Tetramethylpyrazine Affect Serum Fibrinogen To Inhibit B16F10 Experimental Metastasis And Mechanism Study

Background and Purpose:Tumors can be identified as a exotic antigen and killed by the immune system.The mechanism of the immune destruction is complex,involving NK cells,lymphocytes,macrophages and other immune cells related to body’s innate and specific immunecytokines.Although the immune system has a strong destruction effect against tumor,Cell journalrecently has reported that more than 99.7%tumor cells were killed by the body’s immune system just entering the blood,but the rest of the very small number of tumor cells evade immune destruction through a variety of ways to successful hematogenous metastasis.Numerous studies was carried on the mechanism of tumor immune escape,discocering that tumor cells evade immune destruction through modulation of surface antigens,induction of MHC abnormal expression,secretion of immunosuppressive factors,and the formation of the tumor thrombus to cover the epitope and other ways.Tumor emboli formation is one of the important ways that tumor cells escape immune destruction.A great deal of research has been carried on the significant role of tumor embolus in metastasis recent years.The studies have shown the relationship between the formation of the tumor emboli and tumor mediated blood coagulation activate is very close.The tumor cell can activate blood coagulation,activating platelet,promoting the fibrinogen into fiber protein,and fibrinogen can recruit the activated platelet to tumor cell surface.The adhesion between fibrinogen and tumor and activated platelet is the key element of the process of the formation of the tumor emboli,therefore fibrinogen play a crucial role in the formation of the tumor emboli.Our research group has long been dedicated to the anti-metastasis of traditional Chinese medicine for activating blood and dissolving stasis,and activating blood coagulation system is the base step of tumor emboli formation.Traditional Chinese medicine for activating blood circulation and removing blood stasis can affect tumor-mediated blood coagulation,having influence on the adhesion between fibrinogen and tumor cells and activated platelet,affecting the formation of tumor emboli.At the same time a lot of clinical data have confirmed that the hypercoagulability is important for procession of malignant tumor at present.The further explanation will be made to study the relationship between hypercoagulability and tumor malignant procession.TMP is the main active ingredient of the traditional Chinese medicine Chuanxiong,now Ligustrazine as a clinical first-line treatment of cardiovascular disease.Pharmacological effect of TMP is wide,with good inhibition of platelet aggregation,increasing microcirculation,antithrombotic,improving blood rheology,immune function,anti-cancer and so on,but whether its anti-tumor effect may be related with influence on the process and thus affect the tumor,thrombus-mediated immune suppression has not yet been reported.This paper focuses on cancer embolus and Ligustrazine anti-tumor metastasis to view the idea of cancer immunotherapy and provide experimental reference for immunotherapy.Methods:In this study,experimental metastasis was used to study the anti-tumor efficacy of TMP.Clotting time was determined by APTT、PT、TT、FIB.Enzyme-linked immunosorbent assay(Elisa)was carried out to test plasma t-PA、u-PA、PAI、FXIII、fibrinopeptide A(FPA)and immunosuppressive factors IL-6、IL-10 to asses TMP arneliorate hypercoagulability and regulate immunosuppression.Adhesive capacity between fibrinogen and B16F10 was detected by fluorescence microplate reader,including fibrinogen-mediated adhesion of platelet and B16F10 melanoma.The fibrinogen receptor integrin(33 and other proteins expression were determined by western blotting and Immunohistochemistry.Key Results:The results showed TMP reduced the number of lung nodules.Tumor-mediated hypercoagulability could be improved after TMP treatment.TMP can extend the clotting time,inhibiting the formation and stability of fibrin.TMP inhibited adhesion of B16F10 and fibrinogen.Besides,the fibrinogen-mediated adhesion of the activated platelet and B16F10 can be inhibited dramatically.TMP could improve the immune organ index and reduce the release of immunosuppressive factors.Immunohistochemistry of lung tissue suggested TMP could reduce IL-6、TNF-a、NF-kB and E-cadherin expression,and regulation of Integrin β3 and E-cadherin might contribute to the inhibition of adhesion.In conclusions,TMP improved the hypercoagulability,inhibited cancer embolus formation and reduced tumor-induced immune suppression to achieve anti-tumor purposes.Conclusions and Implications:In conclusions,TMP improved the hypercoagulability,inhibited cancer embolus formation and reduced tumor-induced immune suppression to achieve anti-tumor purposes.This property might offer an opportunity to study underlying mechanisms for the tumor immunotherapy.Early in tumor metastasis,the vast majority of tumor cells killed by NK cells,very few tumor cells escape from immune surveillance,and thus be able to transfer in the blood.Therefore,the formative stage of tumor thrombus is a critical period of the tumor process,if this process will be affected which is of great significance for the tumor thearpy.