4,8-Dicarboxyl-8,9-iridoid-1-glycoside inhibits apoptosis in human osteoarthritis chondrocytes via enhanced c-MYC-mediated cholesterol metabolism in vitro.

BACKGROUND: Osteoarthritis (OA) is a degenerative disease related to cholesterol metabolism disorders. However, current therapies for OA are insufficient and no convincing disease-modifying OA drugs exist. Therefore, we aimed to elucidate the mechanism by which borojoa iridoid glycoside (BIG) inhibits chondrocyte apoptosis in OA. METHODS: Borojoa pulp was heated to 70 °C, and the main active substance in borojoa, BIG, was extracted by fractionation at an ultraviolet 254-nm absorption peak. Chondrocytes were identified by immunohistochemistry and visualized by immunofluorescence confocal microscopy. The proliferation of chondrocytes cultured with BIG was determined by MTS assay. The apoptosis of chondrocytes cultured with BIG was tested by Annexin V-FITC/PI, and the cytokine, protein, and cholesterol levels in chondrocytes were detected by ELISA, RT‒qPCR, Western blot, and biochemistry analyses. Protein‒protein interactions were verified by a coimmunoprecipitation (Co-IP) assay. RESULTS: BIG promoted chondrocyte proliferation and reduced apoptosis in vitro. BIG induced an alteration of the total RNA profiles in chondrocytes, and bioinformatic analysis showed that BIG inhibited chondrocyte apoptosis by promoting c-MYC expression; KEGG analysis confirmed that BIG-inhibited apoptosis was enriched in the cell cycle pathway. Flow cell cycle experiments confirmed that BIG promoted chondrocyte proliferation by significantly increasing the S phase cell number. The c-MYC inhibitor 10058-F4 stimulated the increased expression of IL-1ß, IL-6, TNF-α, and AGEs and suppressed the cholesterol metabolism, which promoted chondrocyte apoptosis and autophagy. Co-IP analysis showed that BIG promoted the interaction of c-MYC and CH25H, Bcl-2, which suggests that BIG could inhibit chondrocyte apoptosis in part by enhancing c-MYC-mediated cholesterol metabolism. CONCLUSIONS: This study confirmed that BIG promotes chondrocyte proliferation and inhibits apoptosis and autophagy, and BIG improving OA is associated with cholesterol metabolism. The results identify a potential mechanism by which BIG enhances c-MYC-mediated CH25H regulation of cholesterol metabolism in vitro and suggest that BIG might be a promising new drug against OA.

Association between neutrophil-to-lymphocyte ratio and postoperative fatigue in elderly patients with hip fracture.

Background: and purpose: Postoperative fatigue (POF) is a common and distressing post-operative symptom. This study aimed to explore the relationship between neutrophil-to-lymphocyte ratio (NLR) and POF in elderly patients with hip fracture. Method: Elderly patients (age ≥65 years) with acute hip fracture admitted to the Department of Orthopedics of Anqing Municipal Hospital from June 2018 to June 2020 were included. Fatigue was assessed using the Fatigue Severity Scale at the 3-month follow-up postoperatively. Univariate and multivariate analyses were performed to explore the associations between NLR and POF. The diagnostic performance of NLR was analysed using Receiver Operating Characteristic (ROC) curve analysis and the Delong test. Result: A total of 321 elderly patients with hip fractures were included; 120 (37.4 %) of them were diagnosed with POF. Univariate analysis indicated significant differences in NLR, platelet-to-lymphocyte ratio (PLR), education, neutrophil count, lymphocyte count, Hamilton Depression Scale (HAMD) and Insomnia Severity Index (ISI) scores (P < 0.05). Multivariate analysis indicated neutrophil count (odds ratio [OR], 1.46; 95 % confidence interval [CI] 1.27-1.67), lymphocyte count (OR 0.32, 95 % CI 0.19-0.53), NLR (OR1.81, 95 % CI 1.50-2.17) and PLR (OR 1.005, 95 % CI 1.001-1.009) were significantly associated with POF. The areas under the ROC curves (AUCs) of neutrophil count, lymphocyte count, NLR and PLR were 0.712, 0.667, 0.775 and 0.605, respectively. The Delong test indicated that NLR had the best diagnostic performance (p < 0.05). Conclusion: NLR independently predicts POF in elderly patients with acute hip fracture.

4,8-dicarboxyl-8,9-iridoid-1-glycoside Promotes Neural Stem Cell Differentiation Through MeCP2.

Background: Borojó (Borojoa patinoi Cuatrec) fruit has recently been shown to have a variety of health benefit, but the mechanisms have been little studied. The aim of this study was to investigate the effect of 4,8-dicarboxyl-8,9-iridoid-1-glycoside (388) on proliferation and differentiation of embryonic neural stem cells (NSCs). Methods: NSCs were treated with 388 and stem cell differentiation was determined by western blotting and immunofluorescence staining. The role of MeCP2 in 388-mediated embryonic NSCs differentiation was examined. Results: The results showed that in the presence of mitogen when NSCs proliferated and maintained their multipotency, treatment with 388 did not affect the viability of NSCs. Following mitogen withdrawal to initiate NSC differentiation, treatment with 388 at the doses of 10 and 50 µg/mL significantly increased neural differentiation in both cortex and spinal cord-derived culture. 388 also significantly up-regulated MeCP2 expression. The expression of the neuronal and oligodendrocytic markers was enhanced after addition of 388 in the differentiation culture. However, knockdown of MeCP2 results in inhibition of NSC differentiation, and the pro-differentiation effect of 388 was mostly abolished. Conclusions: This study confirmed that 388 stimulates differentiation of NSCs and identifies its mechanism of action by upregulating MeCP2.