| Objective:To study the effect of Shenqi compound on diabetic thoracic aortic disease in GK rats,established a diabetic thoracic aortic disease animal model with GK rats.obtain the mRNA/lncRNA expression profile in the horacic aortic of GK rats through deep sequencing,further screen the co-aggregation gene which is both differential mRNA and differential lncRNA target gene,and perform bioinformatics analysis via GO and KEGG enrichment,to explore possible molecular mechanisms of Shenqi compound for diabetic thoracic aortic disease.Methods:The first part: Effect of Shenqi Compound on Thoracic Aortic Disease in GK Rats.100 male GK rats with fasting blood glucose ?11.1mmol/L at7-8 weeks of age were fed ad libitum for one week(7 days),and then fed with high-fat diet for 4 weeks to simulate metabolic memory process to establish diabetic macroangiopathy(thoracic aorta)model.After successful modeling,they were randomly divided into model group(M group),western medicine group(W group),Shenqi compound high-dose group(ZH group),Shenqi compound middle-dose group(ZM group),and Shenqi compound low-dose group(ZL group),20 rats in each group,and another 20 Wistar rats were set as a control group(group C)with the ordinary feed.Each group was given the corresponding drugs,and the model group and the normal group were given distilled water for a period of 12 weeks(84 days).The general condition of all rats was observed during the experiment.The fasting plasma sugar level of the rats was monitored weekly.After the intervention,the levels of pro-inflammatory factor IFN-? and anti-inflammatory factor IL-4 were measured and the IFN-?/IL-4 was calculated.The pathological changes of thoracic aorta in each group were observed under HE staining.The thickness of the vessel wall was calculated.The apoptosis of thoracic aorta in each group was observed by Tunel apoptosis staining,and the apoptosis rate was calculated.The second part: Molecular mechanism of Shenqi compound on thoracic aortic lesions in GK rats.The modeling and intervention methods are the same as the first part.After the intervention,4 rats ineach group were randomly selected to obtain the mRNA/lncRNA expression profile in the horacic aortic of GK rats through deep sequencing,further screen the co-aggregation gene which is both differential mRNA and differential lncRNA target gene,and perform bioinformatics analysis via GO and KEGG enrichment,to explore possible molecular mechanisms of Shenqi compound for diabetic thoracic aortic disease.Results:The first part: 1.Successfully constructed a model of thoracic aortic disease in GK rats: The diabetic thoracic aortic disease rat model was successfully constructed and characterized as:Comparing with the control group,The fasting plasma glucose of diabetic thoracic aortic disease rats significantly exceeded the normal group,general condition is very poor,slow weight gain,and there were risk factors-persistent hyperglycemia,direct pathological observation of thoracic aorta It showed severe endothelial damage,increased number of apoptosis,increased apoptosis rate,independent risk factor-inflammatory mediator disorder(pro-inflammatory factor IFN-? level increased,anti-inflammatory factor IL-4 level decreased,IFN-? /IL-4 ratio increased).2.Shenqi compound has a good effect on thoracic aortic disease in GK rats:After the treatment of Shenqi compound,Comparing with the model group,the mental state of Shenqi compound group gradually recovered,fasting plasma glucose showed a downward trend,proinflammatory factor IFN-? level showed a downward trend,anti-inflammatory The level of factor IL-4 is increased,the ratio of IFN-? /IL-4 is decreased,the endothelial damage of thoracic aorta is restored to normal,the apoptosis is decreased,and the apoptosis rate is significantly reduced.So,From multiple aspects including risk factors of diabetic thoracic aorta,pathophysiological mechanism and inflammatory medium,reflects the intervention effect of Shenqi compound on diabetic thoracic aortic disease.The second part: 1,Shenqi compound can widely regulate the abnormal expression of mRNA: There were differences in mRNA expression levels between the diabetic thoracic aortic disease model and the normal control group determined the difference in pathological conditions of diabetic thoracic aortic disease;After the treatment of Shenqi compound,the abnormal expression of differential mRNA was adjusted to a certain extent.the differential mRNAs in the first ten reversible regulation were: RGD1304770,Nppa,Mpz,Mx2,Prss8,Hoxc10,Hoxc9,Best3,Casq1,Rgs1,Otog,Plch1,Cish,Nrg2,AC141997.1,Fam196 a,AABR07050487.1,Scd1,Fmo3.2.Shenqi compound can reverse the abnormal expression of Tp53inp1,Gadd45 b,Adrb2,Myc,Ccl21 related to inflammatory response and apoptosis through cAMP/cGMP-PKG signaling pathway: GO analysis shows that Shenqi compound can be reversely regulated and associated with inflammatory response and apoptosis and the differential mRNAs including Tp53inp1,Gadd45 b,Adrb2,Myc,Ccl21.The Bioinformatic analysis showed these differential mRNAs may be involved in multiple signaling pathway such as cAMP signaling pathway,calcium signaling pathway,cGMP-PKG signaling pathway,parotid secretion,proteoglycan The role of cancer(P<0.05).3.Shenqi compound can widely regulate the abnormal expression of lncRNA: There were differences in lncRNA expression levels between the diabetic thoracic aortic disease model and the normal control group determined the difference in pathological conditions of diabetic thoracic aortic disease;After the treatment of Shenqi compound,the abnormal expression of differential lncRNA was adjusted to a certain extent.the differential lncRNAs in the first ten reversible regulation were: Gtf3c1.gSep08,Stat3.eSep08 and Etfb.cSep08 et al.4.Shenqi compound can regulate the abnormal expression of the common gene Rgs2,Tp53inp1,Best3,Gprin3 and Aplnr:Compared with the normal group,There were differential expression of differential mRNA and the differential lncRNA target gene expression levels between the diabetic thoracic aortic disease model and the normal control group determined the difference in pathological conditions of diabetic thoracic aortic disease;The Shenqi compound could regulate the abnormal expression,and the Shenqi compound could reversely regulate the gene expression of Rgs2,Tp53inp1,Best3,Gprin3.Shenqi compound high,medium and low dose can regulate gene expression of Aplnr.5.Shenqi compound can regulate the expression of differential mRNA/lncRNA related to inflammation and apoptosis: GO analysis found that the differential mRNA,differential lncRNA and its co-assembledgenes regulated by Shenqi compound are involved in the biological processes including inflammatory response and apoptosis.6.The signaling pathways can be regulated by Shenqi compound including fatty acid metabolism,fatty acid degradation and cGMP-PKG: The model group is compared with the control group displays that Bioinformatic analysis showed these differences mRNAs and differential lncRNAs target gene co-assembled genes may participate in many signaling pathways,The top five in the passway are closely related to fatty acid metabolism and fatty acid degradation.Compared with the model group,the high-dose group of Shenqi compound was also closely related to fatty acid metabolism and fatty acid degradation in the top five of the passway;In addition,these differentially co-assembled genes regulated by the high,middle and low dose groups of Shenqi compound are closely related to the cGMP-PKG signaling pathway.7.The co-expression analysis of differential lncRNA and differential mRNA showed that the core regulatory lncRNA of diabetic thoracic aortic lesions was lncRNA:Gtf3c1.gSep08,regulated by Shenqi compound in the previous results of Nrep.bSep08,Col5a1.aSep08,soygee.aSep08-unspliced,NONRATT013247.2,votar.aS ep08-unsplice,.Stat3.eSep08,Etfb.cSep08 and differential mR NA showed strong correlation,which further expounded that Shenqi compound could regulate the abnormal expression of differential lncRNA.Conclusion:1.TIt is feasible to use GK rats with high-fat continuous feeding to perform modeling of diabetic thoracic aortic disease.2.The core regulation of lncRNA in diabetic rats with thoracic aortic disease areNrep.bSep08,Col5a1.aSep08,soygee.aSep08-unspliced,NONRATT013247.2,votar.aSep08-unsplice.3.Shenqi compound is effective in the treatment of diabetic thoracic aortic disease.And the possible mechanism is that Shenqi compound can regulate the mRNA and lncRNA,which are closely related to inflammatory reaction and apoptosis process,is widely regulated in order to treat the cocollection geneRgs2,Tp53inp1,Best3,.The regulation of Gprin3,Aplnr and differential lncRNA:Gtf3c1.gSep08,Stat3.eSep08,Etfb.cSep08 is the most obvious.Which the involved signal pathway including fatty acid metabolism,fatty acid degradation and cGMP-PKG signal pathway.Based on the theory of accumulation of evil,combined with the pathogenesis characteristics of "deficiency and turbid heat toxin" in diabetic macrovascular(thoracic aortic)lesions,it reflects the advantages of Shenqi compound inhibite the fuzheng evil,and inhibe metabolic memory,and the ability of comprehensively regulating of Chinese medicine. |
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