Study On The Mechanism Of Jiedu Huoxue Formula On Smooth Muscle Cell Proliferati On In TLR4/NF-κB Signaling Pathway Intervening In Inflammatory Response

Objective:In recent years,with the development and application of interventional diagnosis and treatment technology,percutaneous coronary intervention(PCI),as one of the most representative interventional treatment methods,can effectively improve the clinical symptoms of patients with acute coronary syndrome.and prognosis.As an exogenous substance,stents lead to in-stent restenosis(ISR)by promoting the proliferation,migration,immune response of vascular smooth muscle cells and the formation of neointima after implantation.Inflammatory mechanism is one of the main core mechanisms leading to the formation of in-stent restenosis.Toll-like receptor 4/nuclear factor-κB(TLR4/NF-κB)signal transduction pathway may be involved in the process of intimal hyperplasia and restenosis.plays an indispensable role.Combined with the research progress of modern medicine in recent years,we found that the etiology and pathogenesis of "toxicity and blood stasis" in TCM is closely related to the "inflammation theory",one of the core mechanisms of vascular restenosis.In this experiment,from the cellular level,by establishing an in-stent restenosis inflammatory cell model in vitro,to simulate the effect of Jiedu Huoxue Formula on the inhibition of inflammatory response and vascular smooth muscle cell proliferation in patients after PCI in terms of mediating TLR4/NF-κB signal transduction pathway It can provide a theoretical basis for the smooth development of related clinical treatment.Methods:In this experiment,TNF-α(tumor necrosis factor-α)was used to induce inflammatory response in A7r5 cells(rat thoracic aortic smooth muscle cells)to simulate restenosis inflammatory cells in stents in vitro.Aspirin-containing serum,TLR4 inhibitor and NF-The κB inhibitor was used as the control group,and the mechanism of Jiedu Huoxue Formula intervening in inflammation through the TLR4/NF-κB pathway and inhibiting the proliferation of smooth muscle cells was preliminarily explored at the cellular level.In this experiment,the drug-containing serum was prepared first,and then the toxicity and optimal inhibitory concentration of the drug-containing serum of Jiedu Huoxue Formula were determined.Finally,the inhibitory effect of the drug-containing serum of Jiedu Huoxue Formula on the proliferation activity of A7r5 cells and the levels of inflammatory factors TNF-αand IL-1β was observed.The protein expressions of TLR4,NF-κB and My D88 were determined by western blot.Results:1.Compared with the blank group,the cell proliferation activity of the TNF-α-induced group was significantly increased(P<0.05).Compared with the TNF-α-induced group,the aspirin serum group and the Jiedu Huoxue Formula serum group could reduce the proliferation activity of A7r5 cells in both groups(P<0.05).2.Compared with the blank group,the contents of inflammatory factors TNF-α and IL-1β were significantly increased in the TNF-αinduction group,Jiedu Huoxuefang serum group,aspirin serum group,TLR4 inhibitor group and NF-κB inhibitor group(P<0.05).Compared with the TNF-α-induced group,the levels of TNF-α and IL-1β in the Jiedu Huoxue Formula serum group,aspirin serum group,TLR4 inhibitor group and NF-κB inhibitor group were significantly decreased(P<0.05).Compared with Jiedu Huoxue Formula serum group,the levels of TNF-αand IL-1β in aspirin serum group were significantly lower(P<0.05).3.Compared with the blank group,the expressions of TLR4,NF-κB,and My D88 proteins in the TNF-α-induced group were up-regulated,and both Jiedu Huoxue Formula-containing serum and aspirin-containing serum could down-regulate the protein expressions of TLR4,NF-κB,and My D88.The results are as follows:1.TNF-α can induce A7r5 cells to simulate restenosis inflammatory cells in vitro,which provides a theoretical basis for the study of restenosis after PCI.2.TNF-α can enhance the proliferation activity of A7r5 cells,and Jiedu Huoxue Formula can inhibit the proliferation activity of A7r5 cells.3.TNF-α can induce A7r5 cells to produce inflammatory response,Jiedu Huoxue Formula has anti-inflammatory effect,and the effect is slightly weaker than that of aspirin tablets.4.Jiedu Huoxue Formula can down-regulate the expression of TLR4,NF-κB and My D88 proteins,and inhibit the proliferation and migration of smooth muscle cells.