柴胡皂苷d通过抑制炎性反应减轻大鼠肝脏缺血再灌注损伤  

Saikosaponin d attenuates hepatic ischemia-reperfusion injury by inhibiting inflammatory response in rats

作　　者：朱山飞 张建松 郝定盈 苑伟 吴帆[1] 王百林 Zhu Shanfei;Zhang Jiansong;Hao Dingying;Yuan Wei;Wu Fan;Wang Bailin(Department of General Surgery,Guangzhou Red Cross Hospital of Jinan University,Guangzhou 510220,China;Department of Gastrointestinal Surgery,Guizhou Hospital of The First Affiliated Hospital of Sun Yat-sen University,Guiyang 550000,China;College of Clinical Medicine,Guizhou Medical University,Guiyang 550004,China)

机构地区：[1]暨南大学附属广州红十字会医院普通外科,广州510220 [2]中山大学附属第一医院贵州医院胃肠外科,贵阳550000 [3]贵州医科大学临床医学院,贵阳550004

出　　处：《中华实验外科杂志》2024年第9期2006-2010,共5页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81974442);广东省自然科学基金(2020A1515010799)。

摘　　要：目的观察柴胡皂苷d(SSd)预处理对大鼠肝缺血再灌注损伤(HIRI)炎性反应和Toll样因子受体4(TLR4)/核因子-κB(NF-κB)传导通路的影响.方法随机数字表法将24只雄性SD大鼠分为假手术组(Sham组)、缺血再灌注组(I/R组)和SSd预处理组(SSd组),每组8只.SSd组在术前连续5 d腹腔注射2mg/kg SSd,Sham组和I/R组腹腔注射等量浓度的二甲基亚砜(DMSO).随后,I/R组和SSd组采用Pringle法阻断第一肝门1 h,Sham组只行开腹、关腹手术和肝门解剖等操作.术后6 h,采集大鼠的血清及肝标本.生化检测血清的谷丙转氨酶(ALT)、谷草转氨酶(AST)值;酶联免疫吸附试验(ELISA)法检测血清肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β水平;苏木精-伊红(HE)染色观察肝组织细胞损伤和坏死情况;实时定量反转录聚合酶链反应(RT-qPCR)检测肝脏TLR4、TNF-α及IL-1β的相对表达水平;蛋白质印迹法(Western blot)检测TLR4、MyD88和NF-κB在3组肝组织的表达差异.结果血清学指标:I/R组ALT、AST、TNF-α及IL-1β高于Sham组[(1526.16±147.42)U/L比(71.32±10.21)U/L、(2705.17±127.81)U/L比(189.78±20.04)U/L、(48.60±23.03)pg/ml比(6.67±1.72)pg/ml、(119.18±40.35)pg/ml比(5.55±3.61)pg/ml,t=27.85、54.99、5.13、7.93,P<0.01];而SSd组上述指标低于I/R组[(402.79±67.54)U/L比(1526.16±147.42)U/L、(658.89±88.35)U/L比(2705.17±127.81)U/L、(11.38±5.08)pg/ml比(48.60±23.03)pg/ml、(24.82±5.98)pg/ml比(119.18±40.35)pg/ml,t=19.59、37.25、4.63、6.54,P<0.01],差异有统计学意义.HE染色观察肝组织病理改变及评分:给予SSd干预后,SSd组肝脏组织损伤轻于I/R组,Suzuki评分差异有统计学意义(5.40±1.67比8.00±0.71,t=3.20,P<0.01).进一步通过RT-qPCR结果发现,I/R组TLR4、TNF-α及IL-1β的mRNA相对表达量明显高于Sham组(2.45±0.32比1.00±0.00、1.96±0.17比1.00±0.00、1.63±0.19比1.00±0.00,t=4.63、7.93、5.74,P<0.01),而SSd组TLR4、TNF-α及IL-1β的mRNA相对表达量低于I/R组(1.92±0.20比2.45±0.32、1.55Objective 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)-1βlevels.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-1βin 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-1βwere higher in the I/R group than in the Sham group[(1526.16±147.42)U/L vs.(71.32±10.21)U/L,(2705.17±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 th

关 键 词：柴胡皂苷 肝缺血再灌注损伤 炎性反应 Toll样因子受体4 核因子-ΚB 

分 类 号：R285.5[医药卫生—中药学]