| Objective To understand the role of JAK/STAT signal transduction pathway in hepatic ischemia reperfusion injury(HIRI),observe the perioperative treatment of breviscapine on hepatocyte apoptosis and transcriptional activator-1 during HIRI in SD rats.The effects of transcription facter 1(STAT1),matrix metalloproteinase-2(MMP-2)and matrix metalloproteinase-9(MMP-9)on clinical application of breviscapine for prevention of liver deficiency Blood reperfusion injury provides a more complete theoretical basis.Method 40 Sprague-Dawley male rats were randomly and equally divided into five groups(n=8 per group)as follows: Sham,HIRI1+NS,HIRI1+Bre,HIRI2+NS and HIRI2+Bre groups.Groups 1 and 2 represented ischemia for 20 and 60 min,respectively.The duration time of reperfusion was 6 h.The treatment group received intraperitoneal injection of breviscapine 10 mg·kg-1·d-1 from 1 day to 7 days before operation.The sham operation group and the control group were intraperitoneally injected with normal saline 10 ml· kg-1 ·d-1 from 1 day to 7 days before operation.The classical model of HIRI was established in the control group and the treatment group.The blood and liver samples of different groups were collected to observe,after reperfusion for 6 hours.· Biochemical analyzer to detect the levels of aspartate aminotransferase(AST)and serum alanine aminotransferase(ALT)in each rat;· Observing pathological changes in liver tissue specimens;· Cytokine analysis(TNF ?,IL-6 expression levels);· Real-time quantitative PCR detection of STAT1 m RNA expression in liver tissue;· Western blot analysis of the expression levels of STAT1,MMP-2 and MMP-9proteins in liver tissue.Results:1.The levels of serum ALT and AST were significantly increased in the HIRI+NS group compared with Sham group(P<0.01).The HIRI2+NS group was significantly more increased than the HIRI1+NS group.(P<0.01);The levels of serum ALT and AST were significantly decreased in HIRI1+Bre group compared with HIRI1+NS group(P<0.05),especially HIRI2+Bre group compared with HIRI2+NS group(P<0.01).2.The pathological changes of liver tissue were observed by HE staining.The degree of liver tissue damage was increased in HIRI+NS group compared with Sham group,the difference was statistically significant(P<0.01),and HIRI2+NS group was more damaged than HIRI1+NS group.The degree was more significant and the injury was the heaviest(P<0.01).The degree of liver tissue damage was reduced in HIRI+Bre group compared with HIRI+NS group(P<0.05).3.Cytokine analysis showed that the expression levels of TNF ? and IL-6 were higher in HIRI+NS group than in Sham group(P<0.01),and HIRI2+NS group was more increased than HIRI1+NS group(P<0.01);The expression levels of TNF ? and IL-6in HIRI+Bre group were lower than in HIRI+NS group(P<0.05).4.The expression level of MMP-2 protein in each group was lower,and there was no significant difference between the groups(P>0.05).The expression level of MMP-9protein in HIRI+NS group was higher than in Sham group(P<0.01).The expression level of MMP-9 protein in HIRI2+NS group was significantly higher than that in HIRI1+NS group(P<0.01).The expression level of MMP-9 protein in HIRI+Bre group was lower than that in HIRI+NS group.(P<0.05).5.Compared with Sham group,STAT1 protein and STAT1 m RNA expression levels were significantly increased in HIRI+NS group(P<0.01),and HIRI2+NS group was significantly higher than HIRI1+NS group(P<0.01),The expression level of HIRI+Bre group was lower than the corresponding HIRI+NS group(P<0.05).Conclusion:1.The model of HIRI in rat: The ischemic time has increased,more serious damage to the tissue after 6h of reperfusion;2.Perioperative treatment of breviscapine has the protective effect on HIRI,can reduce hepatocyte damage or apoptosis.The mechanism is likely through inhibiting STAT1 signaling pathway,down-regulating the expression of STAT1 and MMP-9,and reducing the infiltration of inflammatory cytokines such as TNF ? and IL-6.. |
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