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Ulinastatin inhibits NLRP3-induced apoptosis in a PD cell model.

Annals of translational medicine (2021-08-06)
Yicheng Lin, Dongsheng Xu, Feng Gao, Xiaolei Zheng
ABSTRACT

Emerging evidence suggests that inflammation induced by the inflammasome plays a crucial role in the course of Parkinson's disease (PD). Ulinastatin (UTI) has shown significant anti-inflammatory effects. However, few studies have examined whether UTI protects neurons through its anti-inflammatory effects in PD. The purpose of this study is to determine whether UTI exerts neuroprotection in a PD cell model and to explore the mechanisms. SH-SY5Y cells and nerve growth factor (NGF)-treated PC12 cells were used to establish MPP+ induced PD cell models. Cells were pre-treated with UTI, then cell viabilities were detected using the MTT assay. Lactate dehydrogenase (LDH) release was detected using the LDH release assay kit. Inflammatory factors such as IL-1β, IL-6, and TNF-α were detected using ELISA. The expression levels of TH, NLRP3, caspase-1, ASC, IL-1β, and IL-18 were measured using western blotting, and DA release was detected using HPLC. A NLRP3 activator Nigericin was used to verify the effect of NLRP3 in the neuroprotective mechanism of UTI. We observed decreased cell viability, increased apoptosis, and increased inflammatory factors such as IL-1β, IL-6, and TNF-α in the MPP+ induced PD model. We also found decreased DA secretion and TH expression, as well as increased NLRP3, caspase-1, ASC, IL-1α, and IL-18 expression in the MPP+ induced PD model. These changes were significantly attenuated by UTI pre-treatment in a dose dependent manner. NLRP3 activator Nigericin markedly increased LDH release, accelerated apoptosis, increased inflammation levels and decreased DA secretion and TH expression, suggesting that Nigericin eliminated the neuroprotective effect of UTI on MPP+ treated cells. Our data demonstrated that UTI pre-treatment performed a neuroprotective role in the MPP+ induced PD cell models by inhibiting the NLRP3 pathway.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-GAPDH antibody produced in rabbit, ~1 mg/mL, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
Anti-NLRP3 antibody produced in rabbit, purified immunoglobulin, buffered aqueous solution
Sigma-Aldrich
Anti-Caspase-1 (ab1) antibody produced in rabbit, affinity isolated antibody, buffered aqueous solution