基于IKKβ/NF-κB信号通路探讨藏红花素减轻高糖诱导肾小球内皮细胞损伤的作用机制

doi:10.3969/j.issn.1006-5725.2026.12.016

Abstract

Abstract:

Objective To investigate the mechanism by which crocin alleviates high glucose-induced glomerular endothelial cell damage based on the inhibitor of nuclear factor kappa B kinase β （IKKβ）/nuclear transcription factor-κB （NF-κB） signaling pathway. Methods High glucose-induced human glomerular endothelial cells were treated with crocin at concentrations of 0， 12.5， 25， 50， and 75 μmol/L， and the optimal concentration of crocin was screened via the CCK-8 experiment. Glomerular endothelial cells were randomly assigned to the normal group， osmotic pressure control group， high glucose group， crocin group， phorbol ester （PMA） group， and crocin + PMA group. Except for the normal group and osmotic pressure control group， all the other groups were induced with high glucose and treated with crocin and the IKKβ/NF-κB signaling pathway activator PMA according to different groups for 48 hours. Immunofluorescence staining was employed to observe the cytoskeleton of the cells. The CCK-8 experiment， Transwell experiment， and TdT-mediated dUTP nick end labeling （TUNEL） staining were carried out to detect cell viability， permeability， and apoptosis， respectively. The oxidative stress and inflammation-related factors in the cells were measured. Western blot （WB） was utilized to detect the expression of proteins related to apoptosis， tight junctions， and the IKKβ/NF-κB signaling pathway. Results Compared with the normal group and the osmotic pressure control group， the high-glucose group exhibited a decrease in the activity of glomerular endothelial cells， the activities of superoxide dismutase （SOD） and glutathione peroxidase （GSH-Px）， the relative expression of occludin protein， and the relative expressions of zonula occludens-1 （ZO-1） and Bcl-2 protein （P < 0.05）. Meanwhile， it showed an increase in permeability， apoptosis rate， malondialdehyde （MDA） content， interleukin （IL）-6 and IL-1β levels， the relative expression of Bax protein， and the ratios of p-IKKβ/IKKβ， p-IκB-α/IκB-α， and p-NF-κB p65/NF-κB p65 （P < 0.05）. Compared with the high-glucose group， the crocin group showed an increase in the activity of glomerular endothelial cells， the activities of SOD and GSH-Px， the relative expression of occludin protein， and the relative expressions of ZO-1 and Bcl-2 protein （P < 0.05）. In addition， it had a decrease in permeability， apoptosis rate， MDA content， IL-6 and IL-1β levels， the relative expression of Bax protein， and the ratios of p-IKKβ/IKKβ， p-IκB-α/IκB-α， and p-NF-κB p65/NF-κB p65 （P < 0.05）. The trend of changes in various indicators in the PMA group was opposite to that in the crocin group （P < 0.05）. Compared with the crocin group， the crocin + PMA group showed a decrease in the activity of glomerular endothelial cells， the activities of SOD and GSH-Px， the relative expression of occludin protein， and the relative expressions of ZO-1 and Bcl-2 protein （P < 0.05）. Moreover， it presented an increase in permeability， apoptosis rate， MDA content， IL-6 and IL-1β levels， the relative expression of Bax protein， and the ratios of p-IKKβ/IKKβ， p-IκB-α/IκB-α， and p-NF-κB p65/NF-κB p65 （P < 0.05）. Conclusion Crocin alleviates the damage to glomerular endothelial cells induced by high glucose by inhibiting the activation of the IKKβ/NF-κB signaling pathway.

Key words: crocin, IKKβ/NF-κB, high glucose, glomerular endothelial cells, damage

CLC Number:

R587.2

Hongyun WANG,Chengcai ZHANG,Weiwei WANG. Exploring the mechanism of crocin in alleviating high glucose induced glomerular endothelial cell damage based on the IKKβ/NF⁃κB signaling pathway[J]. The Journal of Practical Medicine, 2026, 42(12): 2208-2215.

Figures/Tables 9

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References 27

[1]	高晶，赵锦，王凡，等. 不同频率血液透析滤过联合高通量血液透析治疗对糖尿病肾病维持性血液透析患者肾功能，微炎症状态及左心功能的影响［J］. 实用医学杂志， 2025， 41（21）： 3398-3404. doi： 10.3969/j.issn.1006-5725.2025.21.015 . doi: 10.3969/j.issn.1006-5725.2025.21.015
[2]	LUO Y， ZHANG W， QIN G. Metabolomics in diabetic nephropathy： Unveiling novel biomarkers for diagnosis （Review）［J］. Mol Med Rep， 2024， 30（3）： 156-168. doi： 10.3892/mmr.2024. 13280 . doi: 10.3892/mmr.2024. 13280
[3]	WANG G， ZHAO J， ZHOU M，et al. Unveiling diabetic nephropathy： A novel diagnostic model through single-cell sequencing and co-expression analysis［J］. Aging （Albany NY）， 2024， 16（13）： 10972-10984. doi： 10.18632/aging.205982 . doi: 10.18632/aging.205982
[4]	ZHOU J， FRANCESCHINI N， TOWNLEY-TILSON W H D， et al. Nutritional Strategies against Diabetic Nephropathy： Insights from Animal Studies and Human Trials［J］. Nutrients， 2024， 16（12）： 1918-1932. doi： 10.3390/nu16121918 . doi: 10.3390/nu16121918
[5]	LI Y， ZHANG J， ZHU Y. METTL14 derived from exosomes of M1 macrophages promotes high glucose-induced apoptosis， inflammation and oxidative stress in glomerular endothelial cells by mediating PAQR3 m6A modification［J］. Clin Exp Nephrol， 2024， 28（12）： 1221-1231. doi： 10.1007/s10157-024-02536-0 . doi: 10.1007/s10157-024-02536-0
[6]	XUE H Z， CHEN Y， WANG S D，et al. Radix Astragali and Its Representative Extracts for Diabetic Nephropathy： Efficacy and Molecular Mechanism［J］. J Diabetes Res， 2024， 2024： 5216113-5216138. doi： 10.1155/2024/5216113 . doi: 10.1155/2024/5216113
[7]	WANG X， LIU M， LI X，et al. Utilizing molecular docking and cell validation to explore the potential mechanisms of lupenone attenuating the inflammatory response via NF-κB pathway［J］. Sci Rep， 2024， 14（1）： 625-634. doi：10.1038/s41598-024- 51150-3 . doi: 10.1038/s41598-024- 51150-3
[8]	WEI T T， YANG L T， GUO F，et al. Activation of GPR120 in podocytes ameliorates kidney fibrosis and inflammation in diabetic nephropathy［J］. Acta Pharmacol Sin， 2021， 42（2）： 252-263. doi： 10.1038/s41401-020-00520-4 . doi: 10.1038/s41401-020-00520-4
[9]	LOTFY M， KHALAF A， ALBARGHOUTHY F，et al. Effects of Conocarpus lancifolius and crocin on superoxide dismutase and protein kinase B genes and protein expressions in diabetic rats［J］. PLoS One， 2025， 20（6）： e0326676-e0326694. doi： 10.1371/journal.pone.0326676 . doi: 10.1371/journal.pone.0326676
[10]	MAFI A， MOKHTARI Z， HOSSEINI E，et al. Effect of Saffron （Crocus sativus） Supplementation on Oxidative Stress， Inflammatory Indices， and Renal and Liver Function Parameters in Patients With Type 2 Diabetes Mellitus： A GRADE-Assessed Systematic Review and Meta-analysis of Randomized Clinical Trials［J］. Nutr Rev， 2025， 83（6）： 971-987. doi： 10.1093/nutrit/nuae121 . doi: 10.1093/nutrit/nuae121
[11]	JAAFARINIA A， KAFAMI B， SAHEBNASAGH A，et al. Evaluation of therapeutic effects of crocin in attenuating the progression of diabetic nephropathy： A preliminary randomized triple-blind placebo-controlled trial［J］. BMC Complement Med Ther， 2022， 22（1）： 262-270. doi： 10.1186/s12906-022-03744-5 . doi: 10.1186/s12906-022-03744-5
[12]	ZHANG J， ZHAO X， ZHU H，et al. Crocin protects the renal tubular epithelial cells against high glucose-induced injury and oxidative stress via regulation of the SIRT1/Nrf2 pathway［J］. Iran J Basic Med Sci， 2022， 25（2）： 193-197. doi： 10.22038/IJBMS.2022. 51597.11708 . doi: 10.22038/IJBMS.2022. 51597.11708
[13]	HASSAN M H， SALAMA S A， ISMAIL R S. Targeting NF-kappa B/proinflammatory cytokines/TGF-β/TIMP-1 pathway by crocin enhances recovery from hepatic fibrosis in rats［J］. Sci Rep， 2025， 16（1）： 2072-2081. doi： 10.1038/s41598-025-32672-w . doi: 10.1038/s41598-025-32672-w
[14]	王延海，张雷明，冯艳艳. 大黄素改善高糖条件中人肾小球血管内皮细胞炎症，氧化应激及凋亡作用的研究［J］. 中国临床药理学杂志， 2023， 39（10）： 1422-1426. doi： 10.13699/j.cnki. 1001-6821.2023.10.012 . doi: 10.13699/j.cnki. 1001-6821.2023.10.012
[15]	CHEN M， LIN X， ZHANG L，et al. Effects of nuclear factor-κB signaling pathway on periodontal ligament stem cells under lipopolysaccharide-induced inflammation［J］. Bioengineered， 2022， 13（3）： 7951-7961. doi： 10.1080/21655979.2022. 2051690 . doi: 10.1080/21655979.2022. 2051690
[16]	WU X Q， ZHAO L， ZHAO Y L，et al. Traditional Chinese medicine improved diabetic kidney disease through targeting gut microbiota［J］. Pharm Biol， 2024， 62（1）： 423-435. doi： 10.1080/13880209.2024.2351946 . doi: 10.1080/13880209.2024.2351946
[17]	HUSHMANDI K， KLIONSKY D J， FARAHANI N， et al. Regulation of pyroptosis in diabetic nephropathy by long non-coding and circular RNAs［J］. Clin Exp Med， 2025， 25（1）： 208-239. doi： 10.1007/s10238-025-01740-w . doi: 10.1007/s10238-025-01740-w
[18]	MŁYNARSKA E， BUŁAWSKA D， CZARNIK W，et al. Novel Insights into Diabetic Kidney Disease［J］. Int J Mol Sci， 2024， 25（18）： 10222-10245. doi： 10.3390/ijms251810222 . doi: 10.3390/ijms251810222
[19]	HUANG M， CHANG J， LIU Y， et al. Apelin/APJ alleviates diabetic nephropathy by improving glomerular endothelial cells dysfunction via SIRT3‑KLF15［J］. Mol Med Rep， 2025， 31（5）： 122-133. doi： 10.3892/mmr.2025.13487 . doi: 10.3892/mmr.2025.13487
[20]	YANG Y， WANG Y， ZHOU Y，et al. Tirzepatide alleviates oxidative stress and inflammation in diabetic nephropathy via IL-17 signaling pathway［J］. Mol Cell Biochem，2025， 480（2）： 1241-1254. doi： 10.1007/s11010-024-05066-1 . doi: 10.1007/s11010-024-05066-1
[21]	GHAVIDEL F， AMIRI H， TABRIZI M H， et al. The Combinational Effect of Inulin and Resveratrol on the Oxidative Stress and Inflammation Level in a Rat Model of Diabetic Nephropathy［J］. Curr Dev Nutr， 2023， 8（1）： 102059-102070. doi： 10.1016/j.cdnut.2023.102059 . doi: 10.1016/j.cdnut.2023.102059
[22]	DASTAN M， RAJAEI Z， SHARIFI M， et al. Crocin Improves Cognitive Impairment in LPS-treated Rats through Anti-Apoptotic， Anti-Inflammatory， and Antioxidant Activities［J］. Mol Neurobiol， 2025， 62（5）： 5804-5815. doi： 10.1007/s12035-024-04638-y . doi: 10.1007/s12035-024-04638-y
[23]	ZIANI A， BEKKOUCH O， OUAHHOUD S，et al. Phytochemistry， Biological Activities， Molecular Mechanisms， and Toxicity of Saffron （Crocus sativus L.）： A Comprehensive Overview［J］. Antioxidants （Basel）， 2025， 14（12）： 1433-1478. doi： 10.3390/antiox14121433 . doi: 10.3390/antiox14121433
[24]	CHEN W， SU J， LIU Y，et al. Crocin Ameliorates Diabetic Nephropathy through Regulating Metabolism， CYP4A11/PPARγ， and TGF-β/Smad Pathways in Mice［J］. Curr Drug Metab， 2023， 24（10）： 709-722. doi： 10.2174/011389200225792823103111 3337 . doi: 10.2174/011389200225792823103111 3337
[25]	MOHAMMADI Y， ZANGOOEI M， ZARDAST M， et al. The effect of crocin and losartan on TGF-β gene expression and histopathology of kidney tissue in a rat model of diabetic nephropathy［J］. Avicenna J Phytomed， 2023， 13（2）： 189-199. doi： 10. 22038/AJP.2022.21414 . doi: 10. 22038/AJP.2022.21414
[26]	ZHANG R， QIN L， SHI J. MicroRNA‑199a‑3p suppresses high glucose‑induced apoptosis and inflammation by regulating the IKKβ/NF‑κB signaling pathway in renal tubular epithelial cells［J］. Int J Mol Med， 2020， 46（6）： 2161-2171. doi： 10.3892/ijmm.2020.4751 . doi: 10.3892/ijmm.2020.4751
[27]	WANG Y， LI H， JIANG S，et al. The glycolytic enzyme PFKFB3 drives kidney fibrosis through promoting histone lactylation-mediated NF-κB family activation［J］. Kidney Int， 2024， 106（2）： 226-240. doi： 10.1016/j.kint.2024.04.016 . doi: 10.1016/j.kint.2024.04.016

藏红花素浓度	肾小球内皮细胞活性
藏红花素浓度	高糖诱导	正常培养
0 μmol/L	100.00 ± 0.00	100.00 ± 0.00
12.5 μmol/L	140.98 ± 12.15^*	103.12 ± 11.45
25 μmol/L	177.13 ± 13.64^*	105.47 ± 13.20
50 μmol/L	221.86 ± 14.57^*	104.83 ± 14.04
100 μmol/L	240.12 ± 15.38^*	102.94 ± 15.18
F值	126.692	0.185
P值	< 0.001	0.944

组别	细胞活性	通透性	凋亡率
正常组	100.00 ± 0.00	100.00 ± 0.00	4.02 ± 1.21
渗透压对照组	60.83 ± 6.15^*	238.03 ± 26.34^*	42.01 ± 3.64^*
高糖组	61.12 ± 6.31^*	237.12 ± 25.83^*	41.90 ± 3.53^*
藏红花素组	92.13 ± 4.76^&	129.35 ± 30.46^&	8.95 ± 2.43^&
PMA组	31.05 ± 4.92^&	396.84 ± 29.12^&	70.76 ± 4.18^&
藏红花素+PMA组	64.01 ± 5.95^#	218.27 ± 27.05^#	38.65 ± 4.01^#
F值	138.953	101.456	326.861
P值	< 0.001	< 0.001	< 0.001

组别	SOD/（U/mg prot）	GSH-Px/（U/mg prot）	MDA/（nmol/mg prot）	IL-6/（pg/mL）	IL-1β/（pg/mL）
正常组	74.31 ± 5.92	14.72 ± 1.38	7.28 ± 1.24	63.54 ± 11.16	95.43 ± 18.21
渗透压对照组	29.75 ± 4.87^*	6.92 ± 0.86^*	13.94 ± 2.52^*	245.12 ± 20.35	305.85 ± 26.42^*
高糖组	30.12 ± 4.53^*	6.81 ± 0.70^*	14.03 ± 2.07^*	242.83 ± 17.32^*	307.56 ± 23.14^*
藏红花素组	69.84 ± 5.47^&	13.04 ± 1.31^&	8.30 ± 1.52^&	73.60 ± 13.41^&	110.87 ± 17.53^&
PMA组	8.45 ± 2.31^&	1.59 ± 0.47^&	20.74 ± 2.19^&	415.95 ± 19.15^&	512.35 ± 24.60^&
藏红花素+PMA组	33.67 ± 4.26^#	7.46 ± 0.71^#	12.86 ± 1.92^#	231.74 ± 20.23^#	291.20 ± 21.83^#
F值	179.037	146.512	36.543	341.964	286.807
P值	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001

组别	occludin	ZO-1	Bcl-2	Bax
正常组	0.97 ± 0.12	1.10 ± 0.13	0.91 ± 0.09	0.23 ± 0.04
渗透压对照组	0.54 ± 0.10^*	0.70 ± 0.11^*	0.48 ± 0.07^*	0.64 ± 0.08^*
高糖组	0.56 ± 0.08^*	0.69 ± 0.10^*	0.49 ± 0.05^*	0.65 ± 0.06^*
藏红花素组	0.94 ± 0.10^&	1.07 ± 0.11^&	0.89 ± 0.07^&	0.24 ± 0.05^&
PMA组	0.17 ± 0.05^&	0.29 ± 0.07^&	0.10 ± 0.03^&	1.09 ± 0.10^&
藏红花素+PMA组	0.58 ± 0.09^#	0.71 ± 0.12^#	0.51 ± 0.06^#	0.63 ± 0.08^#
F值	61.457	45.368	132.067	120.205
P值	< 0.001	< 0.001	< 0.001	< 0.001

组别	p-IKKβ/IKKβ	p-IκB-α/IκB-α	p-NF-κB p65/NF-κB p65
正常组	0.11 ± 0.02	0.09 ± 0.01	0.12 ± 0.02
渗透压对照组	0.52 ± 0.08^*	0.47 ± 0.06^*	0.51 ± 0.09^*
高糖组	0.51 ± 0.06^*	0.48 ± 0.05^*	0.50 ± 0.06^*
藏红花素组	0.12 ± 0.03^&	0.10 ± 0.02^&	0.13 ± 0.03^&
PMA组	0.93 ± 0.05^&	0.91 ± 0.06^&	0.94 ± 0.04^&
藏红花素+PMA组	0.49 ± 0.07^#	0.47 ± 0.04^#	0.48 ± 0.07^#
F值	179.012	280.678	169.231
P值	< 0.001	< 0.001	< 0.001

Exploring the mechanism of crocin in alleviating high glucose induced glomerular endothelial cell damage based on the IKKβ/NF⁃κB signaling pathway

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