Employing 16S rRNA sequencing and metabolomics analysis, the presence of the gut microbiota and its metabolites was determined. By means of immunofluorescence analysis, western blotting, and real-time PCR, the parameters of fatty acid metabolism, macrophage polarization, and the FFAR1/FFAR4-AMPK-PPAR pathway were subjected to detailed analysis. Using a LPS-stimulated RAW2647 cell model, the impact of FFAR1 and FFAR4 agonists on macrophage polarization was subsequently assessed.
The results of the study indicated that FMT, similar in effect to HQD, lessened UC by improving weight loss, restoring colon length, and reducing scores on both DAI and histopathological evaluations. Equally important, both HQD and FMT augmented the richness of the gut microbiota, influencing the composition of intestinal bacteria and their metabolites to create a new balance. Untargeted metabolomics demonstrated a substantial presence of fatty acids, especially long-chain fatty acids (LCFAs), in the HQD treatment, mitigating the DSS-induced ulcerative colitis (UC) through alteration of the gut microbial community. Consequently, FMT and HQD caused the recovery of fatty acid metabolism enzyme expression and simultaneously activated the FFAR1/FFAR4-AMPK-PPAR pathway, thus suppressing the NF-κB pathway. Cell-culture experiments revealed that the combination of HQD and FMT promoted macrophage polarization, specifically from an M1 to an M2 state, closely aligned with elevated anti-inflammatory cytokines and activated FFAR4.
Fatty acid metabolism modulation by HQD in ulcerative colitis (UC) is linked to the FFAR4-AMPK-PPAR pathway activation, resulting in M2 macrophage polarization.
A mechanism by which HQD combats UC is through its influence on fatty acid metabolism, ultimately promoting M2 macrophage polarization via the activation of the FFAR4-AMPK-PPAR pathway.
P. (Psoralea corylifolia L.) seeds For the treatment of osteoporosis in China, the plant corylifolia, popularly referred to as Buguzhi in traditional Chinese medicine, is often employed. Despite its identification as the key anti-osteoporosis constituent in P. corylifolia, psoralen (Pso) displays an unknown mechanism of action, along with unidentified molecular targets.
This study investigated the interplay between Pso and 17-hydroxysteroid dehydrogenase type 2 (HSD17B2), a protein involved in estrogen synthesis and suppressing the conversion of estradiol (E2) to address osteoporosis.
In mice, oral administration of an alkynyl-modified Pso probe (aPso) enabled in-gel imaging to analyze the tissue distribution of Pso. biotic fraction Using chemical proteomics, the liver's Pso target was identified and analyzed. The key action targets were confirmed by employing both co-localization methods and cellular thermal shift assays (CETSA). To characterize the key pharmacophore in Pso, the binding of Pso and its structural counterparts to HSD17B2 was evaluated through CETSA, HSD17B2 activity assessments, and in-gel visualization. Virtual docking simulations, alongside competitive testing of Pso with HSD17B2, in addition to scrutinizing the altered activity of mutated HSD17B2 and CETSA analysis, led to the localization of Pso's binding site within HSD17B2. Through ovariectomy-induced osteoporosis in mice, the in vivo effectiveness of Pso was determined. This was confirmed using micro-CT, histological H&E staining, measurements of HSD17B2 activity, and bone-specific biochemical assays.
The -unsaturated ester within Pso plays a crucial role as the pharmacophore, enabling Pso to regulate estrogen metabolism through its interaction with HSD17B2 within the liver. Irreversibly attaching to Lys236 of HSD17B2, Pso significantly reduces the activity of HSD17B2, preventing NAD's participation.
A journey into the binding pocket is not recommended. In vivo studies of ovariectomized mice found that Pso could suppress HSD17B2 enzyme activity, prevent the breakdown of E2, boost endogenous estrogen production, enhance bone metabolism markers, and possibly contribute to an anti-osteoporosis effect.
Hepatocyte HSD17B2's Lys236 residue is covalently targeted by Pso, effectively preventing E2 inactivation and potentially aiding in the treatment of osteoporosis.
Covalent binding of Pso to Lys236 of HSD17B2 in hepatocytes disrupts E2 inactivation, which may be therapeutically relevant for osteoporosis.
Tiger bone, a substance frequently utilized in traditional Chinese medicine, was believed to possess properties of wind-dispelling, pain-relieving, and strengthening sinews and bones, and was often applied in clinical contexts to treat bone blockages and bone atrophy. Jintiange (JTG), an artificial tiger bone substitute for natural tiger bone, has been approved by China's State Food and Drug Administration to mitigate osteoporosis symptoms, encompassing lumbago and backache, lassitude in the lower back and legs, flaccidity and weakness in the legs, and difficulty ambulating, based on Traditional Chinese Medicine (TCM). Biofuel production Natural tiger bone and JTG display comparable chemical compositions, characterized by the presence of minerals, peptides, and proteins. The compound's protective effect on bone loss in ovariectomized mice, along with its impact on osteoblast and osteoclast activity, has been documented. The detailed pathways by which peptides and proteins of JTG affect bone development are not completely clear.
Exploring the stimulating action of JTG proteins in the context of bone formation, with a focus on elucidating the associated underlying mechanisms.
JTG Capsules were processed using a SEP-PaktC18 desalting column to remove calcium, phosphorus, and other inorganic elements, ultimately isolating the JTG proteins. Investigations into the effects and underlying mechanisms of JTG proteins were conducted on MC3T3-E1 cells. Proliferation of osteoblasts was determined by employing the CCK-8 method. ALP activity was found using a relevant assay kit, and the bone mineralized nodules were stained by the alizarin red-Tris-HCl solution. Cell apoptosis analysis was performed using flow cytometry. Autophagy, as determined by MDC staining, was accompanied by the presence of autophagosomes, as seen under TEM. Laser confocal microscopy, employing immunofluorescence techniques, demonstrated nuclear localization of LC3 and CHOP. An examination of the expression levels of key proteins associated with osteogenesis, apoptosis, autophagy, PI3K/AKT and ER stress pathways was carried out through Western blot analysis.
Improved osteogenesis, a consequence of JTG protein action, was observed through modulation of MC3T3-E1 osteoblast proliferation, differentiation, mineralization, and the prevention of apoptosis, along with the promotion of autophagosome formation and autophagy. Their regulation also encompassed the expression of key proteins participating in the PI3K/AKT and ER stress pathways. By inhibiting PI3K/AKT and ER stress pathways, the regulatory effects of JTG proteins on osteogenesis, apoptosis, autophagy, and the PI3K/AKT and ER stress pathways can potentially be reversed.
JTG proteins' mechanism of promoting osteogenesis and inhibiting osteoblast apoptosis involves increasing autophagy, specifically through the PI3K/AKT and ER stress signaling cascade.
JTG proteins increased osteogenesis and decreased osteoblast apoptosis by bolstering autophagy, mediated by PI3K/AKT and endoplasmic reticulum stress signaling.
Radiotherapy can induce irradiation-related intestinal injury (RIII), often resulting in symptoms such as abdominal pain, diarrhea, nausea, vomiting, and even fatal outcomes. Wall's meticulous documentation of the Engelhardia roxburghiana. Leaves, a traditional Chinese herb, boasts a unique spectrum of anti-inflammatory, anti-tumor, antioxidant, and analgesic actions, employed in the treatment of damp-heat diarrhea, hernia, and abdominal pain, potentially offering protection from RIII.
To determine the protective influence of the full spectrum of flavonoids present in Engelhardia roxburghiana Wall. is the aim of this exploration. Leaves (TFERL) from RIII feature in the utilization of Engelhardia roxburghiana Wall.; furnish supporting literature. Leaves occupy a space in the extensive field of radiation protection.
Ionizing radiation (IR), administered at a lethal dose of 72Gy, enabled the observation of TFERL's impact on the survival of mice. To evaluate the protective effects of TFERL against RIII, a mouse model of RIII was created using 13 Gy of irradiation (IR). Haematoxylin and eosin (H&E) staining, along with immunohistochemistry (IHC), revealed the presence of small intestinal crypts, villi, intestinal stem cells (ISC), and ISC proliferation. To gauge the expression of genes relevant to intestinal integrity, quantitative real-time PCR (qRT-PCR) was utilized. Mice serum was scrutinized for the presence of superoxide dismutase (SOD), reduced glutathione (GSH), interleukin-6 (IL-6), and tumor necrosis factor- (TNF-). Laboratory-based cell models of RIII, exposed to irradiation levels of 2, 4, 6, and 8 Gray, were created. Using a clone formation assay, the radiation protective effect of TFERL on HIEC-6 cells, pre-treated with TFERL/Vehicle, was examined. Selleck Glafenine DNA damage was revealed by employing the comet assay and the immunofluorescence assay. Flow cytometry was used to detect the levels of reactive oxygen species (ROS), the cell cycle progression, and the rate of apoptosis. Proteins of interest, namely those related to oxidative stress, apoptosis, and ferroptosis, were detected by western blot analysis. The colony formation assay served to evaluate the impact of TFERL on the radiosensitivity of colorectal cancer cells, concluding the study.
Mice receiving TFERL treatment demonstrated improved survival and extended lifespan following a lethal radiation dose. Following irradiation-induced RIII in mice, TFERL mitigated RIII by reducing intestinal crypt/villi structural damage, augmenting the number and proliferation of intestinal stem cells, and upholding the integrity of the intestinal epithelial lining after total abdominal irradiation. Concurrently, TFERL facilitated the rise of irradiated HIEC-6 cells, along with a decrease in radiation-induced apoptosis and DNA damage. TFERL's role in promoting the expression of NRF2 and its cascade of antioxidant proteins has been meticulously explored through mechanistic studies. Importantly, the suppression of NRF2 activity was directly linked to the loss of TFERL's radioprotective abilities, firmly establishing the NRF2 pathway as critical to TFERL's radiation-protective function.