Procyanidin B2 Inhibits Cisplatin-induced DNA Damage And Inflammatory Responses By Regulating Sirt1/Sirt3

Objective:This article aims to study the effects of procyanidin B2(PCB2)on DNA damage and secretion of inflammatory factors in THP-1 cells induced by cisplatin,and to further explore the Sirt1/Sirt3 inflammation-related pathways and mitochondrial damage pathways PCB2 inhibits the possible mechanism of cisplatin-induced DNA damage and regulates the secretion of inflammatory factors,and then clarifies the specific mechanism of Sirt1/Sirt3 inflammation-related pathways and mitochondrial damage pathways,providing a basis for the clinical application of PCB2.Methods:1.Using Trypan blue staining method,inoculate THP-1 cells on a cell culture plate,stain dead cells,count the number of cells including live cells,and detect cell viability.2.Using the MTT colorimetric method,inoculate cells to the cell culture plate,set as the control group,the cisplatin treatment group and the PCB2 treatment group,calculate their ratios to obtain the relative survival rate,and screen out the more suitable procyanidin B2 action time And drug concentration..3.After the cells are inoculated and infected,single-cell gel electrophoresis(comet test)is used to detect DNA damage,and Western Blot and cell immunofluorescence are used to detect the change of ?H2AX.4.The cytokine concentration in the cell culture supernatant was determined by the ELISA method.5.Use the DCFH-DA probe method to detect the level of ROS in THP-1 cells after PCB2 intervention.6.Perform real-time fluorescent quantitative PCR experiments on THP-1 cells after PCB2 intervention,and find the expression levels of IL-1?,IL-6 and TNF-?m RNA in THP-1 cells.7.Western Blot was used to detect the expression levels of Sirt1,Sirt3,and NF-?Bp65 protein in THP-1 cells after the intervention of proanthocyanidins.Results:1.Compared with the control group,a certain concentration of cisplatin can induce cell damage,and PCB2 can improve DNA damage to a certain extent,which is concentration-dependent and has a significant difference(*P<0.05).2.Compared with the control group,the levels of inflammatory cytokines IL-1?,IL-6 and TNF-? in the cells treated with cisplatin were significantly increased,with significant differences(*P<0.05).3.Cisplatin treatment can induce the release of pro-inflammatory cytokines(IL-1?,IL-6 and TNF-?);PCB2 can reduce the release of IL-1?,IL-6 and TNF-?.Compared with the control group,#P<0.05;compared with the cisplatin treatment,*P<0.05.4.PCB2 can reduce the activity of Sirt/Sirt3 related signaling pathways and the activity of mitochondria,thereby affecting the secretion of inflammatory factors.Conclusion:PCB2 can inhibit cisplatin-induced DNA damage and inflammation.The possible mechanism is to mediate the Sirt1/Sirt3 pathway and the mitochondrial damage pathway.The above studies indicate that PCB2 may be developed as a routine by inhibiting inflammation-related pathways.Adjuvant for anti-inflammatory drugs.