通腑泄热法中药通过线粒体DNA/Toll样受体9/微小RNA-223通路抑制肝脏缺血再灌注损伤  

Inhibitory effect of"Tongfuxiere"method on hepatic ischemia reperfusion injury via mitochondria,deoxyribonucleic acid/toll-like receptor 9/micro ribonucleic acid-223 pathway

作　　者：苑伟 毛本亮 孙楠楠 严林 黄秀凤 龚家权 郝定盈 裴捷 王百林 Yuan Wei;Mao Benliang;Sun Nannan;Yan Lin;Huang Xiufeng;Gong Jiaquan;Hao Dingying;Pei Jie;Wang Bailin(Department of General Surgery,Guangzhou Red Cross Hospital Affiliated to Jinan University,Guangzhou 510220,China;Clinical Medical College of Guizhou Medical University,Guiyang 550004,China;Dongguan Southeast Central Hospital,Dongguan 523710,China)

机构地区：[1]暨南大学附属广州市红十字会医院普通外科,广州510220 [2]贵州医科大学临床医学院,贵阳550004 [3]东莞市东南部中心医院普通外科,523710

出　　处：《中华实验外科杂志》2022年第3期459-463,共5页Chinese Journal of Experimental Surgery

基　　金：2020年东莞市科学技术局社会科技发展重点项目(202050715032087)。

摘　　要：目的探讨通腑泄热法中药通过线粒体DNA(mtDNA)/Toll样受体9(TLR9)/微小RNA(miR)-223通路抑制肝脏缺血再灌注损伤(HIRI)的作用。方法随机数字法将80只健康无特定病原体(SPF)级SD大鼠分成通路正常组(NP组)、通路激活组(AP组)、通路正常中药组(NPT组)、通路激活中药组(APT组)4组,"通路激活"指在大鼠HIRI建模麻醉后采用TLR9激活剂50μg/只腹腔注射激活mtDNA/TLR9/miR-223通路,"通路正常"则腹腔注射对应剂量生理盐水;"中药"指手术处理前3 d以通腑泄热法中药按照5 ml/只/次剂量,每日2次给大鼠灌胃。每组20只,均建立肝脏HIRI模型,以分组要求施加不同干预后分别于再灌注后24、48 h两个时间点采集血清及肝组织标本,并进行相关生化指标、病理、蛋白质印迹法(Western blot)及聚合酶链反应(PCR)检测。采用多因素方差分析。结果"通路激活"及"中药干预"均是术后血清谷丙转氨酶(ALT)、谷草转氨酶(AST)和乳酸脱氢酶(LDH)变化的影响因素。通路激活可促进术后血清中ALT、AST、LDH升高,AP组大鼠术后24 h血清ALT、AST、LDH高于NP组[ALT:(962.34±42.35)U/L比(747.33±40.04)U/L,F=11.446,P<0.05;AST:(1421.58±93.25)U/L比(1041.87±48.60)U/L,F=5.893,P<0.05;LDH:(2312.99±95.77)U/L比(1974.23±75.80)U/L,F=5.019,P<0.05];而中药干预可抑制术后血清中ALT、AST、LDH升高,NPT组大鼠术后24 h血清ALT、AST、LDH低于NP组[ALT:(561.31±40.49)U/L比(747.33±40.04)U/L,F=688.807,P<0.05;AST:(792.38±44.03)U/L比(1041.87±48.60)U/L,F=838.255,P<0.05;LDH:(1702.75±62.32)U/L比(1974.23±75.80)U/L,F=514.608,P<0.05];通路激活及中药干预的交互作用呈现出抑制作用,APT组大鼠术后24 h血清ALT、AST、LDH低于NP组[ALT:(437.88±47.88)U/L比(747.33±40.04)U/L,F=156.296,P<0.05;AST:(510.02±55.54)U/L比(1041.87±48.60)U/L,F=272.576,P<0.05;LDH:(1473.69±71.94)U/L比(1974.23±75.80)U/L,F=134.480,P<0.05];且术后48 h,NP组、AP组、NPT组、APT组4组大鼠血清ALT、AST、LDH均较术后24 Objective To investigate whether liver ischemia reperfusion injury(HIRI)can be inhibited by Chinese traditional medicine"Tongfuxiere"method through the mitochondria,deoxyribonucleic acid(mtDNA)/toll-like receptor 9(TLR9)/micro ribonucleic acid(miR)-223 pathways.Methods A total of 80 healthy specific pathogen free(SPF)SD rats were randomly divided into 4 groups:the pathway-normal group(NP group),the pathway-activated group(AP group),the pathway-normal TCM group(NPT group)and the pathway-activated TCM group(APT group),with 20 rats each group.The HIRI experiment was performed on all rats.Serum and liver tissue samples were then collected at two time points:24 h and 48 h after reperfusion,after applying different interventions according to group requirements.These specimens were used for biochemical,pathological,Western blotting,and PCR examinations.Results were analyzed by SPSS 25.0 statistical software.Measurement data meeting normal distribution were expressed as mean±standard deviation(SD),using multi-way ANOVA.P<0.05 was used as a statistically significant difference.Results"Pathway activation"and"TCM intervention"were all the factors influencing the postoperative changes in serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),and lactate dehydrogenase(LDH).Pathway activation could promote the elevation of ALT,AST,and LDH in the postoperative serum.Serum ALT,AST and LDH were higher at 24 h after surgery in the AP group than in the NP group[ALT:(962.34±42.35)vs.(747.33±40.04)U/L,F=11.446,P<0.05;AST:(1421.58±93.25)vs.(1041.87±48.60)U/L,F=5.893,P<0.05;LDH:(2312.99±95.77)vs.(1974.23±75.80)U/L,F=5.019,P<0.05].TCM intervention could inhibit the elevation of serum ALT,AST,and LDH postoperatively.Serum ALT,AST and LDH at 24 h after surgery in the NPT group were lower than in the NP group[ALT:(561.31±40.49)vs.(747.33±40.04)U/L,F=688.807,P<0.05;AST:(792.38±44.03)vs.(1041.87±48.60)U/L,F=838.255,P<0.05;LDH:(1702.75±62.32)vs.(1974.23±75.80)U/L,F=514.608,P<0.05].The interaction effects of pathway a

关 键 词：肝脏缺血 再灌注损伤 通腑泄热法 

分 类 号：R285.5[医药卫生—中药学]