Research On The Anti-Ovarian Cancer Mechanism Of Aconitum Soongaricum Stapf. Processed Products Based On "Effect-Toxicity" Network Analysis And In Vivo Experiments

Objective:To study the mechanism of anti-ovarian cancer of Aconitum soongaricum Stapf.concocted products by“effect-toxicity”network analysis and in vivo experiments.Methods 1.The“effect-toxicity”components and targets of Aconitum soongaricum Stapf.products were searched through databases and literature.Complete the induction of potential targets of“effect-toxicity”.Mapping of the Aconitum soongaricum Stapf.-target relationship network.R software for gene function annotation analysis.2.a.Construction of SKOV-3 tumor-bearing nude mouse model:9 groups were set up in the experiment,the tumor size of each group was compared,and the tumor suppression rate was calculated.b.HE staining,observation of pathological tissue sections.c.ELISA detected and compared the expression of IL-6,IL-1β,TNF-αinflammatory factors in serum of each group.d.q RT-PCR and IHC detected and compared the relative expressions of PIK3CA,AKT1,E-cadherin,and N-cadherin genes and proteins.e.WB detected and compared the relative expression of proteins in PIK3CA,p-PIK3CA,AKT1,p-AKT1,E-cadherin,and N-cadherin.3.Cardiomyocyte H9c2 experiment,CCK-8 method to calculate IC₅₀,flow cytometry to detect apoptosis.Results:1.3 components of Aconitum soongaricum Stapf.-ovarian cancer and 100 key targets were retrieved,which mainly exerted the role of ovarian cancer by mediating the PI3K/Akt and Fox O signaling pathway.2.In vivo experimental verification:a.Compared with the model group,songorine,aconitine and benzoylaconine can significantly inhibit tumor growth（P<0.05）.b.ELISA results showed that the expression of serum inflammatory factors IL-6,IL-1βand TNF-αin the songorine,aconitine and benzoylaconine groups was significantly reduced compared with the model group（P<0.05）.Compared with the PI3K inhibitor（LY294002）group,the expression of serum inflammatory factors IL-6,IL-1βand TNF-αin the drug+LY294002 group was significantly reduced（P<0.05）.c.The results of tissue staining of HE and IHC sections showed that tumor cells had the greatest degree and extent of tumor cell necrosis in the benzoylaconitine group.d.q RT-PCRresults showed that compared with the model group,the expression of E-cadherin m RNA in the songorine,aconitine and benzoylaconine groups was significantly increased（P<0.05）,the expression of N-cadherin m RNA was significantly reduced（P<0.05）,and the expression of PIK3CA and AKT1 m RNA was reduced,but there was no significant difference.Compared with LY294002 group,the expression of AKT1 and N-cadherin m RNA in the drug+LY294002 group was significantly reduced（P<0.05）,and the expression of E-cadherin m RNA in the songorine+LY294002 group and aconitine+LY294002 group was significantly increased（P<0.05）.e.The results of WB showed that compared with the model group,the expression of E-cadherin protein in the songorine,aconitine and benzoylaconine groups was significantly increased（P<0.05）,the expression of N-cadherin,p-PIK3CA and p-AKT1 proteins in the songorine and aconitine groups was significantly reduced（P<0.05）,and the expression of p-PIK3CAand p-AKT1 proteins in the aconitine group was significantly reduced（P<0.05）.Compared with LY294002 group,the expression of E-cadherin protein in the drug+LY294002 group was significantly increased,and the expression of N-cadherin,p-PIK3CA and p-AKT1 protein was significantly reduced（P<0.05）.3.a.18 toxic components of Aconitum soongaricum Stapf.and 22 Aconitum soongaricum Stapf.-cardiotoxic targets were excavated through network toxicology.It mainly induces cardiotoxic effects by mediating the MAPK,IL-17 and Fox O signaling pathway.b.The results of cardiomyocyte experiments showed that the IC₅₀of aconitine,songorine and benzoylaconitine respectively were 0.357,0.617 and 1.070mg/m L,and the apoptosis rate were:aconitine>songorine>benzoylaconitine,the content of benzoylaconine in Aconitum soongaricum Stapf.products was 0.745%,which was the main hydrolysate product of aconitine,and the myocardial toxicity of benzoylaconitine was about 1/3 of aconitine.Conclusions:1.The“effect-poison”network combined with biological experiments showed that the anti-ovarian cancer efficacy components of Xinjiang characteristic medicinal herb-Aconitum soongaricum Stapf.were mainly aconitine,songorine and benzoylaconitine,the target may be PIK3CA,AKT1,AKT2,etc.and the action pathway may be PI3K/Akt,Ras,Fox O signaling pathway,etc.2.The results of pharmacodynamic experiments confirmed that the main alkaloids of Aconitum soongaricum Stapf.products can inhibit thegrowth of nude mouse ovarian cancer tumors,affect the transformation of inflammatory factors IL-6,IL-1β,TNF-αand epithelial-mesenchymal N-cadherin,E-cadherin and p-PIK3CA and p-AKT1 proteins,and the mechanism may be related to the PI3K/AKT signaling pathway.3.The toxic component of Aconitum soongaricum Stapf.is aconitine,which is mostly hydrolyzed to benzoylaconitine after processing,and the toxicity of cardiomyocytes is reduced,and the attenuation mechanism may be related to the regulation of plasminogen（PLG）targets.