Study on the Efficacy of Xiongzhi Tongluo Formula in Treating Ischemic Stroke with Stasis Toxin Syndrome in Rats and the Intervention Mechanism of Thrombin Mediated Neurotoxicity

LIU Yue; LIANG Xiao; SHEN Wei; WEI Jingjing; LIU Hongxi; ZHAO Di; CHEN Yunmeng; WANG Liuding; MIAO Lina; ZHANG Yunfan; ZHANG Yunling

Abstract：Objective To investigate the therapeutic effects of Xiongzhi Tongluo formula(XZTLF)on a rat model of ischemic stroke with stasis toxin syndrome(STS),and to explore its molecular mechanism in thrombin-mediated blood-brain barrier(BBB)injury.Methods Ninety male SD rats were randomly divided into six groups(n=15 per group):sham-operated group,model group,argatroban(positive drug)group,and XZTLF low-,medium-,and high-dose groups.Except for the sham-operated group,all other groups were subjected to ischemic stroke with STS using a disease-syndrome combination modeling approach.Neurological function,cerebral infarct area,brain water content,tongue color(RGB values),pulse characteristics,whole blood viscosity,plasma viscosity,and coagulation function(including PT,APTT,TT,and FIB levels)were evaluated.Pathological morphology and ultrastructure of the ischemic penumbra were observed.The expression levels of BBB-related proteins(ZO-1,Occludin)and thrombin pathway markers(Thrombin,PAR-1,MMP-9)were measured.Results Compared with the sham-operated group,the model group showed significantly increased infarct area,brain water content,blood viscosity,and plasma viscosity(P<0.01),along with decreased tongue RGB values and pulse amplitude(P<0.01).Coagulation parameters(PT,APTT,TT)were shortened,and FIB levels were elevated(P<0.05).In the ischemic penumbra,ZO-1 and Occludin expression decreased(P<0.01),while Thrombin,PAR-1,and MMP-9 levels increased(P<0.01).Histopathology revealed neuronal loss,nuclear pyknosis,and disrupted BBB integrity.Compared with the model group,medium-and high-dose XZTLF significantly improved neurological function,reduced infarct size and brain edema(P<0.05),increased tongue RGB values(P<0.05).The high-dose XZTLF group showed significantly increased pulse amplitude(P<0.01).The medium-dose XZTLF significantly prolonged PT,APTT and TT(P<0.05),reduced FIB levels(P<0.05),upregulated ZO-1 and Occludin expression(P<0.05),and downregulated Thrombin,PAR-1 and MMP-9 protein levels(P<0.05)in the ischemic penumbra.Pathological staining revealed increased neuronal numbers and reduced nuclear pyknosis in all treatment groups.Electron microscopy demonstrated marked improvement in BBB ultrastructure across all dosage groups.Conclusion XZTLF significantly ameliorates neurological deficits and STS manifestations in rats with ischemic stroke,potentially by regulating the Thrombin/PAR-1/MMP-9 pathway to suppress thrombin-induced neurotoxicity and alleviate blood-brain barrier damage.