OBJECTIVE:To explore the medication patterns and mechanisms of action of Shaofu Zhuyu decoction(少腹逐瘀汤,SFZYD)in inhibiting ferroptosis through the nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase 1...OBJECTIVE:To explore the medication patterns and mechanisms of action of Shaofu Zhuyu decoction(少腹逐瘀汤,SFZYD)in inhibiting ferroptosis through the nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase 1(HO-1)/glutathione peroxidase 4(GPX4)pathway to improve diabetes mellitus-induced erectile dysfunction(DMED).METHODS:Firstly,data mining was employed to identify the medication patterns of Traditional Chinese Medicine(TCM)in treating DMED.Secondly,network pharmacology combined with a ferroptosis database was used to predict the targets.Subsequently,cell counting kit-8,4',6-diamidino-2-phenylindole staining,reverse transcription-polymerase chain reaction(RT-PCR),and reagent kits were utilized to assess the repair effects of SFZYD on corpus cavernosum endothelial cells(CCECs)induced by high glucose(HG).Metabolic indicators,hematoxylin-eosin staining,and Masson staining were performed to observe the restorative effects of SFZYD on erectile function and penile tissue in diabetic rats.Finally,using Nrf2 inhibitors,the expression of related proteins and mRNAs was detected through Western blotting and RT-PCR.Reactive oxygen species levels and mitochondrial membrane potential were detected by flow cytometry.RESULTS:Data mining revealed that the prescription rules for blood stasis-type DMED coincide with the treatment principles of SFZYD.Network pharmacology identified 48 ferroptosis-related targets,primarily heme oxygenase 1(HMOX1)and GPX4.Kyoto Encyclopedia of Genes and Genomes enrichment analysis associated these targets with the ferroptosis pathway.SFZYD repaired HG-induced CCECs damage and restored HMOX1 and GPX4 mRNA levels.in vivo,SFZYD effectively alleviated erectile dysfunction and repaired blood sinuses and fibrosis in diabetic rats.Following Nrf2 inhibition,the expression of Nrf2,HMOX1,and GPX4 decreased,while SFZYD intervention reversed these effects,improving ferroptosis and oxidative stress indicators.CONCLUSION:This study explored the potential mechanisms and efficacy of the TCM prescription SFZYD in treating DMED through data mining,network pharmacology analysis,cellular experiments,and animal experiments.It verified its effectiveness in repairing HGinduced CCECs damage,improving the pathological state of penile tissue in diabetic rats,and restoring erectile function by regulating the Nrf2/HO-1/GPX4 signaling pathway.This provides new insights and scientific evidence for treating DMED with TCM.展开更多
基金National Natural Science Foundation of China(Mechanism of Ginsenoside Rk1 in Treating Ulcerative Colitis by Regulating Macrophage Polarization Based on Epidermal Growth Factor Receptor Phosphorylation,No.82374128)Department of Science and Technology of Jilin Province(Research on the Mechanism of Endogenous Stem Cells in Mechanical Force Therapy for Neurogenic Erectile Dysfunction,No.ydzj202301zyts138)+1 种基金Projects in Health Commission of Jilin Province(Research on the Mechanism of Low-intensity Pulsed Ultrasound in the Treatment of Late-onset Hypogonadism via the Wnt Family Member 5a/betaCatenin Axis,No.2022jc043Research on the Mechanism of Microencapsulated Adipose-Derived Stem Cells in the Treatment of Neurogenic Erectile Dysfunction,No.2022jc048)。
摘要OBJECTIVE:To explore the medication patterns and mechanisms of action of Shaofu Zhuyu decoction(少腹逐瘀汤,SFZYD)in inhibiting ferroptosis through the nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase 1(HO-1)/glutathione peroxidase 4(GPX4)pathway to improve diabetes mellitus-induced erectile dysfunction(DMED).METHODS:Firstly,data mining was employed to identify the medication patterns of Traditional Chinese Medicine(TCM)in treating DMED.Secondly,network pharmacology combined with a ferroptosis database was used to predict the targets.Subsequently,cell counting kit-8,4',6-diamidino-2-phenylindole staining,reverse transcription-polymerase chain reaction(RT-PCR),and reagent kits were utilized to assess the repair effects of SFZYD on corpus cavernosum endothelial cells(CCECs)induced by high glucose(HG).Metabolic indicators,hematoxylin-eosin staining,and Masson staining were performed to observe the restorative effects of SFZYD on erectile function and penile tissue in diabetic rats.Finally,using Nrf2 inhibitors,the expression of related proteins and mRNAs was detected through Western blotting and RT-PCR.Reactive oxygen species levels and mitochondrial membrane potential were detected by flow cytometry.RESULTS:Data mining revealed that the prescription rules for blood stasis-type DMED coincide with the treatment principles of SFZYD.Network pharmacology identified 48 ferroptosis-related targets,primarily heme oxygenase 1(HMOX1)and GPX4.Kyoto Encyclopedia of Genes and Genomes enrichment analysis associated these targets with the ferroptosis pathway.SFZYD repaired HG-induced CCECs damage and restored HMOX1 and GPX4 mRNA levels.in vivo,SFZYD effectively alleviated erectile dysfunction and repaired blood sinuses and fibrosis in diabetic rats.Following Nrf2 inhibition,the expression of Nrf2,HMOX1,and GPX4 decreased,while SFZYD intervention reversed these effects,improving ferroptosis and oxidative stress indicators.CONCLUSION:This study explored the potential mechanisms and efficacy of the TCM prescription SFZYD in treating DMED through data mining,network pharmacology analysis,cellular experiments,and animal experiments.It verified its effectiveness in repairing HGinduced CCECs damage,improving the pathological state of penile tissue in diabetic rats,and restoring erectile function by regulating the Nrf2/HO-1/GPX4 signaling pathway.This provides new insights and scientific evidence for treating DMED with TCM.