IL-1β x IL-6 x TGF-β1 x TNF-α

Vascular remodeling, which results in aortic sclerosis, is one of the main risk factors for hypertension-related cardiovascular events. In the process of early intervention and clinical improvement, the inhibition of this process is very important. Previous studies have shown that liensinine (Lien) alleviates hypertension-induced vascular remodeling, but its mechanism of action remains unclear. This study aimed to investigate the therapeutic effects of Lien on hypertension, explore its molecular targets, and elucidate its anti-inflammatory mechanisms related to ferroptosis in preventing vascular remodeling. In vivo, Lien significantly reduced blood pressure and PWV and improved vascular pathology by reducing aortic wall thickness in hypertensive mice. The results of the WB experiment indicated that Lien can promote the expression of GPX4 protein and inhibit the expression of TFRC protein, which are ferroptosis markers. RNA-seq results suggested that Lien targets TLR4-mediated inflammation in vascular remodeling. The serum levels of IL-6, IL-1β, and TNF-α were decreased by Lien, according to ELISA results. IHC demonstrated decreased expression of inflammatory markers (p-IκB-α, IκB-α, p-P65, P65) in vascular tissues. In vitro, the results of the WB experiment indicated that Lien can promote the expression of GPX4 protein and inhibit the expression of TFRC protein; the ELISA results indicate that Lien can reduce the secretion of IL-6, IL-1β, and TNF-α induced by Ang II. Molecular docking and COIP experiments showed that Lien inhibits the formation of the Ang II-TLR4-MD2 complex and reduces MyD88 expression. Western blotting further confirmed that Lien inhibited the MAPK and TGF-β1/Smad2/3 pathways, both in the presence of Ang II and inflammatory stimuli (LPS, CKs). Subsequently, using the TLR4 inhibitor TAK242 and the NF-κB inhibitor PDTC, further evidence was provided that Lien effectively manages hypertension by lowering blood pressure, enhancing vascular health, and providing anti-inflammatory effects, mainly by inhibiting the TLR4/MD2 complex and suppressing the MAPK and TGF-β1/Smad2/3 pathways. These results underscore the promise of Lien as a therapeutic agent for preventing vascular remodeling caused by hypertension. KEY MESSAGES: Liensinine significantly reduces blood pressure and improves vascular inflammatory in hypertensive mice. Liensinine inhibits vascular inflammatory and remodeling by suppressing the ferroptosis related GPX4 and TLR4/MD2 complex and its downstream NF-κB, MAPK, and TGF-β1/Smad2/3 signaling pathways. Liensinine demonstrates potential as a therapeutic agent and offers new insights into inflammation-driven vascular dysfunction and its intervention.

Azoxystrobin (AZO) is a broad-spectrum fungicide that has been reported to induce severe organ toxicity in non-target organisms. The current research project explored the dose-response toxic effects of AZO on testicular tissues. Forty-eight male Sprague Dawley rats were divided into control, AZO (125 mg/kg), AZO (250 mg/kg), and AZO (500 mg/kg) treated group. AZO intoxication at all the tested doses upregulated the gene expression of TGF-β1, SMAD2, CYP19A1, FKBP5, SMAD3, COL1A1 and TGFBR1 while suppressing the expression of 3β-HSD, AR, SRD5A1, StAR, 17β-HSD, and KLK3. The levels of ROS and MDA were promoted coupled with significant reduction in the activities of HO-1, CAT, GPx, GSR, GST, and SOD after AZO administration. Besides, AZO exposure adversely altered sperm count, viability, and morphology at all the administered doses. Similarly, the levels of LH, FSH, and testosterone were suppressed following the administration of AZO. Testicular tissues showed severe pro-inflammatory response as indicated by high levels of NF-κB, TNF-α, IL-6, IL-1β, and COX-2 in response to AZO exposure. Histopathological analysis showed testicular disruptions including reduced diameter of seminiferous tubules, degeneration of Leydig cells, thinning of germinal epithelium, vascular congestion, and necrosis in dose-dependent manners of AZO exposure. Our biochemical and histological findings are further supported by computation analysis which revealed that AZO have strong affinity to bind with key regulatory genes thereby influencing their expressions.

Liver fibrosis, characterized primarily by oxidative stress and excessive extracellular matrix deposition, represents a significant health burden. TGF-β1/Smad pathway activation serves as a central driver of this process. Unfortunately, current clinical treatments for liver fibrosis all have certain limitations, highlighting the urgency of developing novel therapeutic approaches. In this study, Ganoderma lucidum polysaccharide (GLP)-MnO₂ nanoenzymes primarily treat liver fibrosis through two mechanisms: first, it exhibits potent activity of antioxidant enzymes, such as catalase (CAT) and superoxide dismutase (SOD), successfully scavenging superoxide anion (·O₂^-), hydrogen peroxide (H₂O₂), and hydroxyl radicals (·OH) in fibrotic liver regions to alleviate oxidative stress. Second, it prevents the TGF-β1/Smad pathway from being activated. Consequently, fewer Collagen I, α-SMA, and inflammatory mediators TNF-α, IL-6, and IL-1β are produced. Additionally, this nanozyme exhibits passive liver targeting and dual-modality photoacoustic imaging (PAI) and magnetic resonance imaging (MRI) properties, providing crucial support for both therapeutic intervention and disease assessment in liver fibrosis. Collectively, these results show the therapeutic potential of GLP-MnO₂ nanozyme in treating liver fibrosis.