| Objectives 1.To investigate the effects of TF and PARs in rat monocytes of peripheral blood stimulated with LPS in vitro.2.To observe effects of Xuebijing injection on expression of TF,IL-6,TNF-αand PAR-1,PAR-2 in rat monocytes stimulated with LPS in vitro.3.To investigate the effects of Xuebijing injection on PAR-1,PAR-2 of monocyte and plasma TF,IL-6,TNF-αin septic rats.4.To observe the effects of Xuebijing injection on coagulation parameters and PLT in septic rats.Methods 1.The monocytes were isolated from the peripheral blood of male Wistar rats,were suspended in cell culture plates,then were stimulated by 1μg/ml LPS. Compared with APC,exposuring to the 10mg/ml Xuebijing injection for different duration(12,24,48 and 72h) the cell supernatants were harvested to determine the levels of TF,IL-6 and TNF-αby ELISA,the monocytes were denatured in cell culture plates to determine PAR-1,PAR-2 by FCM.2.The monocytes were stimulated by 1μg/ml LPS then exposuring to different concentration Xuebijing injection(1,10 and 100mg/ml) for 24h,the cell supernatants were harvested to determine the levels of TF,IL-6,TNF-αby ELISA,the monocytes were denatured in cell culture plates to determine PAR-1,PAR-2 by FCM.3.One hundred and four Wistar rats were randomly divided into four groups:normal group,sham-operation group,CLP group, and Xuebijing-treated group.Blood samples of each group were obtained from abdominal aorta after 12,24,48 and 72h of operation.PAR-1,PAR-2 of monocyte was assessed by FCM,the levels of plasma TF,IL-6 and TNF-αwas determined by ELISA, the expression of TF and IL-6 mRNA of the peripheral blood mononuclear cells was determined by RT-PCR and the PT,APTT,FIB and PLT were assessed.Results 1.In vitro,compared with normal group,PAR-1 up-regulated in LPS group (P<0.01),PAR-2 up-regulated after each time(P<0.05 or P<0.01),TF,IL-6 and TNF-αin the supernatant significantly increased after each time(P<0.01).Compared with LPS group,PAR-1 down-regulated in LPS+APC group after 24,48 and 72h(P<0.01), PAR-2 up-regulated after 24 and 48h(P<0.01),TF,IL-6 and TNF-αdecreased after each time(P<0.05 or P<0.01).Compared with LPS group,PAR-1 down-regulated in LPS+ Xuebijing injection group after 24,48 and 72h(P<0.01),PAR-2 up-regulated after each time(P<0.01),TF,IL-6 and TNF-αdecreased after each time(P<0.05 or P<0.01).2.In vitro,Compared with LPS group,PAR-1 down-regulated(P<0.01) and TF,IL-6 and TNF-αdecreased(P＜0.05 or P＜0.01) treated with 1,10 and 100mg/ml Xuebijing injection.It's obvious in 10mg/ml group.PAR-2 up-regulated(P＜0.01) in 10mg/ml group.3.Compared with normal group and sham-operation group,PAR-1 up-regulated in CLP group after 48 and 72h(P＜0.05 or P＜0.01),PAR-2 up-regulated after each time(P＜0.05 or P＜0.01),TF increased after 24,48 and 72h(P＜0.05 or P＜0.01),IL-6 and TNF-αincreased after each time(P＜0.05 or P＜0.01),TF and IL-6 mRNA increased after 12,24 and 48h(P＜0.05 or P＜0.01),PT shortened after 24,48 and 72h(P＜0.05 or P＜0.01),APTT extended after 12 and 48h(P＜0.01),FIB decreased after 12,24 and 48h(P＜0.05 or P＜0.01),PLT decreased after 12 and 24h(P＜0.05 or P＜0.01).Compared with CLP group,PAR-1 down-regulated in Xuebijing injection group after 12 and 24h(P＜0.05 or P＜0.01),PAR-2 up-regulated after each time(P＜0.05 or P＜0.01),TF decreased after 24,48 and 72h(P＜0.05 or P＜0.01),IL-6 decreased after 24 and 48h(P＜0.01),TNF-αdecreased after each time(P＜0.05 or P＜0.01),TF mRNA decreased after 12,24 and 48h(P＜0.05 or P＜0.01),IL-6 mRNA decreased after each time(P＜0.05 or P＜0.01),PT extended after 24 and 48h(P＜0.05 or P＜0.01),APTT shortened after 48h(P＜0.05),FIB increased after 12,24 and 48h(P＜0.01),PLT increased after 24 and 48h(P＜0.05 or P＜0.01).Conclusions 1.Xuebijing injection and APC could decrease LPS-stimulated PAR-1 expression on monocytes and TF,IL-6 as well as TNF-αrelease in supernatants.It's the best after 24 and 48h treated by 10mg/ml Xuebijing injection.Xuebijing injection was better than APC in the experiment 2.Xuebijing injection would obviously decreas the plasmic IL-6 and TNF-α,ameliorate the coagulation dysfunction in sepsis by reducing monocytic TF and down-regulating monocytic PAR-1.
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