通络地龟汤对1型糖尿病小鼠肾脏lncRNA、mRNA表达谱的影响

Effect of Tongluo Digui Decoction on Expression Profile of lncRNA and mRNA in Kidney of Type 1 Diabetic Disease Mice

目的:通过对糖尿病肾病小鼠肾组织长链非编码RNA(long noncoding RNA,lncRNA)和信使RNA(mRNA)表达谱的检测,探讨通络地龟汤(TLDGD)治疗糖尿病肾病的作用机制。方法:C57BL/6N小鼠随机分为正常组、DKD模型组、DKD+TLDGD组,TLDGD灌胃8周后检测血肌酐(serum creatinine,Scr)、尿素氮(blood urea nitrogen,BUN)、24 h尿蛋白(24 h protein,24 hPro)、空腹血糖(fasting blood glucose,FBG);HE染色、Masson染色、PAS染色观察各组小鼠肾脏病理改变;应用基因芯片技术,检测与TLDGD治疗糖尿病肾病作用密切相关的差异lncRNA和mRNA,进行聚类分析。运用生物信息学方法预测差异lncRNA调控的靶基因,并对其进行功能和通路分析。结果:与正常组相比,DKD模型组Scr、BUN、24 hPro、FBG均升高,差异均有统计学意义(P<0.01),与DKD模型组相比,DKD+TLDGD组上述指标均降低,差异均有统计学意义(P<0.05),DKD+TLDGD组肾脏病理损伤明显改善;TLDGD调控了84个lncRNA差异表达,其中上调42个,下调42个,差异均有统计学意义(P<0.05)。差异表达的mRNA共189个,其中上调132个、下调57个,差异均有统计学意义(P<0.05);使用GO分析方法,发现下调或上调的靶基因主要参与了炎症中的细胞迁移、炎症反应、细胞黏附、免疫学过程等生物学过程。靶基因分子功能分析显示,下调或上调的靶基因主要具有肽链内切酶抑制剂活性、肝素结合、信号受体结合等功能;信号通路预测显示TLDGD可调控PI3K-Akt信号通路、肌动蛋白细胞骨架、黏着斑等。结论:TLDGD可能通过调控多种lncRNA和mRNA的表达及下游相关信号通路来发挥治疗糖尿病肾病的作用。

Objective:To explore the mechanism of Tongluo Digui Decoction in treating diabetic kidney disease(DKD)by detecting the expression profile of long noncoding RNA(lncRNA)and messenger RNA(mRNA)in renal tissue of DKD mice.Methods:C57 BL/6 N mice were randomly divided into normal group,DKD model group and DKD+Tongluo Digui Decoction(TLDGD)group.After 8 weeks,Serum creatinine(Scr),Blood urea nitrogen(BUN),24 hProtein(24 hPro)and Fasting blood glucose(FBG)were detected;HE staining,Masson staining and PAS staining were used to observe the renal pathological changes of mice in each group;gene chip technology was used to detect the difference of lncRNA and mRNA closely related to the therapeutic effect of Tongluo Digui Decoction on DKD,and cluster analysis was carried out.Bioinformatics methods were used to predict the target genes regulated by differential lncRNA,and their functions and pathways were analyzed.Results:Compared with the normal group,Scr,BUN,24 hPro and FBG in the DKD model group were increased,and the differences were statistically significant(P<0.01).Compared with the DKD model group,the above indexes in the DKD+Tongluo Digui Decoction group were decreased,and the differences were statistically significant(P<0.05).In DKD+Tongluo Digui Decoction group,renal pathological injury was significantly improved.Tongluo Digui Decoction regulated the differential expression of 84 lncRNA,of which 42 were up-regulated and 42 were down regulated,and the differences were statistically significant(P<0.05).There were 189 differentially expressed mRNA,132 of which were up-regulated and 57 down regulated.The differences were statistically significant(P<0.05).Go analysis showed that down regulated or up-regulated target genes were mainly involved in cell migration,inflammatory response,cell adhesion,immunological process and other biological processes.The molecular function analysis of target genes showed that the down regulated or up regulated target genes mainly had the functions of endopeptidase inhibitor activity,heparin binding,signal receptor binding,etc.;the prediction of signal pathway showed that Tongluo Digui Decoction could regulate PI3 K Akt signal pathway,actin cytoskeleton,adhesive plaque,etc.Conclusion:Tongluo Digui Decoction may play a role in the treatment of DKD by regulating the expression of lncRNA and mRNA and downstream signal pathways.