Effects And Mechanism Of Ropivacaine On Inhibition Of Proliferation And Apoptosis Of Sk-hep-1 Cells

Background:Recent studies have shown that local anesthetics play an anti-tumor role through different mechanisms.Ropivacaine is a new type of long-acting amide local anesthetics,which is generally used for local infiltration anesthesia,intraspinal anesthesia,regional nerve block,especially for local anesthesia and postoperative analgesia.With the in-depth study of the anti-tumor effect of local anesthetics,the anti-tumor effect of ropivacaine has attracted more and more scholars’interest.Objective:To observe the effect of ropivacaine on the proliferation and apoptosis of human hepatocellular carcinoma Sk-hep-1 cells and explore its anti-tumor mechanism.Methods:The human hepatocellular carcinoma Sk-hep-1 cell line was cultured in vitro.The cell viability of Sk-hep-1 cell line and human normal hepatocyte L-02 cell line were observed 24 hours after treatment with 0 μM,400 μM,800 μM and 1200μM Ropivacaine by MTT.The cell viability of Sk-hep-1 cell line was observed 24 hours,48 hours and 72 hours after treatment with 400 μM,800 μM and 1200 μM Ropivacaine.Morphological changes of human hepatocellular carcinoma Sk-hep-1 cells were observed by DAPI nuclear staining after different concentrations of Ropivacaine(400 μM,800 μM,1200μM)treated for 24 hours.After treatment of Sk-hep-1 cells with 400 μM,800 μM and 1200 μM Ropivacaine for 24 hours,the effects of ropivacaine on the expression of apoptotic-related proteins such as Cleaved Caspase-3,Cleaved Caspase-9,Bcl-2,Bax and Cyt-c were detected by Western blotting.Results:MTT assay showed that 400 μM,800 μM and 1200 μM Ropivacaine had no cytotoxic effect and had no significant effect on the survival rate of normal hepatocytes(P>0.05).Compared with the same group,400 μM,800 μM and 1200 μM Ropivacaine could significantly inhibit the cell viability of human hepatocellular carcinoma Sk-hep-1 cells 24,48 and 72 hours after treatment(P<0.05,P<0.01,P<0.01),and the IC50 value of ropivacaine at 24 hours was 1303 μM,,but there was no significant difference at the same concentration at different time points(P>0.05).Compared with the 0-mu-M group,after 24 hours of intervention with 400μM,800 μM and 1200 μM ropivacaine,the nuclei of human hepatocellular carcinoma Sk-hep-1 cells showed atrophy,fragmentation and apoptotic morphological changes such as nuclear edge collection.After treated with ropivacaine at different concentrations for 24 hours,the expression levels of apoptotic related factors Bax,Cleaved caspase-3,Cleaved Caspase-9 and Cyt-c were significantly increased(P<0.01),while the expression levels of apoptotic inhibitor protein Bcl-2 were significantly decreased(P<0.01),showing a concentration-dependent relationship.Conclusion:1.Ropivacaine has a significant inhibitory effect on the growth of human hepatocellular carcinoma cell line Sk-hep-1,which is related to the apoptosis of hepatocellular carcinoma cells induced by ropivacaine.2.The mechanism of apoptosis induced by ropivacaine may be related to the up-regulation of apoptotic protein Bax,the down-regulation of anti-apoptotic protein Bcl-2,the release of cytochrome C from mitochondria into cytoplasm,the activation of caspase-9 and caspase-3,and the ultimate induction of apoptosis.