基于长链非编码RNA高通量测序技术明确糖异平对IGT大鼠的作用靶点及信号通路

Identification of action target and signaling pathway of Tangyiping in IGT rats based on high-throughput sequencing of long-chain non-coding RNA

目的应用长链非编码RNA(lncRNA)高通量测序技术探讨糖异平干预IGT大鼠的lncRNA作用靶点及信号通路。方法SPF级Wistar雄性大鼠,随机分成空白组(n=10),IGT模型组(n=10),糖异平组(n=9),糖异平组给予中药混悬液17.14 g·kg^(-1)·d^(-1)。干预8周后,检测血糖相关指标及胰腺组织形态。lncRNA测序检测三组胰腺组织lncRNA表达,取模型组较空白组差异表达lncRNAs与糖异平组较模型组差异lncRNAs的交集进行靶基因预测并作生物学富集功能分析。结果糖异平可减轻IGT大鼠糖负荷后2h血糖(P<0.05),并可改善胰腺形态。根据筛选标准(FC>2或<0.5,且P<0.05),糖异平逆转IGT的lncRNA作用靶点有69个(NONRATT029906.2、NONRATT005832.2、NON-RATT006368.2等),其中上调的有29个,下调的40个。相关靶基因预测共262个,生物学富集功能分析结果显示,糖异平参与血管生长因子(VEGF)、核因子κB(NF-κB)、脂肪细胞因子、胰岛素分泌、胰岛素抵抗等多条信号通路来治疗IGT。结论糖异平干预IGT过程中具有多成分-多靶点-多通路的特征,可能通过调控69个lncRNA靶点,参与VEGF、NF-κB、脂肪细胞因子、胰岛素分泌、胰岛素抵抗等多条信号途径来发挥抗IGT作用。该研究为进一步探讨糖异平治疗IGT的作用机制提供了依据和方向。

Objective To investigate the action target and signaling pathway of lncRNA interfered by Tangyiping in IGT rats by high throughput sequencing of long chain non-coding RNA(lncRNA).Methods SPF Wistar male rats were randomly divided into blank group(n=10),IGT model group(n=10)and Tangyiping group(n=9).Tangyiping group was given 17.14 g·kg^(-1)·d^(-1)traditional Chinese medicine suspension.After intervention for 8 weeks,blood glucose related indexes and pancreatic tissue morphology were detected.The expression of lncRNA in pancreatic tissues of three groups was detected by lncRNA sequen-cing.The intersection of differentially expressed lncRNAs between model group and blank group and differentially expressed lncR-NAs between Tangyiping group and model group was used to predict the target gene and analyze the biological enrichment func-tion.Results Tangyiping could reduce the blood glucose of IGT rats at 2h after glucose loading(P<0.05)and improve the pan-creatic morphology.According to the screening criteria(FC>2 or<0.5,P<0.05),there were 69 lncRNA targets(NON-RATT029906.2,NONRATT005832.2,NONRATT006368.2,etc.)reversed by Tangyiping in IGT rats,among which 29 targets were up-regulated and 40 targets were down-regulated.A total of 262 related target genes were predicted.The results of biolog-ical enrichment function analysis showed that Tangyiping participated in many signal pathways such as vascular growth factor(VEGF),nuclear factorκB(NF-κB),adipocytokines,insulin secretion and insulin resistance to treat IGT.Conclusion Tan-gyiping has the characteristics such as multi-component,multi-target and multi-pathway in the process of IGT intervention,and may play an anti-IGT role by regulating 69 lncRNA targets and participating in many signal pathways such as VEGF,NF-κB,adipocytokines,insulin secretion and insulin resistance.This study provides a basis and direction for further exploring the mechanism of Tangyiping in the treatment of IGT.