Comparative Study On The Inhibitory Effects Of GLP-1/GIP Dual Receptor Agonist DA-CH5 And GLP-1 Single Receptor Agonist NLY01 On NF-κB Inflammation Pathway In MPTP Mouse Model

Objective:To study and compare the protective effect of the new GLP-1/GIP dual-receptor agonist DA-CH5(Also known as DA5-CH or DA5)and the single-receptor agonist NLY01 on the NF-κB neuroinflammatory pathway in the PD model of MPTP mice and its specific mechanism.Method:MPTP was injected intraperitoneally to induce a mouse Parkinson’s disease-like model.72 8-week-old,healthy male C57BL/6 mice were randomly divided into four groups: C group(Control group)and M group(MPTP model group),MD group(MPTP+DA5 group),MN group(MPTP+NLY01 group),the sample size of each group is 18.The PD model was induced by intraperitoneal injection of MPTP(25mg/kg/d).Mice in MD and MN groups were injected intraperitoneally with DA-CH5 and NLY01(25nmol/kg/d)half an hour after MPTP injection.Mice in MD and MN groups were injected intraperitoneally with DA-CH5 and NLY01(25nmol/kg/d).Give the same volume of saline.After 7 days,the brain was sacrificed for perfusion and the brain was taken for follow-up testing.Behavioral tests include observing the spontaneous behavior,gait,open field and rotating rod experiments of mice.The following indicators were detected by Western Blot and IHC: characteristic pathological indicators of Parkinson’s disease: tyrosine hydroxylase(TH),α-synuclein(α-Syn);TLR4/NF-κ B pathway indicators: Toll-like receptors Body-4(TLR4),nuclear factor-κB(NF-κB)and tumor necrosis factor(TNF-α);glial cell-related inflammation indicators: microglial cell activation specific representative markers(IBa-1),star Representative markers of glial cell activation-glial fibrillary acidic protein(GFAP)and transforming growth factor β1(TGF-β 1);neurotrophic factor: glial cell-derived neurotrophic factor(GDNF),brain-derived nerve The expression of trophic factor(BDNF);ELisa was used to detect the expression of neuroinflammation-related factors IL-10,IL-6 and IL-Iβ.Result:The results of behavioral experiments showed that the gait behavior of the MD group mice was significantly improved(P<0.0001),while only some gait indicators of the MN group were improved.Both drugs restored the expression of neuron tyrosine hydroxylase in the substantia nigra and striatum(P<0.05),but NLY01 showed no improvement on α-synuclein(P>0.05).More importantly,DA-CH5 played a better role in improving glial cell activation and related markers of NF-κB inflammation pathway(P<0.05).Conclusion:The results show that DA-CH5 can well improve the behavioral indicators of MPTP mice,and may play a neuroprotective effect on MPTP mice through the TLR4/NF-κB neuroinflammatory signaling pathway,and the effect is better than NLY01.