犀角地黄汤对脓毒症大鼠肝组织氧化磷酸化通路相关基因表达的影响

Effect of Xijiao Dihuang Decoction on the expression of genes related to oxidative phosphorylation pathway in liver tissue of septic rats

目的探讨犀角地黄汤抗脓毒性肝损伤的可能机制。方法将30只Wistar大鼠随机等分为对照组、脓毒症组和脓毒症犀角地黄汤治疗组。采用盲肠结扎穿孔(CLP)法复制脓毒症大鼠模型,对照组按同法手术但不行CLP。各组大鼠术后肌注平衡液5 ml/kg;犀角地黄汤治疗组大鼠于术前2 d开始给予犀角地黄汤(相当于生药0.45 g)灌胃,2次/d,术后继续灌胃1 d。24 h后各组随机取5只大鼠的肝组织提取总RNA组行mRNA测序,应用生物信息学分析犀角地黄汤干预后的大鼠肝组织氧化磷酸化(Oxidative phosphorylation)通路相关基因表达情况。结果对照组大鼠的肝组织结构清晰,脓毒症组大鼠的肝组织有较多的炎症细胞浸润,而脓毒症犀角地黄汤治疗组的大鼠肝组织炎症细胞浸润较脓毒症组明显减轻。脓毒症大鼠24 h肝组织Oxidative phosphorylation通路相关mRNA的表达以抑制为主,即出现表达下调数有21个,表达上调2个。经犀角地黄汤干预后,表达抑制得到明显缓解,即脓毒症组大鼠24 h肝组织Oxidative phosphorylation通路相关mRNA表达下调,而犀角地黄汤组恢复正常表达的有9个:NduFs7,NduFs8、NduFv3、NduFa10、NduFa12、NduFb10、F-type ATP a、Succinate degydrogenase SDHC、V-type ATP a;上调转为正常表达的有2个:NduFa4、V-type ATP c。结论犀角地黄汤对脓毒性肝损伤有保护作用,可能是通过缓解NADH脱氢酶相关调节基因及F型、V型ATP合酶相关基因(F-type ATP a,V-type ATP a)及琥珀酸脱氢酶相关基因(SDHC)的低表达而起到抗脓毒性肝损伤的作用。

Objective To investigate the potential mechanism of Xijiao Dihuang Decoction(XJDHT)against septic liver injury at the gene level.Methods 30 Wistar rats were randomized to control group,sepsis group and sepsis Xijiao Dihuang Decoction treatment group.Sepsis rat model was established through Cecal ligation and perforation.The rats of control group underwent the same surgical procedure,but without CLP.Each group was intramuscularly injected with 5ml/kg of balance solution after surgery.Sepsis XJDHT group began to gavage 0.3 ml of rhinoceros horn Dihuang Decoction(equivalent to 0.45g of crude herbs)two days beginning at the day of operation,twice a day,and then gavage continuously after operation a day.After 24 hours,5 rats were randomly selected from each group to extract total RNA from liver tissue for mRNA sequencing.Bioinformatics was used to analyze the expression of genes related to oxidative phosphorylation pathway in liver of rat after XJDHT treatment.Results In the control group,the rats exhibited a distinct and orderly liver tissue structure.By contrast,the liver tissue of rats in the sepsis group showed significant infiltration of inflammatory cells.Interestingly,in the sepsis group treated with Xijiao Dihuang Decoction,the infiltration of inflammatory cells in the liver tissue was notably reduced compared to the untreated sepsis group.The expression of oxidative phosphorylation pathway related mRNA in 24-hour liver tissue of septic rats was mainly inhibited,that is,21 down-regulated and 2 up-regulated.After the intervention of XJDHT,the inhibition was significantly relieved.Such as:the expression of 9 mRNA related to oxidative phosphorylation pathway in 24-hour liver tissue of septic rats was down-regulated,while in XJDHT group returned to normal expression:NduFs7,NduFs8,NduFv3,NduFa10,NduFa12,NduFb10,F-type ATP a,Succinate degydrogenase SDHC,V-type ATP a.There are 2 genes that have changed from upregulation to normal expression:NduFa4,V-type ATP c.Conclusion XJDHT can resist septic liver injury by alleviating the low expression of NADH dehydrogenase related regulatory genes,F-type and V-type ATP synthase-related genes(F-type ATP a,V-type ATP A)and succinate dehydrogenase related genes(SDHC).