目的 探讨血必净注射液对热射病大鼠肝脏的保护作用并研究相关机制.方法 将24只SPF级SD大鼠随机分为正常对照组、热射病组、血必净组,每组8只.正常对照组大鼠麻醉后置于26 ℃的环境中,不作其他处理,直至实验结束;热射病组大鼠用生理盐水(4 ml/kg,尾静脉注射)预处理后进行热射病建模,建模成功后大鼠转移到22～25 ℃的环境中,恢复6 h;血必净组大鼠用血必净注射液(4 ml/kg,尾静脉注射)预处理后进行热射病建模,建模成功后转移到22～25 ℃的环境中,恢复6 h.各组大鼠达到预设时间后,经腹主动脉采血,酶联免疫吸附分析(enzyme-linked im-munosorbent assay,ELISA)检测血清中炎性因子白细胞介素 1β(interleukin 1 beta,IL-1β)、白细胞介素 18(interleukin 18,IL-18)、白细胞介素 6(interleukin 6,IL-6)、肿瘤坏死因子 a(tumor necrosis factor alpha,TNF-a)和高迁移率族蛋白 B1(high mobility group proteins B1,HMGB1)的水平.定量聚合酶链式反应(quantitative polymerase chain reaction,qPCR)检测肝脏组织中核苷酸寡聚化结构域样受体热蛋白结构域相关蛋白3(nucleotide oligomerization domain like receptor hot protein domain related proteins 3,NLRP3)、半胱氨酸蛋白酶 1(Caspase1)、凋亡相关斑点样蛋白(apoptosis related speckled protein,ASC)、肝脏髓过氧化物酶(myeloperoxidase,MPO)水平.苏木精-伊红染色观察各组大鼠肝脏组织形态的病理变化.检测各组大鼠血清中丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天门冬氨酸氨基转移酶(aspar-tate aminotransferase,AST)水平.结果 热射病组大鼠血清炎性因子TNF-α、IL-6、IL-1β和IL-18水平,大鼠血清及肝脏中HMGB1表达水平,肝脏组织中NLRP3、Caspase1以及ASC的mRNA相对表达水平,肝细胞中MPO水平,血清中ALT与AST的含量,显著高于正常对照组(P均＜0.05);肝小叶结构紊乱,中央静脉淤血且存在血栓,肝细胞水肿且出现脂肪颗粒,肝细胞大面积坏死.而血必净组大鼠血清炎性因子TNF-a、IL-6、IL-1β、IL-18、ALT与AST水平,大鼠血清及肝脏中HMGB1表达水平,肝细胞中MPO水平及组织结构评分,均显著低于热射病组(P均＜0.05);仅见肝细胞水肿,肝小叶结构良好.结论 血必净注射液具有减轻热射病大鼠炎症反应及肝脏损伤的作用,其机制可能与抑制NL-RP3炎性小体活化有关.
Study on Xuebijing Injection Inhibits NLRP3 Inflammasome Activation to Protective Effect on Liver Injury Induced by Heat-stroke in Rats
Objective To investigate the protective effects of Xuebijing injection on the liver of heatstroke rats and explore the underlying mechanisms.Methods A total of 24 SPF SD rats were randomly divided into normal control group,heatstroke group and Xuebijing treatment group,with 8 rats in each.In the normal control group,the anesthe-tized rats were placed in 26 ℃ environment without any treatment until the end of the experiment;the rats in the heat-stroke group were pretreated with normal saline(4ml/kg,tail vein injection)and the heatstroke model was established,after successful modeling,the rats were transferred to the environment of 22-25 ℃ and recovered for 6 h;the rats in the Xuebijing treatment group were pretreated with Xuebijing injection(4 ml/kg,tail vein in-jection)for heatstroke modeling,after successful model-ing,the rats were transferred to the environment of 22-25 ℃ and recovered for 6 h.After reaching the predetermined time,blood samples of rats were collected from the aorta abdominals,and the levels of inflammatory factors interleukin 1 beta(IL-1β),interleukin 18(IL-18),interleukin 6(IL-6),tumor necrosis factor alpha(TNF-α)and high mobility group proteins B1(HMGB1)in serum were measured by enzyme-linked immunosorbent assay(ELISA).The levels of nucleo-tide oligomerization domain like receptor hot protein domain related proteins 3(NLRP3),Caspase1,apoptosis related speckled protein(ASC)and myeloperoxidase(MPO)in liver tissue were determined by quantitative polymerase chain re-action(qPCR).Additionally,hematoxylin-eosin staining was performed to observe the pathological changes in liver tis-sue,and the levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in serum of rats were meas-ured.Results The levels of serum inflammatory factors TNF-α,IL-6,IL-1β and IL-18,the expression levels of HMGB1 in serum and liver,and the relative mRNA expression levels of NLRP3,Caspase1 and ASC in liver tissue,the MPO level in hepatocytes,and the contents of ALT and AST in serum of rats in the heatstroke group were significantly higher than those in normal control group(all P＜0.05);the structure of the hepatic lobules was disordered,central venous conges-tion and thrombus were present,hepatocyte edema and fat granules appeared,and hepatocyte necrosis was widespread.But the levels of inflammatory factors TNF-α,IL-6,IL-1β,IL-18,ALT and AST in serum,the expression levels of HMGB1 in serum and liver,the level of MPO in hepatocytes of Xuebijing treatment group were significantly lower than those in heatstroke group(all P＜0.05),only hepatocytes were edematous and the hepatic lobules were well structured.Conclusion Xuebijing injection has a protective effect on inflammatory response and liver damage in heatstroke rats,and its mechanism may be related to the inhibition of NLRP3 inflammasome activation.
HeatstrokeXuebijing injectionInflammatory responseLiver injuryNucleotide oligomerization do-main like receptor hot protein domain related proteins 3
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