Novel deoxyhypusine synthase (DHPS) inhibitors target hypusination-induced vasculogenic mimicry (VM) against malignant melanoma.

Vasculogenic mimicry (VM) contributes factor to the poor prognosis of malignant melanoma. Developing deoxyhypusine synthase (DHPS) inhibitors against melanoma VM is clinically essential. In this study, we optimized and synthesized a series of compounds based on the candidate structure, and the hit compound 7k was identified through enzyme assay and cell viability inhibition screening. Both inside and outside the cell, 7k's ability to target DHPS and its high affinity were demonstrated. Molecular dynamics and point mutation indicated that mutations of K329 or V129 in DHPS abolish 7k's inhibitory activity. Using PCR arrays, solid-state antibody microarrays, and angiogenesis assays investigated 7k's impact on melanoma cells to reveal that DHPS regulates melanoma VM by promoting FGFR2 and c-KIT expression. Surprisingly, 7k was discovered to inhibit MC1R-mediated melanin synthesis in the zebrafish. Pharmacokinetic evaluations demonstrated 7k's favorable properties, and xenograft models evidenced its notable anti-melanoma efficacy, achieving a TGI of 73 %. These results highlighted DHPS as key in melanoma VM formation and confirmed 7k's potential as a novel anti-melanoma agent.

A systematic review and meta-analysis of acupuncture for De Quervain's tenosynovitis treatment.

BACKGROUND: De Quervain's tenosynovitis (DQt) is a prevalent chronic inflammatory musculoskeletal disorder predominantly affecting the radial aspect of the wrist. This study conducted a comprehensive review of the efficacy of acupuncture in treating De Quervain's tenosynovitis (DQt). Although there is evidence suggesting that acupuncture can alleviate symptoms of DQt-characterized by pain, swelling, and functional impairment-higher-level evidence is still required to further substantiate its efficacy and safety. This study conducted a comprehensive review of the efficacy of acupuncture in treating De Quervain's tenosynovitis (DQt). METHODS: By systematically searching databases such as PubMed, Science Direct, Web of Science, Google Scholar, EMbase, PEDro, China National Knowledge Infrastructure Database (CNKI), Wanfang Database, and Chongqing VIP China Science, Technology Journal Database (VIP), we retrieved randomized controlled trial (RCT) literature on acupuncture for DQt, with the search period extending to November 1, 2023. After extracting and assessing data from the included literature, we performed Meta-analysis using RevMan 5.4.1 software. RESULTS: The results encompassed 14 RCT papers, involving 851 patients. The Meta-analysis findings indicated that, when compared to topical analgesics, acupuncture demonstrated a significant increase in treatment effectiveness (RR = 1.24; 95% CI = 1.11, 1.39, P = 0.0002) and a notable reduction in VAS pain scores (MD = -1.06; 95% CI = -1.51, -0.61, P < 0.00001). However, no statistically significant difference was observed in conney wrist joint scores. Furthermore, acupuncture was found to reduce VAS pain scores compared to the waiting list group. In comparison to corticosteroid injections (CSI), acupuncture did not show statistical significance in VAS, effectiveness rate, and conney wrist scores. CONCLUSION: Acupuncture exhibited a promising trend in alleviating pain associated with DQt and enhancing treatment effectiveness. Nonetheless, due to limitations in the quantity and quality of the included studies, these findings warrant further validation through additional research.

An Investigation into the Interaction between Double Hydroxide-Based Antioxidant Benzophenone Derivatives and Cyclooxygenase 2.

Cyclooxygenases 2 (COX2) is a therapeutic target for many inflammation and oxidative stress associated diseases. A high-throughput technique, biolayer interferometry, was performed to primarily screen the potential COX2 binding activities of twelve newly synthesized double hydroxide-based benzophenone derivatives. Binding confirmation was achieved by molecular docking and multi-spectroscopy studies. Such a combined method provided a comprehensive understanding of binding mechanism and conformational changes. Compounds DB2, SC2 and YB2 showed effective COX2 binding activity and underlined the benefits of three phenolic hydroxyl groups adjacent to each other on the B ring. The twelve tested derivatives were further evaluated for antioxidant activity, wherein compound SC2 showed the highest activity. Its concentration for the 50% of maximal effect (EC50) value was approximately 1000 times greater than that of the positive controls. SC2 treatment effectively improved biochemical indicators caused by oxidative stress. Overall, compound SC2 could serve as a promising candidate for further development of a new potent COX2 inhibitor.

DHPS-Mediated Hypusination Regulates METTL3 Self-m6A-Methylation Modification to Promote Melanoma Proliferation and the Development of Novel Inhibitors.

Discovering new treatments for melanoma will benefit human health. The mechanism by which deoxyhypusine synthase (DHPS) promotes melanoma development remains elucidated. Multi-omics studies have revealed that DHPS regulates m6A modification and maintains mRNA stability in melanoma cells. Mechanistically, DHPS activates the hypusination of eukaryotic translation initiation factor 5A (eIF5A) to assist METTL3 localizing on its mRNA for m6A modification, then promoting METTL3 expression. Structure-based design, synthesis, and activity screening yielded the hit compound GL-1 as a DHPS inhibitor. Notably, GL-1 directly inhibits DHPS binding to eIF5A, whereas GC-7 cannot. Based on the clarification of the mode of action of GL-1 on DHPS, it is found that GL-1 can promote the accumulation of intracellular Cu2+ to induce apoptosis, and antibody microarray analysis shows that GL-1 inhibits the expression of several cytokines. GL-1 shows promising antitumor activity with good bioavailability in a xenograft tumor model. These findings clarify the molecular mechanisms by which DHPS regulates melanoma proliferation and demonstrate the potential of GL-1 for clinical melanoma therapy.

Pulsed electromagnetic field stimulates osteoprotegerin and reduces RANKL expression in ovariectomized rats.

Pulsed electromagnetic field (PEMF) has been shown to increase bone mineral density in osteoporosis patients and prevent bone loss in ovariectomized rats. But the mechanisms through which PEMF elicits these favorable biological responses are still not fully understood. Receptor activator of nuclear factor κB ligand (RANKL) and osteoprotegerin (OPG) are cytokines predominantly secreted by osteoblasts and play a central role in differentiation and functional activation of osteoclasts. The purpose of this study was to investigate the effects of PEMF on RANKL and OPG expression in ovariectomized rats. Thirty 3-month-old female Sprague-Dawley rats were randomly divided into three groups: sham-operated control (Sham), ovariectomy control (OVX), and ovariectomy with PEMF treatment (PEMF). After 12-week interventions, the results showed that PEMF increased serum 17ß-estradiol level, reduced serum tartrate-resistant acid phosphatase level, increased bone mineral density, and inhibited deterioration of bone microarchitecture and strength in OVX rats. Furthermore, PEMF could suppress RANKL expression and improve OPG expression in bone marrow cells of OVX rats. In conclusion, this study suggests that PEMF can prevent ovariectomy-induced bone loss through regulating the expression of RANKL and OPG.

Hypusination-induced DHPS/eIF5A pathway as a new therapeutic strategy for human diseases: A mechanistic review and structural classification of DHPS inhibitors.

The discovery of new therapeutic strategies for diseases is essential for drug research. Deoxyhypusine synthase (DHPS) is a critical enzyme that modifies the conversion of the eukaryotic translation initiation factor 5A (eIF5A) precursor into physiologically active eIF5A (eIF5A-Hyp). Recent studies have revealed that the hypusine modifying of DHPS on eIF5A has an essential regulatory role in human diseases. The hypusination-induced DHPS/eIF5A pathway has been shown to play an essential role in various cancers, and it could regulate immune-related diseases, glucose metabolism-related diseases, neurological-related diseases, and aging. In addition, DHPS has a more defined substrate and a well-defined structure within the active pocket than eIF5A. More and more researchers are focusing on the prospect of advanced development of DHPS inhibitors. This review summarizes the regulatory mechanisms of the hypusination-induced DHPS/eIF5A pathway in a variety of diseases in addition to the inhibitors related to this pathway; it highlights and analyzes the structural features and mechanisms of action of DHPS inhibitors and expands the prospects of future drug development using DHPS as an anticancer target.

SIRT1-dependent mitochondrial biogenesis supports therapeutic effects of 4-butyl-polyhydroxybenzophenone compounds against NAFLD.

The mitochondria have been identified as key targets in nonalcoholic fatty liver disease (NAFLD), one of the most prevalent chronic liver damage diseases globally. Meanwhile, the biological information analysis in this study revealed that SIRT1, PPARG, PPARA, and PPARGC1A (mitochondrial biogenesis-related proteins) were NAFLD therapeutic targets. Therefore, the design and synthesis of targeted drugs that promote mitochondrial biogenesis and improve mitochondrial function are particularly important for NAFLD treatment. Recently, we introduced butyls, hydroxyls, and halogens to benzophenone and synthesized a series of NAFLD-related 4-butylpolyhydroxybenzophenone compounds, aiming at investigating the hepatoprotective activity from the aspect of mitochondrial biogenesis. The structure-activity relationship demonstrated that hydroxyl and ketone groups were active groups interacting with mitochondrial biogenesis proteins (SIRT1 and PGC1α), and the activity was stronger when the o-hydroxyl group was present on the benzene ring. In contrast, the activity was little affected by the presence of the p-hydroxyl group, m-hydroxyl group, butyl group type, or halogen. In addition, in vitro studies confirmed that these compounds could directly bind to SIRT1 and PGC1α, markedly promote their interaction, significantly increase the expression of proteins and genes related to mitochondrial biogenesis (SIRT1, PGC1α, NRF1, TFAM, COX1, and ND6) and subsequently ameliorate mitochondria dysfunction, which was evidenced by the decreased ROS, upregulated ATP production, increased MMP, and enhanced mitochondrial number. According to the outcomes of our in vitro and in vivo experiments, 4-butyl-polyhydroxybenzophenone compounds could also effectively reduce the formation of lipid droplets and liver injury index (ALT, AST, LDH, AKP, γ-GT, and GDH) and improve the level of antioxidant enzymes (GSH and SOD). Particularly, the treatment of these compounds after a high-fat diet could significantly reduce body weight, decrease liver coefficient, attenuate liver damage, and ameliorate lipid accumulation in rat liver, demonstrating their therapeutic effects on NAFLD. Mechanistically, 4-butyl-polyhydroxybenzophenone compounds promoted mitochondrial biogenesis and eventually prevented NAFLD liver injury by activating the PGC1α signaling pathway in a SIRT1-dependent manner, which was strongly supported by SIRT1 inhibitor EX527.

Improving Thermostability and Catalytic Activity of Glycosyltransferase From Panax ginseng by Semi-Rational Design for Rebaudioside D Synthesis.

As a natural sweetener and sucrose substitute, the biosynthesis and application of steviol glycosides containing the component rebaudioside D have attracted worldwide attention. Here, a glycosyltransferase PgUGT from Panax ginseng was first reported for the biosynthesis of rebaudioside D. With the three-dimensional structures built by homology modeling and deep-learning-based modeling, PgUGT was semi-rationally designed by FireProt. After detecting 16 site-directed variants, eight of them were combined in a mutant Mut8 with both improved enzyme activity and thermostability. The enzyme activity of Mut8 was 3.2-fold higher than that of the wild type, with an increased optimum reaction temperature from 35 to 40°C. The activity of this mutant remained over 93% when incubated at 35°C for 2 h, which was 2.42 times higher than that of the wild type. Meanwhile, when the enzymes were incubated at 40°C, where the wild type was completely inactivated after 1 h, the residual activity of Mut8 retained 59.0% after 2 h. This study would provide a novel glycosyltransferase with great potential for the industrial production of rebaudioside D and other steviol glycosides.

Isolation and Quantification of the Hepatoprotective Flavonoids From Scleromitron diffusum (Willd.) R. J. Wang With Bio-Enzymatic Method Against NAFLD by UPLC-MS/MS.

Flavonoids were the major phytochemicals against hepatic peroxidative injury in Scleromitron diffusum (Willd.) R. J. Wang with an inventive bio-enzymatic method by our group (LU500041). Firstly, the total flavonoids from Scleromitron diffusum (Willd.) R. J. Wang were extracted by reflux, ultrasonic, ultrasound-assisted enzymatic methods (TFH), and the bio-enzymatic method (Ey-TFH). Then 24 flavonoid compounds were isolated and quantified in the extracts by UPLC-MS/MS. Next, six representative differential compounds in Ey-TFH were further screened out by multivariate statistical analysis compared with those in TFH. In a further step, Ey-TFH presented a higher protective rate (59.30 ± 0.81%) against H2O2-damaged HL-02 hepatocytes than TFH. And six representative differential compounds at 8 and 16 µmol/L all exerted significant hepatoprotective effects (p < 0.05 or p < 0.01). Finally, the therapeutic action of Ey-TFH for nonalcoholic fatty liver disease (NAFLD) was processed by a rat's model induced with a high-fat diet. Ey-TFH (90, 120 mg/kg) significantly ameliorated the lipid accumulation in the rat model (p < 0.05). Meanwhile, Ey-TFH relieved liver damage. The levels of ALT, ALP, AST, LDH, and γ-GT in rats' serum were also significantly reduced (p < 0.05 or p < 0.01). In addition to this, the body's antioxidant capacity was improved with elevated SOD and GSH levels (p < 0.05) and down-regulated MDA content (p < 0.01) after Ey-TFH administration. Histopathological observations of staining confirmed the hepatic-protective effect of Ey-TFH.

Effects of electro-acupuncture on oestrogen levels, body weight, articular cartilage histology and MMP-13 expression in ovariectomised rabbits.

BACKGROUND: Electro-acupuncture (EA) treatment has been shown to decrease pain and improve the function of ovariectomised (OVX) rats with osteoarthritis (OA); however, the underlying mechanisms remain unclear. OBJECT: We used OVX rabbits to replicate natural human menopausal processes and to evaluate whether EA could be used to prevent and treat postmenopausal OA. METHODS: The rabbits were randomly divided into four groups of eight: a normal control group (NC), an OVX group, an ERT group (oestrogen replacement therapy after OVX) and an EA group (EA therapy after OVX). After the interventions, all of the animals were killed. Serum oestrogen levels and body weight were measured. The transcription of matrix metalloproteinase-13 (MMP-13) mRNA was detected using reverse transcriptase-PCR. Modified Mankin scores were used for histological assessment. Expression of MMP-13 in cartilage was determined by immunohistochemistry. RESULTS: Both the EA group and the ERT group had increased serum oestrogen levels (p=0.028, p=0.037 respectively), as well as decreased expression of MMP-13 (p=0.000, p=0.000, respectively), relative to the OVX group. The body weight of the EA group was lower than that of the OVX group and the NC group (p=0.007), as well as the ERT group (p=0.010). CONCLUSIONS: EA could be a new method for preventing and treating postmenopausal OA by producing endogenous sex hormones that inhibit the expression of MMP-13 and cause weight loss with no side effects and a relatively low cost.

Electroacupuncture prevents ovariectomy-induced osteoporosis in rats: a randomised controlled trial.

BACKGROUND: Electroacupuncture (EA) treatment has been shown to increase bone mineral density (BMD) in ovariectomised (OVX) rats; however, the underlying mechanisms remain unclear. OBJECTIVE: To systematically evaluate the effects of EA on OVX rats and the Wnt/ß-catenin signalling pathway. METHODS: Three-month-old female Sprague-Dawley rats were randomly divided into three different groups (n=10 each): sham operated control (sham operated), ovariectomy (OVX) and ovariectomy with EA treatment (OVX+EA). Rats in the OVX+EA group received 12-week EA treatments. RESULTS: Serum bone-specific alkaline phosphatase level (p<0.01), BMD of the proximal femoral metaphysis and the fifth lumbar (L5) vertebral body (both, p<0.05) and maximum load and energy to failure of L5 vertebral body (both p<0.01) were significantly higher in the OVX+EA group than in the OVX group. Trabecular area, trabecular width and trabecular number were significantly higher in the OVX+EA group by 66.9%, 29.2% and 30.3%, respectively, than in the OVX group (all, p<0.01). Trabecular separation was 31.9% lower in the OVX+EA group than in the OVX group (p<0.01). Quantitative real-time reverse transcription polymerised chain reaction indicated that the expressions of mRNAs for low-density lipoprotein receptor-related protein 5 and ß-catenin were significantly increased in the OVX+EA group, as compared with the OVX group (p<0.01 and p<0.05, respectively). CONCLUSION: This study demonstrates that EA can prevent OVX-induced bone loss and deterioration of bone architecture and strength by stimulating the Wnt/ß-catenin signalling pathway. These findings suggest that EA may bet a promising adjunct method for inhibiting OVX-induced osteoporosis in clinical settings.

Effects of pulsed electromagnetic fields on bone mass and Wnt/ß-catenin signaling pathway in ovariectomized rats.

BACKGROUND AND AIMS: The therapeutic effects of pulsed electromagnetic fields (PEMFs) on osteoporosis have been documented. However, the precise mechanisms by which PEMFs elicit these favorable biological responses are still not fully understood. This study aimed to systematically investigate the effects of PEMFs on bone mass and Wnt/ß-catenin signaling pathway in ovariectomized rats. METHODS: Thirty 3-month-old female Sprague Dawley rats were randomly assigned to one of three groups: sham-operated control (sham), ovariectomy (OVX), and ovariectomy with PEMFs treatment (PEMFs). One week following ovariectomy surgery, rats in the PEMFs group were exposed to PEMFs for 40 min/day, 5 days/week, for 12 weeks. RESULTS: After 12-week interventions, serum 17ß-estradiol and bone-specific alkaline phosphatase levels increased in the PEMFs group. Bone mineral density of the femur and the fifth lumbar vertebral body also increased in the PEMFs group. Histomorphometrical studies showed that PEMFs improved trabecular area, trabecular width, and trabecular number by 77.50%, 17.38% and 51.06%, respectively, and reduced trabecular separation by 44.28% compared with the OVX group. Biomechanical studies showed that PEMFs increased maximum load and energy to failure in the fifth lumbar vertebral body. Quantitative real-time RT-PCR analysis showed that PEMFs increased the mRNA expressions of Wnt3a, low-density lipoprotein receptor-related protein 5(LRP5), ß-catenin, c-myc and runt-related gene 2 (Runx2), and reduced dickkopf1 (DKK1) in ovariectomized rats. However, mRNA expression of Axin2 was not affected by PEMFs. CONCLUSIONS: PEMFs can prevent ovariectomy-induced bone loss and deterioration of bone microarchitecture and strength, at least partly, through activation of Wnt/ß-catenin signaling pathway.