Clinical Significance Of AKR1B10 And SPP1 In Nonalcoholic Fatty Liver Disease

AIM:Non-alcoholic fatty liver disease(NAFLD),which includes two different pathological status,simple fatty liver(NAFL)and nonalcoholic steatohepatitis(NASH).In recent years,the incidence of NAFLD has been increasing,but the underlying pathogenesis of NAFLD is still obscure.In this study,gene expression profiles of NAFLD patients were analyzed to find genes that are related to the progression and prognosis of NAFLD,to screen therapeutic sensitive drugs and conduct experimental verification.Methods:In the first part,we downloaded NAFLD and HCC gene expression profiles and clinical data from public databases.Subsequently,NAFLD-related genes were obtained by intersecting the differentially expressed genes of two groups(NAFL-NASH group;Normal-NASH group).Then,these differential genes were intersected with traits-related genes obtained by WGCNA algorithm to obtain NAFLD progression-related genes.NAFLD progression related genes were analyzed for survival,and Normal-NAFL-NASH-HCC progression genes,AKR1B10 and SPP1 were obtained.Later,TGX-221,which is sensitive to the treatment of NAFLD,was screened from GDSC database.In the second part,H&E staining was used to observe the pathological changes of liver tissue,oil red staining was used to observe the lipid deposition,Sirius red staining was used to show the accumulation of liver collagen,immunohistochemistry and QPCR were used to verify the protein and gene expression.Results:The first partIn this part,we established three disease status classifiers to distinguish the disease status of patients with different NAFLD.Secondly,we obtained 2 progression-related genes(AKR1B10 and SPP1),and theses gene expression level were closely related to patient survival(P<0.05).In addition,TGX-221 was screened for the treatment of NAFLD.The second part:1.Mice H&E staining:No abnormality was found in the normal group.Mice in the MCD 2-week group(MCD-2W)showed mild steatosis.Steatosis was further aggravated in the 4-week group(MCD-4W).The 8-week group(MCD-8W)presented severe fatty liver and partial fibrosis,which showed NASH state.The TGX-221 group presented mild to moderate steatosis.2.Mice Sirius Scarlet Staining:The normal group contained only a small amount of collagen in the vessel wall.Collagen increased gradually in MCD-2W mice;Collagen in MCD-4W mice mainly existed in portal area and surrounding area.The collagen of MCD-8W mice increased further and had a tendency to form pseudoblobule.Content of collagen in TGX-221 group was roughly between MCD-4W and MCD-8 W mice.3.QPCR:AKR1B10 and SPP1 expressions in normal group and MCD(2W,4W,8W)mice showed an upward trend.The expression of AKR1B10 and SPP1 in TGX-221 treatment group was lower than that in MCD-8W mice.4.Immunohistochemistry staining of mice:AKR1B10 and SPP1 were almost not expressed in the normal group,while only a small amount of AKR1B10 and SPP1 were expressed in MCD-2W mice.At the beginning of 4 weeks,the staining was further deepened,and at 8 weeks,the staining was the deepest.The staining of TGX-221 treatment group was similar to that of MCD-4W mice.5.Staining of patient tissue:According to H&E staining and immunohistochemistry of NAFLD patients,pseudolobular formation and inflammatory cell infiltration were shown.Sirius-red staining showed a tendency of pseudolobular formation,suggesting that the patient was in NASH status.Immunohistochemistry showed that AKR1B10 and SPP1 were highly expressed in NASH patients.Conclusions:1.This study established 3 disease status classifiers,which can be used to distinguish different NAFLD disease states;2.AKR1B10 and SPP1 is related to NAFLD disease progression;3.TGX-221 can be used as a potential molecular target for the treatment of NAFLD in the future.