Role of WNT1-induced signaling pathway protein 1 in non-alcoholic steatohepatitisand underlying mech-anism
Objective To explore the functional localization and therapeutic mechanism of WNT1-in-duced signaling pathway protein 1(WISP1)in non-alcoholic steatohepatitis(NASH).Methods A total of 60 male C57BL/6J mice were divided into 5 groups by simple random sampling(10 mice in each group).The normal group was fed on ordinary diet,and the NASH mouse model was prepared by feeding high-fat and high-cholesterol diet for 14 weeks.During the modeling process,the mice with intraperitoneal injection of IgG mono-clonal antibody,0.1 g/kg WISP-1 high-affinity monoclonal antibody,0.2 g/kg WISP-1 high-affinity monoclonal antibodies served as IgG group,low-dose WISP-1 antibody group and high-dose WISP1 antibody group,respec-tively.All mice in three groups were administered twice a week for 14 weeks.Serum alanine aminotransferase(ALT),aspartate aminotransferase(AST)and blood glucose levels were detected.The pathological changes of liver tissue were observed by hematoxylin-eosin(HE),glycogen(PAS)and oil red O staining.The levels of to-tal cholesterol(TC)and triglycerides(TG)in liver tissue were detected,and the activity levels of reduced glu-tathione(GSH),malondialdehyde(MDA)and superoxide dismutase(SOD)were detected by enzyme-linked im-munosorbent assay(ELISA).The mRNA expression levels of interleukin(IL)-10,IL-6,IL-1β and tumor necro-sis factor-α(TNF-α)in serum of mice were detected by real-time quantitative polymerase chain reaction(Real-time PCR).The expression of WISP1 and Acetyl-CoA synthetase(ACS)in liver tissues was detected by immu-nohistochemistry(IHC),and the expression levels of adenosine monophosphate-actived protein kinase(AMPK),phosphorylated AMPK(p-AMPK),carnitine palmitoyltransferase(CPT1)and peroxisome proliferator-activated receptor-gamma coactivator(PGC)-1alpha(PCG-1α)in liver tissues were detected by Western blotting.Results Compared with the normal group,the serum AST,ALT and blood glucose levels in the model group were in-creased[(245.01±57.57)U/L vs.(37.28±4.23)U/L,(218.96±55.27)U/L vs.(49.81±10.57)U/L,(25.73±2.29)mmol/L vs.(8.06±0.51)mmol/L,P＜0.01].The pathological manifestations included hepa-tocyte steatosis,inflammatory cell infiltration and multiple necrotic foci.The mRNA levels of hepatic inflamma-tory factors(IL-10,IL-6,IL-1β,TNF-α)were increased(7.49±059 vs.1.00±0.38,5.57±3.22vs.1.00± 0.59,4.86±3.86 vs.1.00±0.34,10.18±3.24 vs.1.00±0.17,P＜0.01).The contents of MDA increased[(3.75±0.40)vs.(2.22±0.24)nmol/ml,(P＜0.01],and GSH and the activity of SOD decreased[(0.22± 0.06)vs.(0.49±0.15)ng/ml,(2.19±0.23)vs.(4.59±0.22)pg/ml,P＜0.01].The ROS content was higher than that of normal group(4.91±0.31 vs.1.00±0.12,P＜0.01).At the same time,the expression of PGC-1α,ACS protein decreased[(2.19±0.084 vs.1.00±0.09,(62.97±2.38)％vs.(26.72±0.73)％,P＜0.01],and the expression of CPT1 increased(0.55±0.01 vs.1.00±0.132,P＜0.01).Compared with the model group,the serum AST,ALT and blood glucose levels in low and high dose groups were decreased[(65.53±25.85),(53.49±12.22)U/L vs.(245.01±57.57)U/L,(73.93±23.670),(60.88±23.38)U/L vs.(218.96±55.27)U/L,(20.07±2.03),(16.46±3.71)mmol/L vs.(25.73±2.29)mmol/L,P＜0.01),the degree of liver steatosis,inflammatory cell infiltration and necrosis was decreased,and the contents of IL-10,IL-6,IL-1β,TNF-α mRNA(2.98±0.31,2.579±0.22 vs.7.49±1.57,1.83±0.35,2.53±0.32 vs.5.57±1.03,1.62±0.24,1.52±0.34 vs.4.86±1.33,2.63±0.56,2.12±0.37 vs.10.18±2.31,P＜0.05 P＜0.01),and MDA in liver tissue were significantly decreased[(1.83±0.694)vs.(3.70±0.50)nmol/ml,(P＜0.01].GSH and SOD activity was significantly increased[(0.45±0.12)vs.(0.22±0.06)ng/ml,(5.50±0.98)vs.(2.19±0.40)pg/ml,P＜0.01].The expression of PGC-1α and ACS protein was signif-icantly decreased[1.22±0.16,1.07±0.05 vs.2.19±0.08;(33.50±3.45)％,(35.38±3.65)％vs.(62.97±2.38)％,P＜0.01],and the expression of p-AMPK and CPT1 was significantly increased(2.61±0.363,2.47±0.36 vs.1.04±0.14;1.23±0.23,1.27±0.03 vs.0.55±0.01,P＜0.01).Conclusion Anti-WISP1 therapy can improve liver lipid metabolism,reduce inflammation and lipid per-oxidation,and promote liver cell repair in NASH mice by activating AMPK pathway and mediating PGC-1α/CPT1 pathway.
WNT1 induced signaling pathway protein 1Non-alcoholic SteatohepatitisLipid metabolismAdenosine monophosphate-actived protein kinase pathwayLipid peroxidation
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