摘要
目的观察选择性激动黑皮质素4型受体(MC4R)对脓毒症致急性肝损伤大鼠的影响。方法 64只雄性SD大鼠随机分假手术组(假手术后用PBS处理)、盲肠结扎穿孔(CLP)组(采用CLP建立脓毒症模型)、Ro27-3225处理组(CLP后用Ro27-3225处理)和Ro27-3225假手术对照组(假手术后用Ro27-3225处理),每组16只(10只大鼠观察术后一般状态和72 h生存率,另外6只大鼠于术后24 h采集血液及肝组织标本)。各组分别于术后30 min腹腔注射PBS或Ro27-3225(180μg/kg),术后24 h测定大鼠心率(HR)及平均动脉压(MAP)、大鼠血清中天门冬氨酸氨基转移酶(AST)和丙氨酸氨基转移酶(ALT)水平。动物处死后,HE染色观察肝组织病变,反转录PCR检测肝组织中Toll样受体4(TLR4)、高迁移率族蛋白1(HMGB1)和caspase-3 mRNA水平,免疫组织化学染色检测肝细胞中核因子κB(NF-κB)p65的表达,并分析NF-κB p65核阳性率。结果与假手术组相比,CLP组大鼠72 h生存率和MAP降低,血清中AST和ALT水平明显升高,肝索排列紊乱,肝细胞水肿,有明显炎性细胞浸润,肝组织中TLR4、HMGB1和caspase-3 mRNA水平明显升高,肝细胞中NF-κB p65核阳性率明显升高。与CLP组相比,Ro27-3225处理组大鼠72 h生存率和MAP明显升高,血清中AST和ALT水平明显降低,肝组织结构损伤明显减轻,肝组织中TLR4、HMGB1和caspase-3 mRNA水平降低,肝细胞中NF-κB p65核阳性率明显降低。结论脓毒症能引起肝损伤,选择性激动MC4R可能通过抑制HMGB1/TLR4/NF-κB信号途径,减轻炎症反应,降低脓毒症引起的肝损伤。
Objective To observe the effect of selective activation of melanocortin 4 receptor （MC,4R） on the rats with sepsis-induced acute liver injury. Methods Sixty-four male SD rats were randomly grouped into sham operation group （ PBS treatment after sham operation）, cecal ligation and puncture （CLP） group （sepsis model was established by CLP）, Ro27-3225 treatment group （Ro27-327.5 treatment after CLP）, and Ro27-3225 sham operation control group （Ro27-3225 treatment after sham operation）, 16 rats for each group （ten rats were used to observe general condition and ？2-hour survival after operation. Then, six rats were used to collect blood and liver samples）. These groups were intraperitoneally injected with PBS or Ro27-3225 （ 180 μg/kg） 30 minutes after operation. Heart rate （HR）, mean arterial pressure （MAP）, aspartate transaminase （AST） and alanine transaminase （ALT） were measured 24 hours after operation. After execution of the rats, pathological changes of liver tissues were observed by HE staining. The levels of Toll-like receptor 4 （TLP,4）, high mobility group box 1 （ HMGB1 ） and caspase-3 mRNA in liver tissues were analyzed by reverse transcription PCR. The expression of nuclear factor-KB （NF-KB） p65 in hepatocytes was detected by immunohistochemical staining, which was followed byanalysis of nuclear positive rate of NF-KB p65. Results Compared with the sham operation group, CLP group showed decreased 72-hour survival and MAP, significantly increased levels of AST and ALT, hepatic cords disorder, hepatocyte swelling, and diffuse inflammatory cell infiltration; the levels of TLR4, HMGB1 and caspase-3 mRNA in liver tissues remarkably increased, and the positive rate of NF-KB 1065 in hepatocytes went up as well. However, compared with the CLP group, the Ro27-3225 treatment group was found with obviously increased 72-hour survival and MAP, inhibited levels of AST and ALT, attenuated damage of liver tissues, decreased levels of TLR4, HMGB1 and caspase-3 mRNA in liver tissues, and significantly downregulated positive rate of NF-KB p65 in hepatocytes. Conclusion Sepsis causes liver injury. Selectively activating MC4R can reduce sepsis-induced acute liver injury in rats, which may act via inhibiting HMGB1/TLR4/NF-KB signaling pathway to relieve inflammation response.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2016年第8期1055-1059,1064,共6页
Chinese Journal of Cellular and Molecular Immunology
基金
高校省级自然科学研究重点项目(KJ2014A166)
安徽省高等学校自然科学研究(KJ2015B013by)
安徽省高校科研创新平台团队项目(2016-40)
蚌埠医学院自然科学研究资助重点项目(BYKL1402ZD)
蚌埠医学院自然科学研究项目(Byky1213)
