治[J]. 中国畜牧业, 2025(2):102-103.

GUO W. Hazards and prevention of heat stress in dairy cows in summer[J]. China Animal Industry, 2025(2):102-103. (in Chinese)

[2]
黎益图, 邓菊芬, 覃芳娇, 等. 牛热应激的危害及防控措施[J]. 山东畜牧兽医, 2025, 46(5):64-66.

LI Y T, DENG J F, QIN F J, et al. Hazards and prevention measures of heat stress in cattle[J]. Shandong Journal of Animal Science and Veterinary Medicine, 2025, 46(5):64-66. (in Chinese)

[3]
冯平, 麻文静, 沈娜, 等. 苦豆子药理及营养作用的研究进展[J]. 榆林学院学报, 2024, 34(2):19-23.

FENG P, MA W J, SHEN N, et al. Progress of research on the pharmacological and nutritional effects of Sophora alopecuroide[J]. Journal of Yulin University, 2024, 34(2):19-23. (in Chinese)

[4]
梁磊, 王晓燕, 张绪慧, 等. 苦豆子生物碱抗结肠腺癌细胞株SW620的作用筛选[J]. 中药材, 2008, 31(6):866-869.

LIANG L, WANG X Y, ZHANG X H, et al. Screening on alkaloid of Sophora alopecuraides against adenocarcinoma of colon cell line SW620 in vitro[J]. Journal of Chinese Medicinal Materials, 2008, 31(6):866-869. (in Chinese)

[5]
王琳琳, 周娅, 梁锦屏, 等. 槐定碱预防小鼠内毒素性肝损伤的作用及对肝脏CD14、TLR4表达的影响[J]. 宁夏医科大学学报, 2009, 31(6):709-712,722.

WANG L L, ZHOU Y, LIANG J P, et al. Effect of sophoridine on acute liver injury in mice with endotoxemia and its impact on expression of CD14 and TLR4[J]. Journal of Ningxia Medical University, 2009, 31(6):709-712,722. (in Chinese)

[6]
ZHU Y C, WANG Q Y, GUO W Y, et al. Screening and identification of salt-tolerance genes in Sophora alopecuroides and functional verification of SaAQP[J]. Planta, 2021, 254(4):77.

DOI

[7]
安雅雯. 苦豆子对高精料日粮引起绵羊肝脏炎性损伤的调控机制[D].博士学位论文. 呼和浩特: 内蒙古农业大学, 2024.

AN Y W. Regulatory mechanism of Sophora alopecuroides on inflammatory injury of sheep liver caused by a high-concentrate diet[D].Ph.D.Thesis. Hohhot: Inner Mongolia Agricultural University, 2024. (in Chinese)

[8]
李泮霖, 苏薇薇. 网络药理学在中药研究中的最新应用进展[J]. 中草药, 2016, 47(16):2938-2942.

LI P L, SU W W. Recent progress in applying network pharmacology to research of Chinese materia medica[J]. Chinese Traditional and Herbal Drugs, 2016, 47(16):2938-2942. (in Chinese)

[9]
刘玉甜, 赵诗雨, 吕邵娃. 基于分子对接的计算机虚拟筛选技术在新药发现中的应用进展[J]. 化学工程师, 2020, 34(2):59-63.

LIU Y T, ZHAO S Y, LV S W. Application of virtual screening technology based on molecular docking in drug discovery[J]. Chemical Engineer, 2020, 34(2):59-63. (in Chinese)

[10]
杨欣, 李亚辉, 钱海兵, 等. 基于网络药理学及分子对接分析熊果酸抗类风湿性关节炎的分子机制[J]. 中国实验方剂学杂志, 2018, 24(18):207-214.

YANG X, LI Y H, QIAN H B, et al. Molecular mechanism of ursolic acid against rheumatoid arthritis based on network pharmacology and molecular docking[J]. Chinese Journal of Experimental Traditional Medical Formulae, 2018, 24(18):207-214. (in Chinese)

[11]
FANG S S, DONG L, LIU L, et al. HERB:a high-throughput experiment- and reference-guided database of traditional Chinese medicine[J]. Nucleic Acids Research, 2021, 49(D1):D1197-D1206.

[12]
范秋雨, 武建文, 李春晓, 等. 苦豆子抗炎机制的网络药理学分析及试验验证[J]. 中国畜牧兽医, 2023, 50(7):2951-2965.

DOI

FAN Q Y, WU J W, LI C X, et al. Network pharmacology analysis of anti-inflammatory mechanism of Sophora alopecuroides and experimental validation[J]. China Animal Husbandry & Veterinary Medicine, 2023, 50(7):2951-2965. (in Chinese)

[13]
刘逸雷, 丁康宁, 邱文才, 等. 基于网络药理学探讨柴胡口服液调治家禽热应激的分子机理[J]. 黑龙江畜牧兽医, 2021(16):109-115,153-154.

LIU Y L, DING K N, QIU W C, et al. Discussion on molecular mechanism of Radix Bupleuri oral liquid in regulating heat stress in poultry based on network pharmacology[J]. Heilongjiang Animal Science and Veterinary Medicine, 2021(16):109-115,153-154. (in Chinese)

[14]
高娜娜, 李杨, 陈峰隆, 等. 基于网络药理学探讨枸杞多糖预防蛋雏鸡炎症性肠病的作用机制[J]. 中国兽医学报, 2025, 45(4):794-806.

GAO N N, LI Y, CHEN F L, et al. Exploring mechanism of Lycium barbarum polysaccharides in preventing inflammatory bowel disease in chicks based on network pharmacology[J]. Chinese Journal of Veterinary Science, 2025, 45(4):794-806. (in Chinese)

[15]
YANG M L, WEI H B, XU Y Z, et al. Exploring the potential mechanism of Andrographis paniculata compounds against neurodegenerative diseases based on network pharmacology and molecular docking[J]. Chinese Journal of Analytical Chemistry, 2025, 53(5):100514.

DOI

[16]
苗丁山, 杨琴, 兰燕玲, 等. 基于网络药理学与分子对接技术探讨唐草片治疗艾滋病的机制[J]. 中国艾滋病性病, 2024, 30(12):1260-1265.

MIAO D S, YANG Q, LAN Y L, et al. Exploring the mechanism of Tangcao tablet in the treatment of AIDS using network pharmacology and molecular docking techniques[J]. Chinese Journal of AIDS & STD, 2024, 30(12):1260-1265. (in Chinese)

[17]
刘佟洋, 王荣. 基于网络药理学探讨熏洗方治疗肛周脓肿术后创面愈合的作用机制[J]. 中医临床研究, 2023, 15(2):1-7.

LIU T Y, WANG R. Exploring action mechanism of fumigation prescription in the treatment of postoperative wound healing of perianal abscess based on network pharmacology[J]. Clinical Journal of Chinese Medicine, 2023, 15(2):1-7. (in Chinese)

[18]
杨熹予, 杨舒然, 张晋源, 等. 基于网络药理学及分子对接探讨青风藤治疗痛风性关节炎的作用机制[J]. 现代药物与临床, 2024, 39(5):1155-1162.

YANG X Y, YANG S R, ZHANG J Y, et al. Mechanism of Sinomenii Caulis in treatment of gout arthritis based on network pharmacology and molecular docking[J]. Drugs & Clinic, 2024, 39(5):1155-1162. (in Chinese)

[19]
王伸盛, 张新中, 张岱男, 等. 基于网络药理学、分子对接技术及实验验证探讨新橙皮苷治疗创伤性脑损伤作用机制[J]. 现代药物与临床, 2024, 39(4):857-866.

WANG S S, ZHANG X Z, ZHANG D N, et al. Mechanism of neohesperidin in treatment of traumatic brain injury based on network pharmacology,molecular docking technique and animal experiments[J]. Drugs & Clinic, 2024, 39(4):857-866. (in Chinese)

[20]
NI L, CHEN J X, WU M T, et al. Hositisines A and B,new alkaloids from the stems of Ormosia hosiei Hemsl.et Wils[J]. Natural Product Research, 2021, 35(13):2184-2189.

DOI

[21]
XU W T, LI T Z, LI S M, et al. Cytisine exerts anti-tumour effects on lung cancer cells by modulating reactive oxygen species-mediated signalling pathways[J]. Artificial Cells,Nanomedicine,and Biotechnology, 2020, 48(1):84-95.

DOI

[22]
UR RASHID H, RASOOL S, ALI Y, et al. Anti-cancer potential of sophoridine and its derivatives:recent progress and future perspectives[J]. Bioorganic Chemistry, 2020, 99:103863.

DOI

[23]
REN G, DING G T, ZHANG H Y, et al. Antiviral activity of sophoridine against enterovirus 71 in vitro[J]. Journal of Ethnopharmacology, 2019, 236:124-128.

DOI

[24]
谢莎, 王雯琪, 李文静, 等. 苦参碱药理作用、药动学及不良反应研究进展与展望[J]. 中草药, 2025, 56(10):3712-3728.

XIE S, WANG W Q, LI W J, et al. Research progress and prospect of pharmacological effects,pharmacokinetics and adverse reactions of matrine[J]. Chinese Traditional and Herbal Drugs, 2025, 56(10):3712-3728. (in Chinese)

[25]
梅文亮. 烟酸对热应激丝羽乌骨鸡生产性能及肉品质影响及机理研究[D].硕士学位论文. 南昌: 江西农业大学, 2024.

MEI W L. Effects of niacin on the production performance and meat quality of heat-stressed silkie poults and its mechanism[D].Master’s Thesis. Nanchang: Jiangxi Agricultural University, 2024. (in Chinese)

[26]
ZIMBELMAN R B, COLLIER R J, BILBY T R. Effects of utilizing rumen protected niacin on core body temperature as well as milk production and composition in lactating dairy cows during heat stress[J]. Animal Feed Science and Technology, 2013, 180(1/4):26-33.

DOI

[27]
张玉玲, 岳晓琪, 张艳丽. 苦豆子生物碱体外抑菌活性的检测[J]. 轻工科技, 2019, 35(7):33-34.

ZHANG Y L, YUE X Q, ZHANG Y L. Detection of in vitro antibacterial activity of alkaloids from Sophora alopecuroides[J]. Light Industry Science and Technology, 2019, 35(7):33-34. (in Chinese)

[28]
周微. 槐果碱在异丙肾上腺素诱导的心肌损伤中的作用及机制研究[D].博士学位论文. 南昌: 南昌大学, 2023.

ZHOU W. The role and mechanism of sophocarpine in isoproterenol-induced myocardial injury[D].Ph.D.Thesis. Nanchang: Nanchang University, 2023. (in Chinese)

[29]
陈靖, 杨丽. 槐胺碱镇痛抗炎作用及其机制研究[J]. 中华中医药学刊, 2017, 35(6):1424-1427.

CHEN J, YANG L. Study on analgesic and anti-inflammatory effects and mechanism of sophoramine[J]. Chinese Archives of Traditional Chinese Medicine, 2017, 35(6):1424-1427. (in Chinese)

[30]
贾亚杰, 齐馨馨, 魏玉, 等. 槐定碱对脂多糖诱导的肺泡上皮细胞损伤的影响[J]. 中国临床药理学杂志, 2024, 40(12):1739-1743.

JIA Y J, QI X X, WEI Y, et al. Effect of sophoridine on lipopolysaccharide-induced pulmonary alveolar epithelial cell damage[J]. The Chinese Journal of Clinical Pharmacology, 2024, 40(12):1739-1743. (in Chinese)

[31]
REVATHIDEVI S, MUNIRAJAN A K. Akt in cancer:mediator and more[J]. Seminars in Cancer Biology, 2019, 59:80-91.

DOI

[32]
LU W C, OMARI R, RAY H, et al. AKT1 mediates multiple phosphorylation events that functionally promote HSF1 activation[J]. The FEBS Journal, 2022, 289(13):3876-3893.

DOI

[33]
QIU J H, XU J B, ZHANG J X, et al. Exploring the mechanism by which BXSD decoction treats cervical cancer through the combination of network pharmacology and molecular docking analysis:molecular mechanism of AKT1 and CASP3 protein[J]. International Journal of Biological Macromolecules, 2025, 317(Pt 2):144818.

DOI

[34]
沈雨蕾, 翟烨, 郑玉敏, 等. “滋燥养荣汤”治疗表皮生长因子受体抑制剂相关皮肤干燥的回顾性研究[J]. 中日友好医院学报, 2025, 39(1):7-12.

SHEN Y L, ZHAI Y, ZHENG Y M, et al. Retrospective study on the treatment of EGFR inhibitor-related skin dryness in non-small cell lung cancer patients with “Zizao Yangrong Decoction”[J]. Journal of China-Japan Friendship Hospital, 2025, 39(1):7-12. (in Chinese)

[35]
YU J, LUAN W, YIN P, et al. Four Chinese herbal extracts prevent damage effects of heat stress in the pig small intestine via EGF/EGFR and ERK1/2 signaling pathways[J]. European Journal of Pharmacology, 2011, 668(S1):e44-e45.

[36]
BASIRICÒ L, MORERA P, PRIMI V, et al. Cellular thermotolerance is associated with heat shock protein 70.1 genetic polymorphisms in Holstein lactating cows[J]. Cell Stress and Chaperones, 2011, 16(4):441-448.

DOI

[37]
ARCHANA P R, ALEENA J, PRAGNA P, et al. Role of heat shock proteins in livestock adaptation to heat stress[J]. Journal of Dairy,Veterinary & Animal Research, 2017, 5(1):13-19.

[38]
PRODROMOU C. Mechanisms of Hsp90 regulation[J]. The Biochemical Journal, 2016, 473(16):2439-2452.

[39]
KUMAR R, KUMARI R, VERMA A, et al. Association analysis of HSP90AA1 polymorphism with thermotolerance in tropically adapted Indian crossbred cattle[J]. Tropical Animal Health and Production, 2024, 56(7):230.

DOI PMID

[40]
马轶, 冀林华, 崔森, 等. 基质金属蛋白酶9与恶性血液病关系的研究进展[J]. 医学综述, 2021, 27(8):1534-1538.

MA Y, JI L H, CUI S, et al. Research progress in relationship between matrix metalloproteinase-9 and malignant hematological diseases[J]. Medical Recapitulate, 2021, 27(8):1534-1538. (in Chinese)

[41]
GERNEZ Y, TIROUVANZIAM R, CHANEZ P. Neutrophils in chronic inflammatory airway diseases:can we target them and how?[J]. The European Respiratory Journal, 2010, 35(3):467-469.

DOI

[42]
李琦, 王岚宁, 龙欣悦, 等. 基于PI3K/Akt信号通路探讨川芎嗪对缺血性脑卒中的抗凋亡作用[J]. 中医药信息, 2025, 42(10):41-45.

LI Q, WANG L N, LONG X Y, et al. Anti-apoptotic effect of ligustrazine on ischemic stroke via PI3K/Akt signaling pathway[J]. Information on TCM, 2025, 42(10):41-45. (in Chinese)

[43]
HE S S, GUO Y H, ZHAO J X, et al. Ferulic acid protects against heat stress-induced intestinal epithelial barrier dysfunction in IEC-6 cells via the PI3K/Akt-mediated Nrf2/HO-1 signaling pathway[J]. International Journal of Hyperthermia, 2019, 35(1):112-121.

DOI PMID

[44]
DEWENTER M, VON DER LIETH A, KATUS H A, et al. Calcium signaling and transcriptional regulation in cardiomyocytes[J]. Circulation Research, 2017, 121(8):1000-1020.

DOI PMID

[45]
李樊, 李隐侠, 舒嘉傲, 等. 热应激通过钙信号调控湖羊卵巢颗粒细胞凋亡和雌激素合成[J]. 江苏农业学报, 2022, 38(5):1278-1285.

LI F, LI Y X, SHU J A, et al. Regulation of heat stress on ovarian granulosa cell apoptosis and estrogen synthesis in Hu sheep through calcium signal[J]. Jiangsu Journal of Agricultural Sciences, 2022, 38(5):1278-1285. (in Chinese)

[46]
侯昕玥, 王健全, 亢泽峰, 等. 基于UPLC-Q-TOF-MS技术及网络药理学的明视方控制近视药效物质基础与作用机制分析[J/OL]. 中国中医基础医学杂志, 2025:1-18(2025-07-03)[2025-07-08].https://doi.org/10.19945/j.cnki.issn.1006-3250.20250703.001.html.

HOU X Y, WANG J Q, KANG Z F, et al. Analysis on material basis and mechanism of Mingshi formula in controlling myopia based on UPLC-Q-TOF-MS technology and network pharmacology[J/OL]. Journal of Basic Chinese Medicine, 2025:1-18(2025-07-03)[2025-07-08].https://doi.org/10.19945/j.cnki.issn.1006-3250.20250703.001.html. (in Chinese)

[47]
许永杰, 贾晓龙, 王燕领. 茯苓酸调节cAMP/PKA/CREB信号通路对脑出血大鼠脑组织损伤的影响[J]. 安徽医药, 2025, 29(5):898-902,1061.

XU Y J, JIA X L, WANG Y L. Effect of pachymenic acid on brain tissue damage in rats with cerebral hemorrhage by regulating the cAMP/PKA/CREB signaling pathway[J]. Anhui Medical and Pharmaceutical Journal, 2025, 29(5):898-902,1061. (in Chinese)

[48]
郑秦文, 林欣, 陈思恋, 等. 热应激对体外培养小鼠乳腺上皮细胞cAMP、SOD和MDA水平的影响[J]. 黑龙江畜牧兽医, 2016(2):15-18,21,260.

ZHENG Q W, LIN X, CHEN S L, et al. Effect of heat stress on the levels of cAMP,SOD and MDA in mouse mammary epithelial cells cultured in vitro[J]. Heilongjiang Animal Science and Veterinary Medicine, 2016(2):15-18,21,260. (in Chinese)

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