Auricularia auricula Anionic Polysaccharide Nanoparticles for Gastrointestinal Delivery of Pinus koraiensis Polyphenol Used in Bone Protection under Weightlessness.

Auricularia auricula polysaccharides used in Pinus koraiensis polyphenol encapsulation and delivery under weightlessness are rarely reported. In this study, an anionic polysaccharide fragment named AAP Iα with a molecular weight of 133.304 kDa was isolated and purified to construct a polyphenol encapsulation system. Nanoparticles named NPs-PP loaded with a rough surface for Pinus koraiensis polyphenol (PP) delivery were fabricated by AAP Iα and ε-poly-L-lysine (ε-PL). SEM and the DLS tracking method were used to observe continuous changes in AAP Iα, ε-PL and PP on the nanoparticles' rough surface assembly, as well as the dispersion and stability. Hydrophilic, monodisperse and highly negative charged nanoparticles can be formed at AAP Iα 0.8 mg/mL, ε-PL 20 µg/mL and PP 80 µg/mL. FT-IR was used to determine their electrostatic interactions. Release kinetic studies showed that nanoparticles had an ideal gastrointestinal delivery effect. NPs-PP loaded were assembled through electrostatic interactions between polyelectrolytes after hydrogen bonding formation in PP-AAP Iα and PP-ε-PL, respectively. Colon adhesion properties and PP delivery in vivo of nanoparticles showed that NPs-PP loaded had high adhesion efficiency to the colonic mucosa under simulated microgravity and could enhance PP bioavailability. These results suggest that AAP Iα can be used in PP encapsulation and delivery under microgravity in astronaut food additives.

Silencing of miR-483-5p alleviates postmenopausal osteoporosis by targeting SATB2 and PI3K/AKT pathway.

Postmenopausal osteoporosis (PMOP) poses a significant threat to women's health worldwide. However, detailed molecular mechanism and therapeutic strategy for PMOP remain insufficient. Accumulating evidence suggests that miR-48-5p is implicated in the pathogenesis of osteoporosis. The present study aimed to determine the role and mechanism of miR-483-5p in PMOP. Results from PMOP patients demonstrated that miR-483-5p was up-regulated and SATB2 was down-regulated. Luciferase reporter assay identified SATB2 as a direct target gene of miR-483-5p. Experiments in MC3T3-E1 cells indicated that miR-483-5p mimic markedly inhibited cell viability as well as the expressions of OPG, RUNX2 and BMP2. And miR-483-5p inhibitor, SATB2-overexpressed lentiviruses (Lv-SATB2) or LY294002 (PI3K/AKT inhibitor) significantly reversed the above results. Similarly, PI3K/AKT signaling was activated by miR-483-5p mimic, and was inhibited in miR-483-5p inhibitor, Lv-SATB2 or LY294002 treated cells. In vivo experiments showed that miR-483-5p inhibitor significantly increased the bone mineral density and biomechanical parameters of femurs in ovariectomized (OVX) rats by targeting SATB2. In addition, the osteogenic differentiation and PI3K/AKT signaling were also regulated by miR-483-5p-SATB2 axis. Taken together, our findings indicated that miR-483-5p contributed to the pathogenesis of PMOP by inhibiting SATB2 and activating PI3K/AKT pathway. MiR-483-5p/SATB2 could be selected as a potential therapeutic target for PMOP.

MiR-151a-3p Promotes Postmenopausal Osteoporosis by Targeting SOCS5 and Activating JAK2/STAT3 Signaling.

Postmenopausal osteoporosis, the most common type of primary osteoporosis, poses a significant threat to women's health worldwide. However, detailed molecular mechanism and therapeutic strategy for postmenopausal osteoporosis remain insufficient. Increasing evidence suggests that microRNAs contributed to the pathogenesis of osteoporosis and could be considered as potential therapeutic targets. In this study, we found that miR-151a-3p was upregulated in osteoporosis samples. Experiments in MC3T3-E1 cells indicated that miR-151a-3p significantly inhibited cell viability and promoted lactate dehydrogenase release, as well as increased RANKL/OPG ratio and decreased Runx2 and BMP2 expressions. SOCS5 was identified as a direct target gene of miR-151a-3p, which was confirmed by luciferase reporter assay. Moreover, an inverse correlation between miR-151a-3p and SOCS5 was observed in osteoporosis femurs. In addition, JAK2/STAT3 pathway was found to be involved in the progress of osteoporosis mediated by miR-151a-3p-SOCS5 axis. In vivo, ovariectomized (OVX) rat model was established to simulate postmenopausal osteoporosis. The results revealed that miR-151a-3p significantly decreased the bone mineral density and biomechanical parameters of femurs in OVX rats by targeting SOCS5, and that JAK2/STAT3 pathway is a downstream target of miR-151a-3p-SOCS5 axis in OVX rats. In conclusion, our findings suggested that miR-151a-3p contributed to the pathogenesis of postmenopausal osteoporosis, and promoted its progress by targeting SOCS5 and activating JAK2/STAT3 signaling. Thus, anti-miR-151a-3p could be a potential therapeutic strategy for postmenopausal osteoporosis.

Efficacy of Xixiancao (Herba Siegesbeckiae Orientalis) on interactions between nuclear factor kappa-B and inflammatory cytokines in inflammatory reactions of rat synovial cells induced by sodium urate.

OBJECTIVE: To investigate the interaction between nuclear factor kappa-B (NF-κB) and inflammatory cytokines in synovial cell inflammatory responses induced by sodium urate, and to evaluate the efficacy of Xixiancao (Herba Siegesbeckiae Orientalis) on these interactions. METHODS: The interactions between NF-κB and inflammatory cytokines/mediators in synovial cells in acute gouty arthritis were investigated. We observed the expressions of NF-κB, interleukin (IL)-1ß, IL-8, and tumor necrosis factor alpha (TNF-α) in synovial cells at different timepoints in an in vitro model of synovial cell inflammatory responses induced by sodium urate and in an in vivo model of gouty arthritis. Changes in the expressions of NF-κB, IL-1ß, IL-8, and TNF- in synovial cells of all experimental groups were compared and observed after treatment with different doses of Xixiancao (Herba Siegesbeckiae Orientalis) and colchicine. The interactions between NF-κB and IL-1ß, IL-8, and TNF-α were analyzed. Pathological changes in synovial tissues were observed in rats with acute gouty arthritis. RESULTS: Compared with the blank group, the expression levels of NF-κB, IL-1ß, IL-8, and TNF-α were increased significantly at different timepoints in the in vitro model of synovial cell inflammatory responses induced by sodium urate, and in the in vivo model of gouty arthritis. Compared with the model group, the expressions of NF-κB, IL-1ß, IL-8, and TNF-α in synovial cells induced by sodium urate were decreased in the different Xixiancao (Herba Siegesbeckiae Orientalis) dose groups and the colchicine group. The effect was more obvious in the high dose Xixiancao (Herba Siegesbeckiae Orientalis) group. The expression of NF-κB in synovial cells was positively correlated with the expressions of IL-1ß, IL-8, and TNF-. Histopathological examination of synovial tissues in the high dose Xixiancao (Herba Siegesbeckiae Orientalis) group and Colchicine group showed that the characteristics of acute gouty arthritis were reduced, and there was a trend towards a positive correlation between NF-κB and inflammatory cytokine expressions. CONCLUSION: The activation of NF-κB is associated with the activation of IL-1ß, IL-8, and TNF-α during the pathogenesis of acute gouty arthritis, leading to the continuation and enhancement of the inflammatory response. Expressions of IL-1ß, IL-8, and TNF-α in synoviocytes during acute gouty arthritis effectively inhibit local inflammation.

High-Throughput Metabolomics Method for Discovering Metabolic Biomarkers and Pathways to Reveal Effects and Molecular Mechanism of Ethanol Extract From Epimedium Against Osteoporosis.

Metabolomics is an effective strategy to explore the molecular mechanism of herbal medicine. Epimedium, a traditional Chinese herb from the Epimedium brevicornu Maxim., has a therapeutic effect on osteoporosis (OP), however the molecular mechanism of the anti-OP effect is uncle\ar. Therefore, we investigated the pharmacological effect and action mechanism of ethanol extract of epimedium (Ext-epi) onOP rat model. The serum of OP rats was analyzed utilized UPLC-Q-TOF/MS metabolomics, and the potential biomarkers were screened and identified using multivariate data analysis systems and network databases. To further appraise the influence of Ext-epi on biological markers and metabolic pathways, and reveal the potential mechanism of Ext-epi on OP treatment. The results showed that 46 potential biomarkers were screened out and after intervention with Ext-epi extracts solution, 16 potential biomarkers were significantly recalled. Further pathway experiments showed that key pathway analysis include sarachidonic acid metabolism, glycerolphospholipid metabolism as potential targets which is related with the efficacy of Ext-epi protect against OP. These results explain the correlation between metabolites and molecular mechanisms, which is of great significance for understanding the intervention of Ext-epi on OP. In short, based on UPLC-Q-TOF/MS metabolomics may provide effective strategies for understanding the pathogenesis of diseases and evaluating the intervention effect of natural products.

Z-Guggulsterone attenuates glucocorticoid-induced osteoporosis through activation of Nrf2/HO-1 signaling.

AIMS: The present study aims to investigate the protective effect and underlying mechanism of Z-Guggulsterone (Z-GS), an active component from myrrh, on glucocorticoid-induced osteoporosis (GIO). MAIN METHODS: GIO rats were used to simulate osteoporosis in vivo while MC3T3-E1 cells were induced to osteoblast differentiation and treated with dexamethasone to simulate osteoporosis in vitro. The rats and cells were treated with Z-GS according to the protocol. The bone mineral density, biomechanical parameters and microstructure of GIO rats were measured with appropriate devices. Cell viability of MC3T3-E1 cells were analyzed via CCK-8 assay. Bone turnover markers and oxidative stress markers were detected by ELISA, and the expressions of Nrf2 and HO-1 were assessed by western blot. siRNA-Nrf2 and siRNA-HO-1 were transfected in MC3T3-E1 cells to knockdown the expressions of Nrf2 and HO-1. KEY FINDINGS: Z-GS significantly increased the body weights and bone mineral density, ameliorated the femoral biomechanical parameters and microstructure of GIO rats. Z-GS treatment also reversed DXM-induced changes of bone turnover markers and oxidative stress in rats and MC3T3-E1 cells. The expressions of Nrf2 and HO-1 were inhibited in the model group and treatment with Z-GS could markedly increase their expressions. Nrf2 or HO-1 knockdown observably abrogated the beneficial role of Z-GS on cells. SIGNIFICANCE: Our results demonstrated that Z-GS exerted bone protective and antioxidant stress properties through activation of Nrf2/HO-1 signaling in GIO models in vivo and in vitro. Therefore, Z-GS could be considered as a promising candidate for the treatment of GIO.

Combined low-dose LiCl and LY294002 for the treatment of osteoporosis in ovariectomized rats.

BACKGROUND: To provide a low-toxicity and high-efficacy clinical treatment for osteoporosis via a novel combination of LiCl and LY294002. METHODS: The protein levels of p-AKT, AKT, p-GSK3ß, GSK3ß, ß-catenin, p-ß-catenin, and NFATC1 were measured in osteoblasts and osteoclasts by Western blot. ALP activity and TRACP activity were measured using the corresponding kit. The levels of BALP, PINP, CTX, and TRACP-5b were determined in accordance with the requirements of the ELISA kits. Microstructural analysis was performed on the left distal femur using microcomputed tomography. RESULTS: Treatment with the combination of LiCl and LY294002 led to a markedly increased osteoblast activity but significantly decreased osteoclast differentiation and bone absorption capacity compared with the treatment with LiCl or LY294002 alone (P < 0.01). In serum, the low-dose combination of LiCl and LY294002 significantly enhanced BALP levels (P < 0.01) and significantly decreased PINP, TRACP-5b, and CTX levels (P < 0.01) compared with the application of either drug alone. CONCLUSIONS: This study indicates that drug combinations directed at multiple targets could be used for osteoporosis treatment by promoting osteoblast proliferation and inhibiting differentiation with high efficiency.

MicroRNA-148a-3p promotes survival and migration of endothelial cells isolated from Apoe deficient mice through restricting circular RNA 0003575.

The incidence of atherosclerosis is greatly increased, which becomes the leading cause for the death and disability worldwide. Endothelial cells dysfunction plays a substantial role in the pathogenesis of atherosclerosis. MicroRNA-148a-3p (miR-148a-3p) and circular RNA 0003575 (circ_0003575) modulated lipid metabolism and proliferative function of endothelial cells, respectively. However, the role of them in modulation of endothelial cell function and progression of atherosclerosis remains unknown. Endothelial cells were isolated from the aorta of Apoe-/- mice. miR-148a-3p in atherosclerosis patients and healthy controls were measured by qRT-PCR. Overexpression and knockdown of miR-148a-3p in endothelial cells were established. The proliferation, migration and apoptosis of endothelial cells were measured by MTT, Transwell, and fluorescence microscope, respectively. Online software (miRWalk 2.0 and RegRNA2.0) and databases (miRWalk, miRanda, RNA22, and Targetscan) were used to predict potential target genes of miR-148a-3p and circ_0003575. The expression of target genes was detected through western blotting. The expression of miR-148a-3p was significantly upregulated in patients with atherosclerosis as relative to healthy people. Overexpression of miR-148a-3p exhibited stimulatory effects on endothelial cell proliferation and migration and inhibited programmed cell death. Six intersection target genes, c-MAF, FOXO4, FOXO3, MITF, ETV7, and CRX, were predicted between miR-148a-3p and circ_0003575. The opposite effects of circ_0003575 and miR-148a-3p on the expression of FOXO4 and FOXO3, which are essential for lipid metabolism. We demonstrate that miR-148a-3p suppresses FOXO4 and FOXO3 expression via interruption of circ_0003575 function, which in turn impairs the proliferative and migratory function of endothelial cells, eventually exacerbating the atherosclerosis.

High-throughput metabolomics investigates anti-osteoporosis activity of oleanolic acid via regulating metabolic networks using ultra-performance liquid chromatography coupled with mass spectrometry.

BACKGROUND: Osteoporosis has brought about heavy socio-economic burden in the morbidity and medical expenses associated with osteoporosis treatment and various restrictions on behavior of their social roles. Oleanolic acid (OA) is an anti-osteoporosis natural product, but molecular mechanisms of therapeutic effect are not still well known. PURPOSE: In this study, we explore anti-osteoporosis activity of oleanolic acid and predict the underlying mechanisms by metabolomics strategy. METHODS: SD rats were intraperitoneal injection with prednison for once to establish osteoporosis model. Using metabolomics strategy based on ultra-performance liquid chromatography coupled with quadrupole time-of-flight/ mass spectrometry (UPLC-TOF/MS), serum samples of 24 rats were analyzed to seek differential metabolites and pathway associated with OA treatment of osteoporosis. In addition, the effect of OA on osteoporosis rats was also evaluated by clinical biochemistry indicators and bone density analysis. RESULTS: Clinical biochemistry indicators and bone density of lumbar and femur were reversed by OA treatment. A total of 25 potential biomarkers were identified in the rats model of glucocorticoid-induced osteoporosis, and oleanolic acid have a regulatory effect on 17 of them that related to some vital metabolic pathway such as linoleic acid metabolism, valine, leucine and isoleucine biosynthesis, phenylalanine, tyrosine and tryptophan biosynthesis as well as cysteine and methionine metabolism. The ingenuity pathway analysis (IPA) platform is applied to further understanding the relationship between metabolic changes and therapeutic effect of OA, which the disordered state carbohydrate metabolism, molecular transport and lipid metabolism in glucocorticoid-induced osteoporosis rats are mainly ameliorated by oleanolic acid. CONCLUSION: Metabolomics provides a novel method to investigate the anti-osteoporosis effects of OA and probe into the potential mechanisms, and will contributes to the development of new drugs.

Pine polyphenols from Pinus koraiensis prevent injuries induced by gamma radiation in mice.

Pine polyphenols (PPs) are bioactive dietary constituents that enhance health and help prevent diseases through antioxidants. Antioxidants reduce the level of oxidative damages caused by ionizing radiation (IR). The main purpose of this paper is to study the protective effect of PPs on peripheral blood, liver and spleen injuries in mice induced by IR. ICR (Institute of Cancer Research) male mice were administered orally with PPs (200 mg/kg b.wt.) once daily for 14 consecutive days prior to 7 Gy γ-radiations. PPs showed strong antioxidant activities. PPs significantly increased white blood cells, red blood cells and platelets counts. PPs also significantly reduced lipid peroxidation and increased the activities of superoxide dismutase, catalase and glutathione peroxidases, and the level of glutathione. PPs reduced the spleen morphologic injury. In addition, PPs inhibited mitochondria-dependent apoptosis pathways in splenocytes induced by IR. These results indicate that PPs are radioprotective promising reagents.