Lingling Zhou

To assess Panax notoginseng saponins(PNS)'s therapeutic potential against RA and its modulation of Th17 differentiation, specifically the in vitro mechanism by which PNS inhibits Th17 cell glycolysis via the HIF-1α pathway. Collagen-induced arthritis (CIA) mouse model was established by immunization with type II collagen. CIA mice were randomly divided into control, model, and PNS (100 mg/kg) treatment groups. Network pharmacology was employed to identify PNS active components and RA-related targets; protein-protein interaction (PPI) network construction and pathway enrichment analysis were used to predict core targets. Naive CD4⁺T cells were induced to differentiate into Th17 cells with interleukin (IL)-6, IL-23, and transforming growth factor-β (TGF-β), and treated with PNS at concentrations of 5, 10, and 20 µg/mL). Th17 differentiation, IL-17 A secretion, and pathway-related molecules were detected by flow cytometry, ELISA, RT-qPCR, and immunoblotting. Compound 3 K and DFOM validated the mechanism. PNS attenuated CIA severity (reduced hind paw swelling, arthritis scores, improved body weight, alleviated pathology). KEGG linked Th17 differentiation, PKM2, HIF-1α to PNS's anti-RA effects. PNS reduced Th17 ratios, inhibited PKM2/HIF-1α expression, suppressed PKM2-mediated glycolysis, and reversed HIF-1α hyperactivation in Th17 cells. PNS relieves CIA by inhibiting Th17 differentiation via suppressing HIF-1α-mediated PKM2 expression and glycolysis-dependent Th17 differentiation.

Purpose To assess the therapeutic potential of PNS against rheumatoid arthritis (RA) and its modulation of Th17 differentiation, specifically investigating in vitro the mechanism by which PNS inhibits glycolysis in Th17 cells through the HIF-1α signaling pathway. Methods CIA model were established and allocated into control, model, and PNS (100 mg/kg) groups to assess anti-arthritic efficacy and effects on Th17 differentiation. Network pharmacology identified PNS components and RA-related targets across databases. Protein-protein interaction (PPI) network construction and pathway enrichment analysis predicted core targets and pathways. Naive CD4⁺ T cells were differentiated into Th17 cells using IL-6, IL-23, and TGF-β. Cells (excluding the control group) were treated with PNS (5, 10, 20 µg/mL). Th17 cell differentiation was assessed by flow cytometry, and IL-17A secretion was quantified by ELISA. Expression of pathway-related genes and proteins was determined via RT-qPCR and immunoblotting, respectively. The PKM2-specific inhibitor Compound 3K and HIF-1α activator DFOM were employed to validate the mechanistic pathway. Results PNS attenuated the severity of arthritis in CIA mice, which was evidenced by the swelling of the hind paws, arthritis scores, body weight and pathological changes. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that Th17 cell differentiation, PKM2 and HIF-1α signaling pathways may be related to the anti-RA effect of PNS. PNS effectively reduced the ratio of Th17 cells in vitro and in vivo. PNS inhibited the expression of PKM2 and HIF-1α in synovial cells and spleen T cells of CIA mice. PNS treatment significantly inhibited PKM2-mediated glycolysis. Furthermore, hyperactivation of the HIF-1α pathway in Th17 cells was reversed by PNS treatment. Conclusions PNS relieved the severity of CIA mice by inhibiting Th17 differentiation. Mechanistically, PNS suppressed HIF-1α-mediated PKM2 expression, inhibiting glycolysis-dependent Th17 differentiation., thereby alleviating rheumatoid arthritis.

Triptolide (TP), known for its effectiveness in treating various rheumatoid diseases, is also associated with significant hepatotoxicity risks. This study explored Catalpol (CAT), an iridoid glycoside with antioxidative and anti-inflammatory effects, as a potential defense against TP-induced liver damage. In vivo and in vitro models of liver injury were established using TP in combination with different concentrations of CAT. Metabolomics analyses were conducted to assess energy metabolism in mouse livers. Additionally, a Seahorse XF Analyzer was employed to measure glycolysis rate, mitochondrial respiratory functionality, and real-time ATP generation rate in AML12 cells. The study also examined the expression of proteins related to glycogenolysis and gluconeogenesis. Using both in vitro SIRT1 knockout/overexpression and in vivo liver-specific SIRT1 knockout models, we confirmed SIRT1 as a mechanism of action for CAT. Our findings revealed that CAT could alleviate TP-induced liver injury by activating SIRT1, which inhibited lysine acetylation of hypoxia-inducible factor-1α (HIF-1α), thereby restoring the balance between glycolysis and oxidative phosphorylation. This action improved mitochondrial dysfunction and reduced glucose metabolism disorder and oxidative stress caused by TP. Taken together, these insights unveil a hitherto undocumented mechanism by which CAT ameliorates TP-induced liver injury, positioning it as a potential therapeutic agent for managing TP-induced hepatotoxicity.

Background: Rheumatoid arthritis (RA) is a persistent and systemic autoimmunity disease. The abnormal differentiation of Treg cells is important in pathogenesis. Despite previous studies showed that microRNAs (miRNAs, miR) are pivotal modulators of Treg cells, the effect of miRNAs on Treg cell differentiation and function is not clear. Our study wants to reveal the relationship of miR‐143‐3p with the differentiative ability and biofunction of Treg cells during the development of RA.Methods: The Expressing level of miR‐143‐3p and cell factor generation in peripheral blood (PB) of RA sufferers were identified by ELISA or RT‐qPCR. The roles of miR‐143‐3p in Treg cell differentiation were studied via ShRNA/lentivirus transfection. Male DBA/1 J mice were separated into control, model, control mimics, and miR‐143‐3p mimics groups to analyze the anti‐arthritis efficacy, the differentiative ability of Treg cells, and the expression level of miR‐143‐3p.Results: Our team discovered that the Expressing level of miR‐143‐3p was related to RA disease activities in a negative manner, and remarkably related to antiinflammation cell factor IL‐10. In vitro, the expression of miR‐143‐3p in the CD4⁺T cells upregulated the percentage of CD4⁺CD25⁺Fxop3⁺ cells (Tregs) and forkhead box protein 3 (Foxp3) mRNA expression. Evidently, miR‐143‐3p mimic intervention considerably upregulated the content of Treg cells in vivo, validly avoided CIA progression, and remarkably suppressed the inflammatory events of joints in mice.Conclusion: Our findings indicated that miR‐143‐3p could ameliorate CIA through polarizing naive CD4⁺T cells into Treg cells, which may be a novel strategy to treat autoimmune diseases such as RA.

Rheumatoid arthritis (RA) is an autoimmune disease with aberrant Th17 cell differentiation. Panax notoginseng (Burk.) F. H. Chen (Araliaceae) saponins (PNS) have an anti-inflammatory effect and can...