Practical Journal of Organ Transplantation(Electronic Version) ›› 2016, Vol. 4 ›› Issue (1): 18-22.

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Effects of hydrogen-rich saline on autophagy in rats underwent orthotopic liver transplantation

  

  • Online:2016-01-20 Published:2021-05-10

饱和氢气生理盐水对大鼠肝移植缺血 / 再灌注损伤中自噬的调节作用

施东婧1 ,杜洪印 2 ,喻文立 2 ,吴莉 1 ,盛明薇 2 ,翁亦齐 2 ,王永旺 2 ,刘文娜 1,王树森3
  

Abstract:

Objective To investigate the effects of hydrogen-rich saline on autophagy in rats whichundergoing orthotopic liver transplantation (OLT). Methods Healthy male Sprague-Dawley (SD)ratsweighing 220-250 g,were divided into three groups(n = 8 each): sham operated group(Sham group);orthotopicliver transplantation group(OLT group);orthotopic liver transplantation with hydrogen-rich saline treatmentgroup(HS group). Sham Group only underwent simple laparotomy. OLT were performed in OLT and HS group.In group HS,6 ml/kg hydrogen-rich saline was injected through infrahepatic vena cava(IVC)after IVC reperfusion.All rats were sacrificed at 6 h after reperfusion. Hepatic function ,oxidative stress(MDA and SOD)and pathologicalchanges were detected. The expression of LC3- Ⅱ and Beclin-1 were detected by Western Blot. The expressionof caspase-3 and Cyt-c mRNA were detected by RT-PCR. Results Compared with Sham group,the liverfunctions,the levels of oxidative stress,the expression of LC3- Ⅱ,Beclin-1、caspase-3 mRNA and Cyt-c mRNA increased markedly in OLT and HS group(MDA(nmol/mg):1.25±0.13、1.02±0.11 vs. 0.64±0.07,SOD(U/mg): 135.78±10.92、158.81±12.09 vs. 204.13±17.75,caspase-3(2-ΔΔCt):2.02±0.26、1.67±0.17 vs. 1.09±0.18,Cyt-C(2-ΔΔCt):1.96±0.19、1.64±0.18 vs. 0.99±0.17,all P < 0.05). In HS group,the pathological changes,the levels of apoptosis and autophagy were significantly ameliorated compared to group OLT〔MDA(nmol/mg :1.02±0.11 vs. 1.25±0.13,caspase-3 :1.67±0.17 vs. 2.02±0.26,Cyt-C :1.64±0.18 vs. 1.96±0.19,all P < 0.05)〕. Conclusion Hydrogen-rich saline can inhibit autophagy and attenuate ischemia reperfusion injury in rats undergoing OLT.

摘要:

目的 研究饱和氢气生理盐水对大鼠原位肝移植缺血 / 再灌注损伤(IRI)中肝细胞自噬的调节作用。方法 选取体重为 220 ~ 250 g 的健康雄性 SD 大鼠 24 只,采用随机数字表法将其分为 3 组(n 8):假手术组(Sham 组)、原位肝移植组(OLT 组)、饱和氢气生理盐水处理组(HS 组)。Sham 组仅进行开关腹操作。OLT 组和 HS 组建立原位肝移植模型。HS 组在肝下下腔开放即刻经肝下下腔静脉注射 6 ml/kg 含饱和氢气的生理盐水,OLT 组则注射等量生理盐水。再灌注 6 h S 组术毕 6 h 时取材,测定血清中丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)水平,观察肝组织形态学改变,检测氧化应激水平,应用蛋白质免疫印迹试验(Western Blot)检测微管相关蛋白轻链 3(LC3- Ⅱ)和自噬相关因子 Beclin-1 蛋白表达,应用 RT-PCR 检测基因天冬氨酸特异性半胱氨酸蛋白酶 3(caspase-3)和细胞色素 C(Cyt-c)的 mRNA 表达水平。结果 Sham 组相比,OLT 组和 HS 组肝功能和病理损伤明显(P 0.05),MDA 水平升高超氧化物歧化酶(SOD)活性降低显著〔MDA(nmol/mg):1.25±0.13、1.02±0.11 比 0.64±0.07,SOD(U/mg):135.78±10.92、158.81±12.09 比 204.13±17.75,均 P < 0.05〕,凋亡及自噬水平均升高(caspase:2.02±0.26、1.67±0.17 比 1.09±0.18,Cyt-c:1.96±0.19、1.64±0.18 比 0.99±0.17,均 P < 0.05)。HS 组与 OLT 组相比,各项指标均下降〔MDA(nmol/mg):1.02±0.11 比 1.25±0.13,caspase-3(2-ΔΔCt):1.67±0.17 比 2.02±0.26,Cyt-c(2-ΔΔCt):1.64±0.18 比 1.96±0.19,均 P < 0.05〕。结论 饱和氢气生理盐水可通过抑制自噬,减少细胞凋亡,有效缓解肝移植中 IRI。