花青素对大鼠肝缺血再灌注损伤的作用及其机制
The effect and mechanism of anthocyanin on hepatic ischemia reperfusion injury in rats

摘要：目的 探讨花青素对大鼠肝缺血再灌注损伤的作用及其机制。方法 30只SD 大鼠随机分成假手术组、模型组和花青素组。模型组和花青素组采用血管夹闭肝左中叶血管分支90min再灌注4h构建肝缺血再灌注模型，花青素组在缺血前90min腹腔注射花青素(100mg/kg)，模型组腹腔注射等量生理盐水。再灌注4h后处死大鼠，取肝组织和采集血清。ELISA法检测血清中ALT、AST活性和促炎症因子白介素-6(IL-6)、白介素1β(IL-1β)、肿瘤坏死因子-α(TNF-α)的含量；HE染色观察肝组织病理形态学变化；实时定量PCR法测定IL-6、IL-1β和TNF-α mRNA的水平；硫代巴比妥酸(TBA)法测定丙二醛(MDA)含量；黄嘌呤氧化酶法测定过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GPx)、超氧化物歧化酶(SOD)活性；Western blot法测定肝组织JAK2、STAT3、p-JAK2、p-STAT3和P53蛋白表达。结果 与假手术组相比，模型组ALT、AST活性增高，血清和肝脏中IL-6、IL-1β、TNF-α分泌量和mRNA表达增高，MDA含量增高，p-JAK2、p-STAT3、P53表达增加，肝脏病理改变明显，而CAT、GPx和SOD活性明显降低，JAK2和STAT3表达无明显变化。与模型组相比，花青素组ALT、AST活性降低，血清和肝脏中IL-6、IL-1β、TNF-α分泌量和mRNA表达降低，MDA含量减少，p-JAK2、p-STAT3、P53表达降低，肝脏病理改变减轻，而CAT、GPx和SOD活性明显增加，JAK2和STAT3表达依然无明显变化。结论 花青素可通过抑制氧化应激和炎症减轻肝脏缺血再灌注损伤，其机制可能与抑制JAK2/STAT3/P53信号通路的激活有关。
ABSTRACT: Objective To investigate the effect and mechanism of anthocyanin on hepatic ischemia reperfusion injury in rats. Methods Totally 30 SD rats were randomly divided into sham group, model group and anthocyanin group (n=10 in each). Hepatic ischemia was constructed in model group and anthocyanin group by ligating left middle lobe of liver vascular branches for 90min. Anthocyanin was administered at 90min before ischemia by intraperitoneal injection at the concentration of 100mg/kg for rats in anthocyanin group. Rats in model group and sham group were injected with the same dosage of normal saline at the same time. Serum and liver tissues were collected at 4h after reperfusion by putting the rats to death. The expressions of ALT, AST, IL-6, IL-1β and TNF-α in the serum were examined by ELISA. The pathological changes of liver tissue were evaluated by HE. The mRNA levels of IL-6, IL-1β and TNF-α were measured by RT-PCR. The content of MDA was examined by TBA. The activities of CAT, GPx and SOD were evaluated by xanthine oxidase method and the expression of p-JAK2, p-STAT3, JAK2, STAT3 and P53 were measured by Western blot. Results Compared with those in the sham group, the activities of ALT and AST, the expressions of p-JAK2, p-STAT3, P53, IL-6, IL-1β, TNF-α and the content of MDA as well as the pathological changes of the liver in model group were significantly increased. However, the activities of CAT, GPx and SOD in model group were decreased and the expressions of JAK2 and STAT3 between the two groups did not differ. Compared with those in model group, the activities of ALT and AST, the expressions of p-JAK2, p-STAT3, P53, IL-6, IL-1β, TNF-α, the content of MDA and the pathological changes of the liver in anthocyanin group were significantly decreased. But the activities of CAT, GPx and SOD in