CAJ | 학술논문

目的探讨含饱和氢气生理盐水对小鼠肝脏缺血再灌注(IP)损伤的作用及其机制.方法将C57BL/6小鼠分为3组,每组8只.假手术(SO)组小鼠仅接受开腹及关腹操作;对照组小鼠建立肝脏缺血再灌注损伤模型,并于肝脏缺血同时经尾静脉注射生理盐水5 ml/kg;含饱和氢气生理盐水(HRS)组小鼠肝脏缺血同时经尾静脉注射含饱和氢气生理盐水5 ml/kg.再灌注后6 h处死小鼠,获取静脉血及肝脏组织.检测各组小鼠血清丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)水平;观察肝脏组织形态学变化;分析各组肝脏组织损伤程度;检测各组肝脏组织内丙二醛(MDA)含量;抗F4/80抗原免疫组化技术检测各组肝脏组织内巨噬细胞浸润;髓过氧化物酶(MPO)试剂盒检测各组肝脏组织内中性粒细胞浸润;检测各组肝脏组织内肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、细胞间粘附分子1(ICAM-1)和干扰素诱导蛋白(IP)-10mRNA表达;采用蛋白印迹法检测肝脏组织内核因子(NF)-κB亚单位p65磷酸化(p-p65)水平.结果与对照组相比较,HRS组血清ALT和AST水平较低(P＜0.05),肝脏组织损伤情况明显改善(P＜0.01),肝脏组织内MDA含量明显减少(P＜0.01),巨噬细胞及中性粒细胞浸润明显减弱(P＜0.05,P＜0.01),TNF-α、IL-6、ICAM-1和IP-10 mRNA的表达明显降低,转录因子NF-κB活化程度明显减弱.结论静脉注射含饱和氢气生理盐水能够减轻小鼠肝脏IP损伤,其机制可能与抑制肝脏再灌注后氧化应激反应和炎症反应有关.
목적 탐토함포화경기생리염수대소서간장결혈재관주(IP)손상적작용급기궤제.방법 장C57BL/6소서분위3조,매조8지.가수술(SO)조소서부접수개복급관복조작;대조조소서건립간장결혈재관주손상모형,병우간장결혈동시경미정맥주사생리염수5 ml/kg;함포화경기생리염수(HRS)조소서간장결혈동시경미정맥주사함포화경기생리염수5 ml/kg.재관주후6 h처사소서,획취정맥혈급간장조직.검측각조소서혈청병안산전안매(ALT)화천동안산전안매(AST)수평;관찰간장조직형태학변화;분석각조간장조직손상정도;검측각조간장조직내병이철(MDA)함량;항F4/80항원면역조화기술검측각조간장조직내거서세포침윤;수과양화물매(MPO)시제합검측각조간장조직내중성립세포침윤;검측각조간장조직내종류배사인자α(TNF-α)、백세포개소6(IL-6)、세포간점부분자1(ICAM-1)화간우소유도단백(IP)-10mRNA표체;채용단백인적법검측간장조직내핵인자(NF)-κB아단위p65린산화(p-p65)수평.결과 여대조조상비교,HRS조혈청ALT화AST수평교저(P＜0.05),간장조직손상정황명현개선(P＜0.01),간장조직내MDA함량명현감소(P＜0.01),거서세포급중성립세포침윤명현감약(P＜0.05,P＜0.01),TNF-α、IL-6、ICAM-1화IP-10 mRNA적표체명현강저,전록인자NF-κB활화정도명현감약.결론 정맥주사함포화경기생리염수능구감경소서간장IP손상,기궤제가능여억제간장재관주후양화응격반응화염증반응유관.
Objective To explore the protective effect of hydrogen-rich saline on liver ischemiareperfusion (IR) in mice and the possible mechanisms. Methods Twenty-four C57BL/6 mice were randomly divided into 3 groups: sham-operated group, control group (mice were injected with 5 ml/kg saline by tail vein just before ischemia induction) and hydrogen-rich saline group (mice were injected with 5 ml/kg hydrogen-rich saline). Six hour after reperfusion, the mice were sacrificed and the serum and liver samples undergoing IR injury were collected. The ALT and AST levels in serum were determined and liver histiological damage was also evaluated with Suziki's criteria. Malondialdehyde (MDA) contents in liver samples were measured using specific kits. The infiltration of F4/80 positive macrophage cells was detected by using immunohistochemistry and that of neutrophils with myeloperoxidase (MPO) kits. The mRNA expression of TNF-α, IL-6, ICAM-1 and IP-10 was assayed by using real-time reverse transcription PCR. The activation of transcription factor NF-κB was measured by using Western botting analysis. Results As compared with control group, at the 6th h following reperfusion, mice in hydrogen-rich saline group exhibited lower levels of ALT and AST (P＜0. 05) in serum, milder histological damage (P＜0. 01) and less MDA contents in liver samples (P＜0. 01). The infiltration of macrophages, neutrophils and the mRNA expression of TNF-α, IL-6,ICAM-1 and IP-10 in the liver tissue in hydrogen-rich saline group were reduced as compared with IR group (P＜0. 05 or P＜0. 01). The activation of NF-κB in hydrogen-rich saline group was significantly down-regulated as compared with control group. Conclusion Injection of hydrogen-rich saline via the tail vein can alleviate liver IR injury probably by inhibiting oxidant stress and inflammatory response induced by reperfusion.