香叶木素预处理通过抗炎和抗氧化发挥对小鼠肝缺血/再灌注的保护作用

The protective effect of diosmetin on liver ischemia/reperfusion in mice though anti-inflammation and antioxidation

目的研究香叶木素(diosmetin,Dio)在肝脏缺血/再灌注(ischemia/reperfusion,I/R)中的作用及其可能的机制。方法32只小鼠随机分为假手术组、I/R组、低剂量Dio组和高剂量Dio组,每组8只。I/R组行肝I/R术,假手术组行假手术。低和高剂量Dio组行肝I/R术前30 min分别腹腔注射10 mg·kg^(-1)和40 mg·kg^(-1) Dio。再灌注后24 h,采集血液和肝脏样本。ELISA检测血清中白介素(IL)-1β、IL-6、乳酸脱氢酶(LDH)和天冬氨酸氨基转移酶(AST)的水平。测定肝匀浆中丙二醛(MDA)、活性氧(ROS)、还原性谷胱甘肽(GSH)和总胆红素(TBIL)的水平。HE染色观察肝损伤情况。免疫组化染色观察肝组织中IL-1β的表达。Western blot检测肝组织中cleaved-caspase-3、p-NF-κB p65和p-p38蛋白表达。结果与假手术组比较,I/R组小鼠肝损伤程度明显增加,血清中IL-1β、IL-6、LDH和AST水平、肝组织中MDA和ROS水平以及cleaved-caspase-3、p-NF-κB p65和p-p38蛋白水平均明显升高,而GSH和TBIL的水平明显下降,差异均具有统计学意义。与I/R组比较,低和高剂量Dio组中上述指标均得到明显改善(P<0.05),其中高剂量组的效果优于低剂量组。结论Dio预处理可减轻肝I/R损伤,这一作用可能与其减弱肝I/R诱导的氧化应激以及炎症相关通路有关。

Aim To study the role of diosmetin(Dio)in liver ischemia/reperfusion and its possible mechanism.Methods Thirty-two mice were randomly divided into sham group,I/R group,low-dose Dio group and high-dose Dio group,with 8 mice in each group.The I/R group received liver I/R surgery,and the sham group performed sham surgery.The low-and high-dose Dio groups received intraperitoneal injection of 10 mg·kg^(-1) and 40 mg·kg^(-1) Dio at 30 min before liver I/R,respectively.Blood and liver samples were collected at 24 h after reperfusion.ELISA was used to detect the levels of interleukin(IL)-1β,IL-6,lactic dehydrogenase(LDH)and aspartate transaminase(AST)in serum.The levels of malondialdehyde(MDA),reactive oxygen species(ROS),reduced glutathione(GSH)and total bilirubin(TBIL)in liver homogenate were measured.HE staining was used to observe liver injury.Immunohistochemical staining was used to observe the expression of IL-1β in liver tissues.Western blot was used to detect the expression of cleaved-caspase-3,p-NF-κB p65 and p-p38 proteins in liver tissues.Results Compared with sham group,the degree of liver injury in I/R group significantly increased,and the levels of IL-1β,IL-6,LDH and AST in serum,MDA,ROS and cleaved-caspase-3,p-NF-κB p65 and p-p38 proteins in liver tissues markedly increased;however,the levels of GSH and TBIL significantly decreased,and the differences in the above indexes were statistically significant.Compared with I/R group,the above indexes in low-and high-dose Dio groups were significantly improved,and the effect of high-dose group was better than that of low-dose group.Conclusions Dio pretreatment can reduce liver I/R injury,which may be related to the reduction of oxidative stress and inflammation-related pathways induced by liver I/R.