血必净注射液对烫伤大鼠高迁移率族蛋白B1表达和急性肝损伤的影响

Effect of ＂Xuebijing injection＂ on expression of high mobility group box-1 protein and acute liver injury in rats with scald injury

目的了解血必净注射液对烫伤大鼠肝组织高迁移率族蛋白B1（HMGB1）表达及急性肝损伤的影响。方法制作大鼠30％TBSAm度烫伤模型，将78只大鼠按随机数字表法分为假伤组（18只）、烫伤组（30只）和血必净治疗组（30只），分别于伤后8、24、72h处死取材。观察肝组织病理学变化；测定血清丙氨酸转氨酶（ALT）和天冬氨酸转氨酶（AST）水平；RT—PCR法检测肝组织HMGB1 mRNA表达，蛋白质印迹法及免疫组织化学法检测肝组织HMGB1蛋白相对表达量，结果分别以积分吸光度比值、吸光度值表示。结果光学显微镜下可见烫伤组大鼠肝组织有大量炎性细胞浸润，以伤后24h较多；治疗组则明显减少。与假伤组比较，烫伤组大鼠血清ALT、AST水平显著增高（P〈0．05或P〈0．01），肝组织HMGB1基因和蛋白表达于伤后8～72h显著增强（P〈0．05或P〈0．01）。与烫伤组比较，治疗组大鼠血清ALT、AST水平有不同程度下降（P〈0．05或P〈0．01）；伤后24、72h肝组织HMGB1 mRNA表达分别为0．60±0．15、0．55±0．07，蛋白水平分别为163±13、160±16，显著低于烫伤组的0．75±0．12、0．78±0．11和200±13、175±14（P〈0．05或P〈0．01）。结论HMGB1作为晚期炎性因子，参与了严重烫伤大鼠肝组织炎性反应的病理过程。血必净治疗可明显下调HMGB1表达，有助于减轻延迟复苏导致的急性肝损伤。

Objective To investigate the effect of ＂ Xuebijing injection＂ （Xuebijing in brief） on expression of hepatic high mobility group box-1 protein （HMGB1） and acute liver injury in rats with scald injury. Methods Seventy-eight rats were divided into sham scald group （ n = 18） , scald group （ n = 30） , and Xuebijing treatment group （n = 30）. Rats in the latter 2 groups were subjected to 30% full-thickness scald injury followed with delayed resuscitation. These rats were sacrificed at 8th , 24th , and 72nd post-injury hour （PIH） to collect specimens. The hepatic pathological changes were observed. Serum levels of ALT and AST were detected. HMGB1 mRNA level in hepatic tissue was detected by the reverse transcription polymerase chain reaction. Protein relative expression quantity of HMGB1 in hepatic tissue was determined with Western blot and immunohistochemistry. Outcomes were denoted in integral absorbance ratio and absorbanee value respectively. Results Massive infiltration of inflammatory cells in hepatic tissues was observed in scald group under light microscope, especially at 24th PIH, and it was decreased in quantity in Xuebijing treatment group. Compared with those of sham scald group, both mRNA and protein expressions of HMGB1 in hepatic tissue of scald group were significantly enhanced during 8 - 72 PIH （ P 〈 0.05 or P 〈 0. 01 ） , along with markedly increased serum levels of ALT and AST （ P 〈0.05 orP 〈0.01 ）. Compared with those in scald group at 24th and 72na PIH, hepatic HMGB1 mRNA expressions （0.75 ±0. 12 vs. 0.60 ±0. 15 and 0.78±0.11 vs. 0.55 ±0.07, respectively） and protein values （200±13 vs. 163 ±13 and 175 ±14 vs. 160± 16, respectively） in Xuebijing treatment group were markedly down-regulated （ P 〈0.05 or P 〈 0.01 ） , and serum levels of ALT and AST decreased in different degrees （ P 〈 0.05 or P 〈 0. 01）. Conclusions HMGB1, the delay-appearing inflammatory mediator, is involved in the pathogenesis of inflammatory response in hepatic tissue in severely scalded rats. Treatment with Xuebijing can markedly down-regulate hepatic HMGB1 expression and protect liver against acute injury induced by delayed resuscitation.