Integrated analysis reveals crosstalk between pyroptosis and immune regulation in renal fibrosis.

Purpose: The pathogenesis of renal fibrosis (RF) involves intricate interactions between profibrotic processes and immune responses. This study aimed to explore the potential involvement of the pyroptosis signaling pathway in immune microenvironment regulation within the context of RF. Through comprehensive bioinformatics analysis and experimental validation, we investigated the influence of pyroptosis on the immune landscape in RF. Methods: We obtained RNA-seq datasets from Gene Expression Omnibus (GEO) databases and identified Pyroptosis-Associated Regulators (PARs) through literature reviews. Systematic evaluation of alterations in 27 PARs was performed in RF and normal kidney samples, followed by relevant functional analyses. Unsupervised cluster analysis revealed distinct pyroptosis modification patterns. Using single-sample gene set enrichment analysis (ssGSEA), we examined the correlation between pyroptosis and immune infiltration. Hub regulators were identified via weighted gene coexpression network analysis (WGCNA) and further validated in a single-cell RNA-seq dataset. We also established a unilateral ureteral obstruction-induced RF mouse model to verify the expression of key regulators at the mRNA and protein levels. Results: Our comprehensive analysis revealed altered expression of 19 PARs in RF samples compared to normal samples. Five hub regulators, namely PYCARD, CASP1, AIM2, NOD2, and CASP9, exhibited potential as biomarkers for RF. Based on these regulators, a classifier capable of distinguishing normal samples from RF samples was developed. Furthermore, we identified correlations between immune features and PARs expression, with PYCARD positively associated with regulatory T cells abundance in fibrotic tissues. Unsupervised clustering of RF samples yielded two distinct subtypes (Subtype A and Subtype B), with Subtype B characterized by active immune responses against RF. Subsequent WGCNA analysis identified PYCARD, CASP1, and NOD2 as hub PARs in the pyroptosis modification patterns. Single-cell level validation confirmed PYCARD expression in myofibroblasts, implicating its significance in the stress response of myofibroblasts to injury. In vivo experimental validation further demonstrated elevated PYCARD expression in RF, accompanied by infiltration of Foxp3+ regulatory T cells. Conclusions: Our findings suggest that pyroptosis plays a pivotal role in orchestrating the immune microenvironment of RF. This study provides valuable insights into the pathogenesis of RF and highlights potential targets for future therapeutic interventions.

Synthesis, characterization and in vitro evaluation of a series of novel polyrotaxane-based delivery system for artesunate.

A series of novel artesunate-polyrotaxanes (ATS-PRs) with folic acid capped, in which artesunate (ATS) was covalently bound to a cyclodextrin (CD) of the polyrotaxane (PR), were synthesized and were characterized by NMR, XRD, TG and DSC. The cytotoxicities of ATS-PRs on human colon cancer cell lines HT-29, SW480, HTC116 and DLD-1 showed that their antitumor activities were better than that of artesunate (ATS) and dihydroartemisinin (DHA). These ATS-PRs may provide a useful approach to the development of a highly effective drug candidate for the chemotherapy of human colon cancer.

Synthesis, characterization, and in vitro evaluation of artesunate-ß-cyclodextrin conjugates as novel anti-cancer prodrugs.

A novel series of artesunate-ß-cyclodextrin (ATS-ß-CD) conjugates, in which artesunate (ATS) was coupled covalently to one of the primary hydroxyl groups of ß-cyclodextrin (ß-CD) through amino bond formation, were synthesized and characterized by (1)H NMR, HRMS, 2D NMR (ROESY), X-ray diffraction (XRD), and thermogravimetric analysis (TGA). The results showed that the aqueous solubility of ATS-ß-CD conjugates was 26-45 times better than that of free ATS. The cytotoxicity of the ATS-ß-CD conjugates was evaluated on human colon cancer cell lines HCT116, LOVO, SW480, and HT-29, and the results indicated that ATS-2NßCD exhibited a very high cytotoxicity against HCT116, LOVO, and HT-29 with IC50 values of 0.58, 1.62, and 5.18µmol/L, respectively. In addition, the supposition of better cytotoxicity was further supported by the control experiment of fluorescent cyclodextrin.

Host-guest inclusion system of norathyriol with ß-cyclodextrin and its derivatives: preparation, characterization, and anticancer activity.

The characterization, binding ability and inclusion complexation behavior of the inclusion complexes of norathyriol with ß-cyclodextrin (ß-CD) and its derivatives such as hydroxypropyl-ß-cyclodextrin (HPßCD), sulfobutyl ether ß-cyclodextrin (SBEßCD) and mono (6-ethylene-diamino-6-deoxy)-ß-cyclodextrin (ENßCD) were investigated in both solution and solid state by means of femtosecond spectroscopy, (1)H and 2D nuclear magnetic resonance, powder X-ray diffraction. The results showed that the aqueous solubility of the complexes was much higher than that of norathyriol. The cytotoxicity of complexes on human colon cancer cell lines HT-29, SW480, Lovo and HCT116 indicated that the antitumor activities of the complexes were better than that of norathyriol. This high antitumor activity, along with the satisfactory aqueous solubility of the complexes, will be potentially useful for their application on cancer chemotherapies.

Host-guest inclusion system of scutellarein with 2-hydroxypropyl-beta-cyclodextrin: preparation, characterization, and anticancer activity.

The inclusion complexation behavior of scutellarein (SCUE) with 2-hydroxypropyl-ß-cyclodextrin (HP-ß-CD) has been investigated in both solution and in the solid state. SCUE/HP-ß-CD solid system was prepared by suspension method. The formation of SCUE/HP-ß-CD complex in aqueous solution was demonstrated by fluorescence spectroscopy, and the Job plot showed a maximum at a molar fraction of 0.5, indicating 1:1 inclusion complexation between SCUE and HP-ß-CD. However, SCUE/HP-ß-CD inclusion complex was characterized by means of XRD, DSC, (1)H, and two-dimensional NMR. Through the complexation between HP-ß-CD and SCUE, the water solubility and antitumor activity of SCUE were obviously increased. This satisfactory water solubility and high antitumor activity of the SCUE/HP-ß-CD complex will be potentially useful for its application on human colon cancer chemotherapies.

A novel polyrotaxane-based delivery system for scutellarin: preparation, characterization, and in vitro evaluation.

The safe and effective polyrotaxane-based drug delivery system could potentially increase the antiproliferative activity of antitumor medicine. A novel scutellarin-polyrotaxane (SCU-PR), in which scutellarin (SCU) was covalently bound to one of the hydroxyl groups of polyrotaxane (PR), was synthesized, and its characterization was further investigated by NMR, XRD, TG, DSC. The cytotoxicity of SCU-PR was assessed in vitro using human HCT116 and LOVO cell lines in results that the IC50 values of SCU-PR (1.03×10(-6) and 1.01×10(-6)mol/L, respectively), which compared with those of free SCU (7.80×10(-5) and 7.70×10(-5)mol/L, respectively), were lower. The valuable properties of SCU-PR will be potentially useful for its application on human colon cancer chemotherapies.