基于RNA-Seq技术分析小半夏汤抑制1-PBG诱导的大鼠异食癖行为的转录组学特征

Transcriptionomic Characteristics of Xiaobanxiatang(XBXT)against 1-PBG Induced Emesis in Rat Pica Model by RNA-Seq Technique

目的:采用选择性5-羟色胺3(5-HT3)受体激动剂1-phenylbiguanide(1-PBG)建立大鼠异食癖模型,通过转录组测序(RNA-Seq)技术分析小半夏汤止吐作用的转录组学特征,以进一步探讨其通过阻断5-HT3受体防治化疗性恶心呕吐(chemotherapy induced nausea and vomiting,CINV)的作用机制。方法:腹腔注射1-PBG(25 mg/kg,10 ml/kg)建立大鼠异食癖模型,给予小半夏汤灌胃(3.2 g/kg,每天分两次给予),连续灌胃3 d,每天记录大鼠高岭土摄入量。采用RNA-Seq技术检测回肠组织中差异基因表达情况。结果:与正常对照组比较,模型对照组大鼠摄食高岭土量显著增加(P<0.01),小半夏汤3.2 g/kg组能够显著抑制模型对照组大鼠摄食高岭土(P<0.01)。转录组学测序分析结果显示,与正常对照组比较,模型对照组大鼠回肠组织中有759个基因明显上调(P<0.05),423个基因显著下调(P<0.05)。小半夏汤逆转了154个上调基因(P<0.05),这些基因主要参与白细胞跨内膜迁移、Th1和Th2细胞分化等炎症相关信号通路;逆转了92个下调基因(P<0.05),这些基因主要参与维生素消化与吸收、脂肪消化与吸收、PPAR信号通路等物质代谢相关信号通路。结论:小半夏汤可以通过阻断5-HT3受体从而发挥止吐作用。抑制炎症反应相关信号通路的激活、改善机体对营养物质的消化与吸收功能,可能是小半夏汤防治CINV的潜在作用机制。

Objective:To establish the rat pica model by a selective 5-HT3 receptor 1-phenylbiguanide(1-PBG),and explore the transcriptional characteristics of Xiaobanxiatang(XBXT)against emesis by high-throughput RNA-sequencing analysis,in order to study its mechanism of treating and preventing chemotherapy induced nausea and vomiting(CINV)by blocking 5-HT3 receptor.Methods:1-PBG(25 mg/kg,10 ml/kg,i.p.)was used to establish the pica model in rats.XBXT(1.6 g/kg,10 ml/kg,i.g.,b.i.d.)was administered for 3 days,the daily intake of kaoline was observed.The differentially expressed genes in ileum of rats induced by 1-PBG were detected by high-throughput RNA-sequencing.Results:The results showed,compared with the control group,the daily intake of kaoline was significantly increased in the model group(P<0.01).The sequence technique results showed,compared with the control group,759 genes were significantly up regulated(P<0.05)and 423 genes were significantly down regulated in the model group(P<0.05).154 up regulated genes were reversed by XBXT,which were associated with leukocyte transendothelial migration and inflammatory signal pathway like Th1 and Th2 cell differentiation.At the same time,92 down regulated genes were reversed by XBXT,which were associated with vitamin digestion and absorption,fat digestion and absorption and PPAR signaling pathways.Conclusion:XBXT has a certain antiemetic effect by inhibiting 5-HT 3 receptor.Its potential mechanism on CINV may be related to inhibiting the activation of inflammatory signal pathway,and improving the digestion and absorption of vitamin.