The placenta may play a key role in the activation of inflammation and initiation of insulin resistance (IR) during gestational diabetes mellitus (GDM) pathogenesis. Interleukin (IL)‑1β and IL‑18, regulated by NLR family pyrin domain containing‑3 (NLRP3) inflammasome, are important inflammatory cytokines in the initiation of maternal IR during GDM. However, the mechanism responsible for the regulatory of NLRP3 inflammasome in placenta remains unknown. Hydrogen sulfide (H₂S) exerts anti‑inflammatory function partially via suppressing the activation of the NLPR3 inflammasome. The present study aimed to investigate the role of NLRP3 inflammasome, H₂S synthetase cystathionine‑γ‑lyase (CSE) and cystathionine‑β‑synthetase (CBS) in placenta in the pathogenesis of GDM. Clinical placenta samples were collected from pregnant women with GDM (n=16) and healthy pregnant women at term (n=16). Western blot analysis was performed to detect the protein expression levels of NLRP3, cleaved caspase‑1, CBS and CSE in the placenta samples. Pearson's correlation analysis was performed to assess the correlation between NLRP3 inflammasome and H₂S synthetase. Human placental cells were cultured and treated with different concentrations of NaHS (0, 10, 25 and 50 nmol/l) or L‑cysteine (0, 0.25, 0.50 and 1.00 mmol/l). In addition, western blot analysis was performed to detect the protein expression levels of NLRP3 and cleaved caspase‑1, while ELISA was performed to measure the production of IL‑1β and IL‑18 in the culture media. The results demonstrated that the expression levels of NLRP3 and cleaved caspase‑1 increased, while the expression levels of CBS and CSE decreased in the placenta samples. In addition, the expression levels of NLRP3 and cleaved caspase‑1 were inversely correlated with the expression levels of CBS and CSE. Notably, NaHS and L‑cysteine significantly suppressed the expression levels of NLRP3 and cleaved caspase‑1, and the production of IL‑1 and IL‑18 in human placental cells. Taken together, the results of the present study suggest that H₂S synthetase deficiency in placenta may contribute to excessive activation of NLRP3 inflammasome in GDM.

Background. Irisin, an exercise-induced myokine and adipocytokine, has been reported to decrease in type 2 diabetic patients. Recently, several research studies indicated that circulating levels were correlated with bone mineral density (BMD). To evaluate bone metabolism, bone turnover markers (BTMs) should be included. However, with respect to newly diagnosed T2DM patients, the relevance of their irisin levels to their BTMs and BMD remains unclear. The investigation of serum irisin levels in patients who have been newly diagnosed with type 2 diabetes and illumination of the relationship between serum irisin levels and those two indices of BMD and BTMs mentioned above are the intention of this cross-sectional study. Methods. 66 new-onset type 2 diabetic patients (T2DM group), together with 82 control subjects (NGT group), were recruited in this study. Serum irisin concentrations and BTMs (including osteocalcin (OC), procollagen type 1 N-terminal propeptide (P1NP), and β-C-terminal telopeptides of type I collagen (β-CTX)) were determined by the enzyme-linked immunosorbent assay (ELISA). Glucose, lipid profile, and insulin were considered as measuring indicators as well. Dual-energy X-ray absorptiometry (DXA) was utilized to evaluate the indicator of BMD. Serum irisin, BTMs, and BMD were compared between diabetic patients and healthy individuals. Pearson and Spearman correlation analyses were applied as well to assess correlations between irisin and BTMs and BMD. Multiple stepwise regression analysis was conducted to identify the independent factors of irisin. ROC curve analyses were carried out for serum irisin prediction for osteoporosis/osteopenia (OP). Results. The serum levels of irisin, procollagen type 1, intact N-terminal propeptide (P1NP), and osteocalcin (OC) were evidently lower in T2DM subjects than in NGT subjects (10.90 ± 1.88 vs .11.69 ± 2.06 ng/mL, < 0.05; 36.42(25.68,51.70) vs. 44.52(35.73,58.05)ng/ml, < 0.05; 16.15(12.40,21.66) vs. 18.70(15.56, 23.22)ng/ml, < 0.05). Among patients with T2DM, the circulating irisin level of those with OP was lower than that of normal BMD (9.98 ± 2.09 vs. 11.39 ± 1.57 ng/ml, < 0.01); irisin had a negative correlation with β-C-terminal telopeptides of type I collagen (β-CTX) (r = −0.496, < 0.001) and came back unrelated to Lumbar BMD; Lumbar BMD was negatively relevant to OC (r = −0.274, < 0.05) and β-CTX (r = −0.410, < 0.01). Multiple linear regression analyses of stepwise models implied that TG, LDL-C, and β-CTX were independently associated with serum irisin concentrations ( < 0.01 or < 0.05). Conclusion. Serum irisin level was declined in patients with type 2 diabetes diagnosed in the near term and had a certain association with bone turnover markers. It is suggested to consider irisin as a potential biomarker of bone metabolic disorder in T2DM patients with the initial diagnosis.

Hydrogen sulfide (H₂S) has recently been identified as an endogenous gaseous signaling molecule. The aim of the present study was to investigate the effect of H₂S on high glucose- (HG-) induced ADAM17 expression and sFlt-1 production in 3T3-L1 adipocytes. Firstly, we found that HG DMEM upregulated the expression of ADAM17 and production of sFlt-1 in 3T3-L1 adipocytes. Knocking down ADAM17 attenuated the effect of high glucose on sFlt-1 production in adipocytes. HG decreased the expression of CSE and 3-MST, as well as the endogenous H₂S production. Furthermore, knocking down CSE and 3-MST significantly increased ADAM17 expression and sFlt-1 production. The addition of exogenous H₂S through the administration of sodium hydrosulfide (NaHS) inhibited HG-induced upregulation of ADAM17 expression and sFlt-1 production. In conclusion, decreased expression of CSE and 3-MST and the subsequent decrease in H₂S production contribute to high glucose-induced sFlt-1 production via activating ADAM17 in adipocytes. Exogenous H₂S donor NaHS has a potential therapeutic value for diabetic vascular complications.

Imbalance release of proangiogenic and antiangiogenic factor from placenta causes maternal endothelial dysfunction which is main manifestation of preeclampsia(PE). Hydrogen sulfide (H ₂ S) “the third endogenous gaseous signaling transmitter”, has been shown to have a pro‐angiogenic effect. We investigated whether H ₂ S modulate the release of proangiogenic and antiangiogenic factors in human placentas. It was found that H ₂ S synthesis enzymes cystathionine β‐synthase(CBS) and cystathionine γ‐lyase(CSE) were expressed in human placenta. The levels of CBS and CSE positively correlated with the levels of agiogenic factor vascular endothelial growth factor(VEGF) and inversely correlated to antiangiogenic factor soluble fms‐like tyrosine kinase 1(sFlt‐1) in placentas. H ₂ S precursor L‐cysteine significantly increased VEGFA release whereas decreased sFlt‐1 release in cultured placental explants andd placental trophoblasts. The effects of H ₂ S precursor could be reversed by siRNA of CBS and CSE. Moreover, H ₂ S decreased miR‐200c which target VEGF and increased miR‐133b which target sFlt‐1. The level of miR‐200c was significantly increased whereas miR‐133b decreased in PE placentas. Our results suggest that H ₂ S is an important factor that maintains the balance of proangiogenic factor VEGFA and antiangiogenic factor sFlt‐1 in placenta. Reduced H ₂ S generation in placentas leads to imbalance release of proangiogenic and antiangiogenic factor from placenta. Grant Funding Source : Natural Science Fundation of China No.81370734

Jinkui Shenqi Wan (JKSQW) is a classical traditional Chinese medicine (TCM) formula used for diabetic kidney disease (DKD), but its molecular mechanisms remain incompletely defined. While non-coding RNAs (ncRNAs) are increasingly recognized as key regulators in DKD, their role in the therapeutic action of JKSQW has not been systematically explored. Accordingly, we aimed to elucidate the potential mechanisms of JKSQW in DKD by integrating a circRNA–miRNA–mRNA regulatory network into a network pharmacology framework. Bioactive constituents and putative targets of JKSQW were retrieved from TCMSP and STITCH and mapped to UniProt. DKD-related differentially expressed mRNAs (DEmRNAs) were obtained from GEO (GSE1009; GPL16791) and analyzed using DESeq2 (Benjamini–Hochberg adjusted P < .05; |log2FC| > 2). circRNA–miRNA interactions were predicted using miRcode and cross-checked with starBase when available; miRNA–mRNA interactions were integrated from miRDB, miRTarBase, and TargetScan, retaining pairs supported by ≥ 2 resources or validated entries. Overlapping drug–disease targets underwent STRING-based protein–protein interaction (PPI) analysis, GO/KEGG enrichment with clusterProfiler, and molecular docking (AutoDock Vina). Docked complexes were visualized in PyMOL and annotated with PLIP. Fifty constituents mapped to 218 targets; 543 DEmRNAs were identified in DKD, with 20 overlapping genes considered potential therapeutic targets. PPI analysis highlighted hubs HTR2A, EGFR, TOP2A, CDK1, and MYC. Integration of ncRNA predictions yielded a circRNA–miRNA–mRNA network comprising 50 circRNAs, 32 miRNAs, and 20 mRNAs, revealing axes such as circ_0000962–miR‑15a‑5p–CDK1. Enrichment implicated p53 and AGE–RAGE signaling in apoptosis, fibrosis, and oxidative stress. Docking showed stigmasterol exhibited favorable affinities with HTR2A (−8.3 kcal/mol), EGFR (−7.6), TOP2A (−7.8), and CDK1 (−8.7). JKSQW may exert multi-component, multi-target protection against DKD by engaging ncRNA-mediated regulation and protein hubs within p53 and AGE–RAGE pathways. These findings provide a modern pharmacological rationale for this TCM formula and a transferable framework for ceRNA-integrated network analyses.