背景:探讨川陈皮素对高脂饮食(HFD)所致肠神经损伤的保护作用及其机制。
方法:通过HFD诱导肥胖模型。通过灌胃给药诺美林(100mg/kg和200mg/kg)和媒介物4周。李的索引,体重,在处死前进行OGTT和肠推进测定。取样后,使用Bodipy493/503检测脂质;使用MDA和SOD试剂盒检测脂质过氧化以及PGP9.5,Trem2,GFAP,β-微管蛋白3,Bax,Bcl2,Nestin,P75NTR,使用免疫荧光检测SOX10和EDU。GDNF,p-AKT,AKT,p-FOXO3a,使用蛋白质印迹法检测FOXO3a和P21蛋白。通过qPCR检测NOS2的相对mRNA表达水平。培养原代肠神经干细胞(ENSC)。ENSC用棕榈酸(PA)和川陈皮素处理后,进行CCK-8和半胱天冬酶-3/7活性测定以评估增殖和凋亡。
结果:HFD消耗导致结肠脂质积累和过氧化,引起肠神经损伤并引起肠运动功能障碍。然而,nobiletin减少脂质积累和过氧化在结肠;促进Trem2,β-微管蛋白3,巢蛋白,P75NTR,SOX10和Bcl2表达;抑制Bax和GFAP表达;降低NOS2mRNA转录;并调节GDNF/AKT/FOXO3a/P21途径。Nobiletin还促进PA诱导的ENSC损伤。
结论:诺比林可恢复HFD引起的肠神经损伤,这可能与抑制肠神经细胞凋亡有关,促进肠神经存活和调节GDNF/AKT/FOXO3a/P21通路。
BACKGROUND: To explore whether nobiletin has a protective effect on high-fat diet (HFD)-induced enteric nerve injury and its underlying mechanism.
METHODS: An obesity model was induced by a HFD. Nobiletin (100 mg/kg and 200 mg/kg) and vehicle were administered by gastric gavage for 4 weeks. Lee\'s index, body weight, OGTT and intestinal propulsion assays were performed before sacrifice. After sampling, lipids were detected using Bodipy 493/503; lipid peroxidation was detected using MDA and SOD kits and the expression of PGP 9.5, Trem2, GFAP, β-tubulin 3, Bax, Bcl2, Nestin, P75 NTR, SOX10 and EDU was detected using immunofluorescence. The GDNF, p-AKT, AKT, p-FOXO3a, FOXO3a and P21 proteins were detected using western blotting. The relative mRNA expression levels of NOS2 were detected via qPCR. Primary enteric neural stem cells (ENSCs) were cultured. After ENSCs were treated with palmitic acid (PA) and nobiletin, CCK-8 and caspase-3/7 activity assays were performed to evaluate proliferation and apoptosis.
RESULTS: HFD consumption caused colon lipid accumulation and peroxidation, induced enteric nerve damage and caused intestinal motor dysfunction. However, nobiletin reduced lipid accumulation and peroxidation in the colon; promoted Trem2, β-tubulin 3, Nestin, P75NTR, SOX10 and Bcl2 expression; inhibited Bax and GFAP expression; reduced NOS2 mRNA transcription; and regulated the GDNF/AKT/FOXO3a/P21 pathway. Nobiletin also promoted PA-induced impairment of ENSCs.
CONCLUSIONS: Nobiletin restored HFD-induced enteric nerve injury, which may be associated with inhibiting enteric nerve apoptosis, promoting enteric nerve survival and regulating the GDNF/AKT/FOXO3a/P21 pathway.