Objective To investigate the potential therapeutic effects,targets,and pathways of wogonoside in hypertension-induced renal injury using the Gene Expression Omnibus(GEO)database and network pharmacology,and to validate the effects of wogonoside intervention on the renal tissues of spontaneously hypertensive rats(SHR),angiotensin Ⅱ(Ang Ⅱ)-stimulated NRK-52E cell apoptosis,and the regulation of relevant pathways through in vivo and in vitro experiments.Methods GEO dataset and network pharmacology analyses were performed to investigate the key therapeutic targets of wogonoside for hypertensive nephropathy.The STRING database was used to analyze protein-protein interactions.Biological functions were annotated via Gene Ontology(GO),and the potential signaling pathways were enriched using the Kyoto Encyclopedia of Genes and Genomes(KEGG).SHR were randomly divided into groups and given low,medium,or high doses of wogonoside(0.075,0.75,and 7.5 mg/kg)via gastric gavage for 10 weeks.Morphological changes in the kidney tissue were assessed by hematoxylin-eosin(HE)staining.Serum levels of inflammatory cytokines,including tumor necrosis factor α(TNF-α),interleukin(IL)-1 β,and IL-6,were measured using ELISA.Apoptosis rates were evaluated by TUNEL staining,and Western blot was performed to determine the expression of Bax,Bcl-2,cleaved caspase-3,and caspase-3,and the expression of phosphorylated and total extracellular signal-regulated kinases(ERK)and p38 mitogen-activated protein kinase(MAPK)proteins.An in vitro model of Ang Ⅱ-stimulated NRK-52E cells was constructed and was treated with wogonoside at different concentrations(25,50,or 100 μmol/L)for 24 h.The apoptosis rates were then assessed by Annexin V staining,and Western blot was performed to validate the expression of apoptosis-related and pathway-associated proteins.Results Analysis of dataset GSE41453 revealed 11673 upregulated and 5902 downregulated genes in the renal tissues of SHR compared to the Wistar Kyoto(WKY)rats,or the WKY control group.Through the analysis of multiple databases,371 potential targets of wogonoside were identified,resulting in 98 overlapping targets.From these,45 core therapeutic targets were identified through further analysis,including TNF,CASP3,etc.GO analysis significantly enriched processes such as the negative regulation of apoptosis.KEGG pathway enrichment analysis highlighted the apoptosis pathway,IL-17 signaling pathway,and MAPK signaling pathway as being significantly enriched.Wogonoside treatment effectively mitigated pathological damage in SHR kidney tissues and significantly inhibited the expression of inflammatory cytokines,including TNF-α,IL-1 β,and IL-6(P＜0.05).It also decreased cell apoptosis rates in SHR kidney tissues and Ang Ⅱ-stimulated NRK-52E cells,downregulated the expression of Bax and cleaved caspase-3,and upregulated Bcl-2 expression(P＜0.05).Furthermore,wogonoside treatment inhibited the phosphorylation of ERK and p38 MAPK in SHR kidney tissues and Ang Ⅱ-stimulated NRK-52E cells(P＜0.05).Conclusion Wogonoside may exert its protective effects against hypertension-induced renal injury by suppressing the inflammatory response and cell apoptosis,potentially through the regulation of the MAPK signaling pathway.

Objective To investigate the potential therapeutic effects of trifolin on hypertension-induced renal injury,as well as the key targets and pathways involved.Methods The mRNA transcriptional profiles of peripheral blood clinical samples from hypertensive patients were analyzed using Gene Expression Omnibus(GEO),a high-throughput gene expression database.The network pharmacology method was employed to screen key targets of trifolin in treating hypertension-induced renal injury.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were conducted.NRK-52E cells,a rat renal proximal tubular cell line,were used to construct an angiotensin Ⅱ(Ang Ⅱ)-stimulated cell model.Flow cytometry was performed to assess cell apoptosis rates and Western blotting was performed to determine the expression levels of apoptosis-related proteins,including Bax,Bcl-2,cleaved caspase-3,and caspase-3,and the phosphorylation and total protein levels of the key MAPK pathway proteins,including ERK,p38 MAPK,and JNK.Results Analysis of the dataset GSE75360 revealed that,compared with healthy controls,3 331 genes were upregulated and 3 197 genes were downregulated in peripheral blood mononuclear cells of hypertensive patients.According to network pharmacology analysis,472 potential targets of trifolin were identified,including CASP3 and MAPK1.Protein-protein interaction network analysis showed that these targets were closely associated with apoptosis regulatory signaling pathways.GO and KEGG pathway enrichment analyses indicated that trifolin was significantly enriched in pathways associated with negative regulation of apoptosis,apoptotic signaling pathways,and the MAPK signaling pathway.The in vitro experiments confirmed that,compared with the Ang Ⅱ group,trifolin intervention inhibited apoptosis in Ang Ⅱ-stimulated NRK-52E cells,suppressed the expression of Bax and cleaved caspase-3,promoted Bcl-2 expression,and inhibited the phosphorylation of p38 MAPK,ERK,and JNK(P<0.05).Conclusion Trifolin may exert its protective effect against hypertension-induced renal injury by inhibiting Ang Ⅱ-induced NRK-52E cell apoptosis and regulating the MAPK signaling pathway,representing an important mechanism underlying its therapeutic action.

Objective:To investigate the effects of periploca forrestii schltr in the treatment of acute gout arthritis.Methods:60 healthy male SD rats were equally randomly divided into 6 groups:normal control group( NC) ,model group( M group) ,colchicine group (C group),high doses group of periploca forrestii schltr(HD group),middle doses group of periploca forrestii schltr(MD group) and low doses group of periploca forrestii schltr( LD group).Except the normal control group,model of gouty arthritis was induced in other groups by uric acid salt,colchicine(positive control) and different dose of periploca forrestii schltr were given by intragastric ad minis-tration.Swelling dimension of joints were observed at 3,5,7 days after treatment.All rats were killed after 7 days of treatment and ankle joint tissue was taken for pathological examination and the peripheral blood of rats was prepared for detecting the expression of interleukin 1β(IL-1β),IL-6,IL-8 and tumor necrosis factor(TNF-α) using enzyme linked immunosorbent test(ELISA).Results:The ankle joint swelling of periploca forrestii schltr group was significantly lower than that in the model group,and the effect of high doses group was much better than the low doses group after 7 days treatment(P<0.05);compared with model group,the inflammatory cells of each treatment groups were decreased and high doses group did not differ from that of normal control group;the levels of IL-1β,IL-6,IL-8 and TNF-αin periploca forrestii schltr group were dramatically lower than those in the model group in a dose-dependent manner.Conclusion:Periploca forrestii schltr has good therapeutic effect in rats with acute gouty arthritis and shows a dose-dependent response,and the mechanism may relate to the inhibition of inflammatory cytokines expression.

Objective To investigate the immunomodulatory effects of Tetrastigma hypoglaucum Planch in the treatment of rheumatoid arthritis. Methods Sixty SD rats were ramdomly divided into normal control group, model control group,low-,middle-and high-dose Tetrastigma hypoglaucum Planch groups (50,100 and 200 mg.kg-1,respectively),and tripterygium glycosides tablet group. Except for normal control group,rheumatoid arthritis model was established by using bovine typeⅡcollagen in SD rats of the other groups. Organ index,plasma levels of IL-1β,IL-6,TNF-αand anti-CⅡin rat rheumatoid arthritis models were evaluated. Results The kidney index of the low-dose/middle-dose Tetrastigma hypoglaucum Planch groups were significantly different from that of the model control group (P<0.05). Significant difference was also found in the spleen index between the high-dose Tetrastigma hypoglaucum Planch group and the normal control group ( P<0.05) . The plasma levels of IL-1β,IL-6,TNF-αand anti-CⅡantibody were significantly higher in the model control group than in the normal control group. Meanwhile, the plasma IL-1β, IL-6 and TNF-α levels of the middle-dose/high-dose Tetrastigma hypoglaucum Planch groups were significantly lower than those of the model group ( P<0.01) . The levels of anti-CII antibody were significantly lower in the low-,middle-and high-dose Tetrastigma hypoglaucum Planch groups than in the model control group. Conclusion It is worthwhile to explore and develop Tetrastigma hypoglaucum Planch since it possesses immunomodulatory effects and may be applied in the treatment of rheumatoid arthritis.