Tinosinenside A inhibits neuroinflammation and protects HT22 cells by suppressing the TLR4/NF-κB/NLRP3 signaling pathway in BV2 cells

Microglia-mediated neuroinflammation plays a crucial role in Alzheimer's disease (AD). Tinosinenside A (Tis A) is a novel sesquiterpene glycoside isolated from the dried rattan stem of Tinospora sinensis (Lour.) Merr. Tis A exhibited anti-inflammatory and neuroprotective activities in vitro. However, the mechanism underlying the inhibition of neuroinflammation and protection of nerve cells remains obscure. This study used lipopolysaccharide (LPS)-induced inflammatory response in BV2 cells to simulate a neuroinflammatory model and used Aβ_1-42-induced HT22 cells to establish an AD cell model, aiming to investigate the efficacy and mechanism of Tis A through anti-neuroinflammation to protect nerve cells. Tis A had no effect on the proliferation of BV2 and HT22 cells at the tested concentrations. The time- and dose-dependent effects of Tis A on the LPS-induced inflammatory response of BV2 cells demonstrated that the best anti-inflammatory efficacy appeared after 12 h of pretreatment. Tis A inhibited the gene levels of TNF-α, IL-6, IL-1β, iNOS, and IL-10 while enhancing the gene levels of IL-4 and TGF-β. Additionally, Tis A reduced the gene expression levels of CD16 and CD32 and increased the CD36 and CD206 gene expression levels. It also downregulated the protein expression of Iba-1 and iNOS while upregulating CD206. Tis A obviously inhibited NLRP3 gene and protein expression in LPS-stimulated BV2 cells. The inhibitory effect of Tis A on NLRP3 was counteracted by the NLRP3 Activator nigericin and overexpression plasmid GV358. Tis A inhibits NLRP3 protein expression to reduce the assembly of NLRP3/ASC/Caspase-1 inflammasome, then regulates the TLR4/NF-κB/NLRP3 signaling pathway. It regulates microglia activation and M1/M2 phenotypic polarization, then inhibits the production of inflammatory factors, and reduces the Apoptosis rate of HT22 cells under inflammatory conditions, improving the survival rate of nerve cells to protect neurons.