Saikosaponin d attenuates hepatic ischemia-reperfusion injury by inhibiting inflammatory response in rats
Objective To investigate the effects of Saikosaponin d(SSd)pretreatment on the inflammatory response and Toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)signaling pathway in rats with hepatic ischemia-reperfusion injury(HIRI).Methods Totally,24 male SD rats were randomly divided into three groups using a random number table:Sham group,ischemia-reperfusion group(I/R group),and SSd pretreatment group(SSd group),with 8 rats in each group.The SSd group received intraperitoneal injections of 2 mg/kg SSd for 5 consecutive days before surgery,and the Sham and I/R groups received equal concentrations of dimethyl sulfoxide(DMSO).Subsequently,the I/R and SSd groups underwent Pringle's maneuver to occlude the hepatic portal vein for 1 h,and the Sham group only underwent laparotomy and hepatic portal anatomy without occlusion.At 6 h post-surgery,serum and liver samples were collected.Biochemical analysis was performed to measure serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels.The enzyme linked immunosorbent assay(ELISA)was used to detect serum tumor necrosis factor-a(TNF-α)and interleukin(IL)-1β levels.The hematoxy-lin and eosin(HE)staining was utilized to observe liver tissue damage and necrosis.The real-time quanti-tative reverse transcriptase-polymerase chain reaction(RT-qPCR)was conducted to measure the relative expression levels of TLR4,TNF-α,and IL-1β in the liver.Western blotting was used to assess the expres-sion differences of TLR4,MyD88,and NF-κB proteins among the three groups.SPSS and GraphPad Prism were used to perform variance analysis on the liver indices of the three groups of rats,and pairwise compar-isons were performed.Results ALT,AST,TNF-α,and IL-1β were higher in the I/R group than in the Sham group[(1 526.16±147.42)U/L vs.(71.32±10.21)U/L,(2 705.17±127.81)U/L vs.(189.78±20.04)U/L,(48.60±23.03)pg/ml vs.(6.67±1.72)pg/ml,(119.18±40.35)pg/ml vs.(5.55±3.61)pg/ml,t=27.85,54.99,5.13,7.93,P＜0.01],while the levels of these markers were lower in the SSd group than in the I/R group[(402.79±67.54)U/L vs.(1 526.16±147.42)U/L,(658.89±88.35)U/L vs.(2 705.17±127.81)U/L,(11.38±5.08)pg/ml vs.(48.60±23.03)pg/ml,(24.82±5.98)pg/ml vs.(119.18±40.35)pg/ml,t=19.59,37.25,4.63,6.54,P＜0.01],with sta-tistically significant differences.After SSd intervention,the liver tissue injury in the SSd group was less se-vere than that in the I/R group,and the difference in Suzuki score was statistically significant(5.40±1.67 vs.8.00±0.71,t=3.20,P＜0.01).Furthermore,the RT-qPCR results showed that the relative mRNA expression levels of TLR4,TNF-α,and IL-1β in the I/R group were significantly higher than those in the Sham group(2.45±0.32 vs.1.00±0.00,1.96±0.17 vs.1.00±0.00,1.63±0.19 vs.1.00±0.00,t=4.63,7.93,5.74,P＜0.01),while the relative mRNA expression levels of TLR4,TNF-α,and IL-1β in the SSd group were lower than those in the I/R group(1.92±0.20 vs.2.45±0.32,1.55±0.05 vs.1.96±0.17,1.24±0.07 vs.1.63±0.19,t=3.45,3.97,3.36,P＜0.05),with statistically significant differences.Finally,the Western blotting results suggested that the expression of TLR4,MyD88,and NF-κB proteins in the I/R group was higher than that in the Sham group(t=27.95,8.67,4.26,P＜0.01),while the expression of TLR4,MyD88,and NF-κB proteins in the SSd group was lower than that in the SSd group(t=3.69,9.43,2.86,P＜0.05),with statistically significant differences.Conclusion SSd pretreatment significantly reduces the inflammatory response in the rat HIRI,potentially through the downregulation of TLR4/NF-κB signaling pathway proteins.
NETL
NSTL
万方数据
