大黄、赤芍对肝衰竭大鼠血清NO合成和分泌影响研究

Effects of rhubarb and radix paeoniae rubra on the synthesis and secretion of serum NO in rats with liver failure

目的:研究大黄、赤芍对90%肝叶切除后肝衰竭大鼠血清NO合成和分泌的影响。方法:SPF级Wistar大鼠300只随机分为5组即正常组、假手术组、模型组、大黄与赤芍联合组、促肝细胞生长素组,构建大鼠肝衰竭模型,测定各组大鼠血清中NO的含量。100g/L的大黄赤芍注射液孵育成模后的肝细胞2、5、15、30、45、60、90、120min,采用Westernblot的方法检测细胞中Akt、eNOS总蛋白及eNOSSer^(1177)、eNOSThr^(495)磷酸化蛋白表达水平。结果:模型组及大黄赤芍联合组肝细胞血清中NO水平于术后16h、32h出现2次高峰,大黄赤芍联合组16h、32h时血清中NO浓度分别为167.4μmol/L、144.4μmol/L。模型组则分别为105.7μmol/L、90.2μmol/L,两组比较有统计学意义(P<0.05)。100g/L的大黄赤芍注射液作用建模后的肝细胞15min后,p-eNOSS-er^(1177)表达水平到达峰值(3.061±0.349)之后逐渐降低;p-eNOSThr^(495)表达水平30min后降低至(0.357±0.063),与其他时间点相比有统计学意义(P<0.05)。结论:大黄、赤芍可能通过激活AkteNOS-NO信号通路而增强血清中NO的合成和分泌,从而发挥促进肝细胞再生的作用。

Objective：To investigate the effect of rhubarb and radix paeoniae rubra on the synthesis and secretion of serum NO in rats with liver failure after 90% hepatic lobectomy.Methods：SPF 300 wistar rats were randomly divided into 5 groups normal group,sham operation group,model group,rhubarb and red peony combination group,pHGF group,was established by liver failure rat model of liver regeneration,the content of serum detection in NO,the rats were observed after 120 hsurvival rate.Hepatocytes 2,5,15,30,45,60,90,and 120 min were incubated with rhubarb and radix paeoniae rubra injection.The expression levels of Akt,eNOS total protein,eNOSSer1177 and eNOSThr495 phosphorylated protein were detected by westernblot in 0.100 g/L.Results：In the model group and the hepatocyte serum of rhubarb,there were two peaks in the serum of hepatocytes in the liver cells at 16 hand32 hafter surgery.The concentration of NO in the serum at 16 hand 32 hin the combined group of rhubarb and radix paeoniae rubra were 167.4μmol/L and 144.4μmol/L.The concentration of NO in the serum at 16 hand 32 hin the model group were 105.7μmol/L and 90.2μmol/L.The two groups were statistically significant（P〈0.05）.After 15 min in the injection of 0.100 g/L of rhubarb and radix paeoniae rubra,the concentration of p-eNOSSer1177 reached the peak（3.061±0.349）,and then gradually decreased.The concentration of p-eNOSThr495 was significantly reduced to（0.357±0.063）after 30 min,which was compared with other time points（P〈0.05）.Conclusion：Rhubarb and radix paeoniae rubra can enhance the synthesis and secretion of NO in serum by activating Akt-eNOS-NO signaling pathway,thereby promoting the regeneration of hepatocytes.