The insulin signaling pathway, involving protein kinase B (PKB) and mitogen-activated protein kinase (MAPK), mediates the biological response to insulin and several growth factors and cytokines. To investigate the correlation between glucose transporter (Glut) biosynthesis and the insulin signaling pathway activated by novel compounds of Liriope platyphylla (LP9M80-H), alterations in Glut and key protein expression in the insulin signaling pathway were analyzed in the liver and brain of ICR mice treated with LP9M80-H. An in vitro assay showed that the highest level of insulin concentration was observed in the LP9M80-H-treated group, followed by the LP-H, LP-M, LP-E, and LP9M80-C-treated groups. Therefore, LP9M80-H was selected for use in studying the detailed mechanism of the insulin signaling pathway in animal systems. In an in vivo experiment, LP9M80-H induced a significant increase in glucose levels and a decrease of insulin concentration in the blood of mice, while their body weight remained constant over 5 days. The expression level of Glut-3 was down-regulated in the liver, or maintained at the same level in the brain of LP9MH80-H-treated mice. These changes corresponded to the phosphorylation of the p38 protein rather than to ERK and JNK in the MAPK signaling pathway. In addition, the expression level of Glut-1 increased significantly after LP9MH80-H treatment of both insulin target tissues in mice. Western blot analysis showed that Akt in the PI3-K pathway mainly participated in Glut-1 biosynthesis. Thus, these results suggest the possibility that the LP9M80-H-induced regulation of Glut-1 and Glut-3 biosynthesis may be mediated by the Akt and p38 MAPK signaling of the insulin signaling pathway in the liver and brain of mice.

Although oxidative stress plays a critical role in age-related macular degeneration (AMD) progression, natural product-derived single compounds against AMD remain largely unexplored. We investigated the protective effects and underlying mechanism of α-iso-cubebene against AMD-like retinal injury. Alterations in key phenotypes for AMD were analyzed in AMD-mimicking models using ARPE-19 cells co-treated with blue light (BL) and N-retinylidene-N-retinylethanolamine (A2E), as well as BL-exposed BALB/c mice. In BL+A2E-treated ARPE-19 cells, α-iso-cubebene reduced intracellular reactive oxygen species (ROS) and nitric oxide (NO) production and restored superoxide dismutase (SOD) activity and nuclear factor erythroid 2-related factor 2 (Nrf2), suggesting enhancement of the antioxidant defense system. Furthermore, α-iso-cubebene improved cell viability, reduced apoptotic cell populations, and regulated apoptosis-related signaling pathways under oxidative stress conditions. It also attenuated cyclooxygenase-2 (COX-2)-mediated inducible nitric oxide synthase (iNOS) signaling and was associated with reduced inflammasome-related signaling. Importantly, these protective effects were consistently observed regarding the protection of histopathological structure and normalization of inflammatory cytokines in the retina of BL-exposed BALB/c mice. Collectively, our results demonstrate that α-iso-cubebene, as a potential therapeutic candidate, alleviates AMD-like retinal injury and was associated with enhanced antioxidant responses and reduced inflammatory and apoptotic signaling markers.

Polydopamine (PDA) surface coatings are widely used in biomedical engineering to enhance cell-substrate interactions; however, their effects on cancer-cell behavior remain unclear. In this study, we investigated how PDA-coated two-dimensional (2D) culture surfaces influence oncogenic traits of human prostate cancer (PC) cells in vitro. Using LNCaP, DU145, and PC3 cell lines, we found that PDA-coated substrates markedly increased the adhesion, migration, invasion, proliferation, and colony formation in a dose- and time-dependent manner. PDA exposure also induced epithelial-mesenchymal transition (EMT), upregulated cancer stem cell markers (CD44, CD117, CD133, Sox2, Oct4, and Nanog), and elevated expression of metastasis- and chemoresistance-associated molecules (MMP-2, MMP-9, MDR1, and MRP1). Mechanistically, PDA coatings enhanced integrin α2β1-associated cell adhesion, accompanied by increased focal adhesion kinase (FAK) phosphorylation and downstream activation of JNK signaling. Pharmacological inhibition of integrin α2β1 (BTT-3033), FAK (PF573228) and JNK (SP600125) effectively abrogated PDA-induced malignant phenotypes and restored chemosensitivity to cabazitaxel, cisplatin, docetaxel, curcumin, and enzalutamide. Collectively, these findings identify PDA-coated surfaces as a simple, efficient, and reductionist in vitro platform for studying adhesion-mediated signaling and phenotypic plasticity in PC cells, while acknowledging that further validation in three-dimensional (3D) and patient-derived models will be required to establish in vivo relevance.

In the original publication [...].

BACKGROUND:NKX3.1 is a prostate-specific tumor suppressor that is frequently downregulated during the early stages of prostate cancer. Although NKX3.1 knockout (KO) mice develop spontaneous epithelial abnormalities, these lesions rarely progress beyond early neoplastic changes without additional oncogenic stimulus. Therefore, we investigated whether exogenous testosterone (TS) exacerbates early-stage, pre-neoplastic lesions in the prostate of NKX3.1 KO mice. Alterations in prostate weights of male reproductive organs (testis, seminal vesicles, and prostate lobes), histopathological lesion scores, apoptotic proteins, and angiogenic proteins were analyzed in C57BL/6 NKX3.1em1Hlee/Korl KO (NKX3.1 KO) mice injected with TS for six weeks. RESULTS:The weight of testis, seminal vesicles and ventral prostate was commonly changed in TS-treated mice of wild type (WT) and NKX3.1 KO group, while those of the dorsolateral and anterior prostate were only increased in TS-treated NKX3.1 KO group compared to those of WT. Histopathological lesion severity was greater in TS-treated NKX3.1 KO mice, with the highest lesion scores observed in the high-dose TS (HiTS)-treated KO group, and a similar pattern was observed for p53 staining. The expression levels of apoptotic and angiogenic proteins were significantly increased in TS-treated NKX3.1 KO mice compared to the same group of WT mice. CONCLUSIONS:These findings suggest that the exogenous TS exacerbates early-stage, pre-neoplastic lesions in the prostate of NKX3.1 KO mice, consistent with a gene-hormone synergistic interaction. The mechanistic basis of this synergy remains to be defined.

To identify and characterize the novel odor markers associated with aging in mice, alterations in the concentration of odorants in the urine, expression of metabolic enzymes, the histopathological structure of the sweat glands, and the expression of regulatory factors in sweat secretion were analyzed in ICR mice of four different ages (2, 6, 8 and 10-month-old). The concentrations of 15 odorants including ethylenimine and trimethylamine (TMA), and total volatile organic compounds (VOCs) were significantly higher in the urine of 8- and 10-month-old ICR mice. Among them, TMA was selected as an important odor biomarker associated with aging based on the aging-related upregulation and its odor characteristics. Also, the increase in TMA concentrations was reflected in the transcription levels of the gene encoding TMA monooxygenase in the livers and the feet as well as an age-dependent increase in the lumen area and secretory coiled portion of the sweat glands of mice. These results provide novel scientific evidence that the age-dependent changes in the volatiles in the urine are indicative of aging-related odors in ICR mice. Specifically, the study shows that TMA has potential as a novel diagnostic odor biomarker associated with aging in ICR mice to establish experimental animal platform for analyzing the efficacy and action mechanism of deodorants.

The inhalation effects of airborne nanoplastics (NPs) on the gastrointestinal (GI) tract are rarely investigated, with most studies focusing only on the lungs and other organs. This study examined the changes in the key constipation phenotypes, mucin secretion, water and ion balance, and enteric nervous system (ENS) function in ICR mice after inhalation of polystyrene (PS)-NPs with 500 nm size for two weeks to determine if inhaled NPs can cause constipation. Significant constipation phenotypes, including the weight, water contents and morphology of stools, GI motility, intestinal length, histopathological structure of the colon, and concentration of GI hormone, were detected in NPs-inhaled ICR mice compared to a Vehicle-treated group. In addition, NPs-induced defecation delay was accompanied by a decrease in mucin secretion, suppressed transcription of mucin-related genes, and abnormal regulation of the colonic fluid transport system, including water and ions. NPs-inhaled mice showed a decrease in the neuronal cell density and dysfunction of excitatory neurons and inhibitory neurons in the ENS of the colon. Moreover, similar phenotypes for constipation were verified in ICR mice injected intravenously with NPs. Therefore, these results provide the first scientific evidence that inhalation and intravenous injection of PS-NPs can be considered as one of novel causes of constipation.

Background: Obesity is a global health issue closely associated with dysregulated lipid metabolism and chronic inflammation. Effective strategies targeting both lipogenesis and inflammation are essential for managing obesity and its related metabolic disorders. Methods: This study evaluated the effects of Terminalia catappa Linn. leaf extract (TCE) on lipogenic and lipolytic pathways in high-fat diet (HFD)-induced obese mice. UPLC-QTOF-MS analysis was conducted to identify and quantify the major phenolic compounds in TCE. Mice were administered low and high doses of TCE, and various metabolic parameters, including lipid profiles, liver function markers, adipokine levels, and gene/protein expressions related to lipid metabolism and inflammation, were assessed. Results: UPLC-QTOF-MS analysis identified four major phenolic compounds in TCE—gallic acid, orientin, vitexin, and ellagic acid—with respective contents of 112.5, 163.3, 184.7, and 295.7 mg/g extract. TCE administration significantly reduced liver and adipose tissue weights, along with hepatic and adipose lipid accumulation. Both low and high doses of TCE markedly lowered serum lipid levels. Liver function was improved, as indicated by reduced levels of AST, ALT, and ALP, while BUN levels remained unchanged. On the molecular level, TCE downregulated adipogenic and lipogenic genes (PPARγ, PPARα, C/EBPα, aP2) and upregulated metabolic regulators, including leptin, adiponectin, p-HSL/HSL, and p-perilipin/perilipin, without affecting ATGL expression. TCE also suppressed pro-inflammatory cytokines such as IL-6, IL-1β, TNF-α, and TGFβ-1. Conclusions: These findings highlight the therapeutic potential of TCE in managing obesity by inhibiting lipogenesis, enhancing lipolysis, and reducing inflammation.

Oxidative stress is the key cause of the etiopathogenesis of several diseases associated with constipation. This study examined whether the green pine cone can improve the symptoms of constipation based on the antioxidant activities. The changes in the key parameters for the antioxidant activity and laxative effects were examined in the loperamide (Lop)-induced constipation of Sprague-Dawley (SD) rats after being treated with the methanol extracts of green pine cone (MPC, unripe fruits of Pinus densiflora). MPC contained several bioactive compounds, including diterpenoid compounds such as dehydroabietic acid, taxodone, and ferruginol. In addition, it exhibited high scavenging activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radicals. These effects of MPC successfully reflected the improvement in nicotinamide adenine dinucleotide phosphate oxidase (NADP) H oxidase transcription, superoxide dismutase (SOD) levels, and nuclear factor erythroid 2-related factor 2 (Nrf2) phosphorylation levels in the mid colon of Lop+MPC-treated SD rats. Furthermore, significant improvements in the stool parameters, gastrointestinal (GI) transit, intestine length, and histopathological structure of the mid colon were detected in the Lop-induced constipation rats after MPC treatment. The other parameters, including the regulators for the adherens junction (AJ) and tight junction (TJ), and GI hormone secretion for laxative effects, were improved significantly in Lop+MPC-treated SD rats. These effects were also verified in Lop+MPC-treated primary rat intestine smooth muscle cells (pRISMCs) through analyses for antioxidant defense mechanisms. Overall, the finding of this study offers novel scientific evidence that MPC could be considered as a significant laxative for chronic constipation based on its antioxidant activity.

Background/Objectives: Constipation can be induced in animal models through various factors such as loperamide (Lop) or complement component 3 (C3) deficiency. The effectiveness of therapeutic agents in the clinical management of constipation has been primarily evaluated within only one model, but between-model comparisons have not been performed so far. Therefore, we investigated whether the effectiveness of the laxative drugs for the clinical management is related to etiological factors. Methods: The changes in the key parameters for defecation were compared between C3 knockout (KO) mice with C3-deficiency-induced constipation and ICR mice with Lop-induced constipation after the oral administration of Uridine (Urd) and aqueous extract of Liriope platyphylla L. (AELP). Results: Similar effectiveness of Urd and AELP were detected on the stool frequency, intestinal epithelial barrier structure, and mucin secretion in both models. However, other parameters (namely gastrointestinal (GI) transit, water retention, and enteric nervous system (ENS) structure and function) showed higher effectiveness in C3 KO mice than in the Lop-induced model. Only the effectiveness of the two therapeutic agents on the histological structure of the mid-colon was greater in the Lop-induced mice model compared to the C3 KO mice model. Furthermore, these differences in the therapeutic effectiveness of Urd and AELP were partially reflected in alterations in the cyclic adenosine monophosphate (cAMP) downstream signaling pathway. Conclusions: The results suggest that the therapeutic effectiveness of Urd and AELP is sensitive to C3-deficiency-induced constipation and these differences may be linked to the alternative regulation of the cAMP downstream signaling.

The efficacy of Lagerstroemia speciosa (banaba) leaf extract (BLE), policosanol (POL), and their combination (BLE+POL) was evaluated in zebrafish (Danio rerio) against high cholesterol (HC)- and galactose (HG)-induced metabolic stress and organ toxicity. After 12 weeks of dietary intervention, BLE+POL significantly reduced HC+HG-augmented weight gain and improved hepatic and nephromegaly. Compared with BLE or POL alone, the combined intake of BLE+POL more effectively alleviated dyslipidemia and blood glucose levels. Likewise, BLE+POL effectively reduced blood malondialdehyde (MDA), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) levels and boosted plasma sulfhydryl content, ferric ion reduction ability (FRA), and paraoxonase (PON) activity. Histological outcomes suggest that BLE+POL has higher efficacy than either BLE or POL in mitigating HC+HG-induced fatty liver changes, hepatic inflammation, kidney senescence, and reactive oxygen species (ROS) production. Consistently, BLE+POL augmented the spermatozoa counts in the testes, enhanced mature vitellogenic oocytes in ovaries, and protected them from the HC+HG-induced oxidative stress. Compared with either BLE or POL, a combined intake of BLE+POL displayed a superior effect in inhibiting the apoptosis and accumulation of lipid peroxidation species 4-hyrdoxynonenal (4-HNE) in the brain. A combined intake of BLE+POL exhibited a pronounced impact than the BLE and POL alone and can be utilized as an effective formulation to counteract the HC+HG-induced events.

The internalization mechanism of microplastics (MPs) into human cells has attracted considerable attention because these mechanisms are closely related to the physical and chemical properties of MPs. This study examined the response of human colon cells to autophagy, ER stress, and inflammation during the regulation on the internalization of polystyrene (PS)-MPs (0.4-0.6 μm size). To achieve this, changes in their key markers were analyzed in MPs-treated SNU-1826 cells after a cotreatment with uptake inhibitors or stimulators. The internalization of MPs was significantly higher in SNU-1826 cells than in other cells originated from differential tissues, such as the small intestine, kidneys, and nerves. On the other hand, the internalization of MPs into SNU-1826 cells was suppressed by cytochalasin D (CD) but not by pitstop (Pt). During this inhibition, the levels of the key parameters for autophagy (Light Chain 3-I/II (LC3-I/II) and Beclin1), ER stress (eukaryotic translation initiation factor 2 subunit alpha (EIF2α) and inositol-requiring kinase 1 alpha (IRE1α)), and inflammation (inducible nitric oxide synthase (iNOS), cyclooxygenase 2 (COX-2), tumor necrosis factor alpha (TNF-α), and interleukin (IL)-6) were suppressed in MPs+CD-treated SNU-1826 cells. In addition, the internalization of MPs into SNU-1826 cells was stimulated by a ZnSO4 treatment, not by CaCl2. These stimulation effects were reflected in the alteration of the critical parameters for autophagy, ER stress, and inflammation. Furthermore, the positive correlation was detected between MPs internalization and most parameters for cellular responses although their inhibition is stronger than stimulation. These results suggest that the internalization of MPs into SNU-1826 cells may be strongly associated with the changes in autophagy, ER stress, and inflammation during the regulation of CD and ZnSO4.

This study investigated the anti-obesity and anti-inflammatory effects of D-Allulose compared to erythritol in high-fat diet (HFD)-fed mice, focusing on liver mitochondrial function. Both D-Allulose and erythritol significantly reduced body weight gain, white adipose tissue (WAT) weight, and plasma lipid levels. D-Allulose, especially at higher doses (H_AL), demonstrated superior effects, including reductions in visceral and total WAT weight, adipocyte size, and hepatic and WAT fibrosis, compared to erythritol. D-Allulose also significantly enhanced mitochondrial function, as shown by increased expression of genes and proteins related to lipolysis and oxidative phosphorylation, which were not observed in the erythritol group. Gene expression analysis revealed that D-Allulose more effectively restored HFD-altered gene expression, suggesting stronger regulation of obesity-induced metabolic and inflammatory processes. These findings highlight D-Allulose as a functional sweetener with superior anti-obesity and anti-inflammatory effects, primarily through improved mitochondrial function and modulation of immune and metabolic responses.

This study investigated the levels of sugars and mycotoxins in 41 commercially available fermented vinegars. The validity of the analytical methods was confirmed through evaluation of the limits of detection, limits of quantification, linearity, and recovery rates for both sugars and mycotoxins. The sugar content of the fermented vinegar products showed large differences by region. Among the sugars, glucose and fructose were high predominant. In contrast, sucrose, lactose and maltose were present in smaller amounts, while raffinose and rhamnose were not detected. Among the fermented vinegars, apple vinegar showed a total sugar content that was 5.7 times higher than brown rice vinegar and 7.1 times higher than persimmon vinegar. Regarding mycotoxins, none of the 41 fermented vinegar samples showed detectable levels of aflatoxins B1 · B2 · G1 · G2, ochratoxin A, fumonisin B2, deoxynivalenol and zearalenone. However, fumonisin B1 was detected at a trace level of 1.84±0.03 μg/kg in one apple vinegar sample (V15), which is significantly lower than the maximum limit specified in the Korean Food Code. Therefore, this study provides important data on the sugar and mycotoxin contents of fermented vinegar distributed commercially in Korea.

Skin tissue is susceptible to oxidative stress-induced senescence provoked by ultraviolet (UV) exposure in our daily lives, resulting in photoaging. Herein, we explore whether N-benzyl-N-methyldecan-1-amine (BMDA) derived from garlic ameliorates UVB-induced photoaging. To address this issue, HaCaT keratinocytes were exposed to UVB irradiation under BMDA treatment. The presence of BMDA substantially reduced UVB-induced ROS levels in a dose-dependent manner. BMDA administration counteracted UVB-induced senescence in the β-galactosidase assay. Treatment with BMDA also rescued UVB-exposed cells (S phase; from 18.3 to 25.8%) from cell cycle arrest, similar to the level observed in untreated normal cells. These findings might support our observation that elevated levels of γ-H2AX, a DNA damage marker, under UVB exposure were reduced following BMDA administration. Additionally, BMDA treatment indirectly reduced UVB-induced melanin synthesis in melanocytes since BMDA failed to inhibit tyrosinase activity, a crucial enzyme in melanin synthesis. The topical application of BMDA on the skin of SKH-1 hairless mice also diminished wrinkle formation, supported by recovered collagen levels and the thickness of the epidermis and dermis, compared to those of UVB-control mice. Finally, the BMDA treatment diminished the expression of inflammatory cytokine transcripts such as TNF-α, IL-1β, IL-4, and IL-6 in the UVB-exposed skin tissues. This finding is further supported by Immunofluorescence microscopy, which showed a decrease in the expression of TNF-α, and IL-1β during BMDA treatment. Altogether, as BMDA mitigates UVB-induced photoaging by reducing ROS production, protecting against DNA damage, and suppressing inflammatory cytokine production, it has been proposed as an effective anti-photoaging molecule.

Background: This study aimed to explore the therapeutic potential of a dietary regimen of banaba leaf extract (BNB), policosanol (PCO, Raydel®), and their combination (BNB+PCO), to mitigate high cholesterol (HC) and high galactose (HG) diet-induced dyslipidemia, hyperglycemia, oxidative stress, senescence, and organ damage in zebrafish (Danio rerio). Methodology: Zebrafish (n = 28/group) were fed with HC (4% w/w)+HG (30% w/w) or HC+HG supplemented either with BNB (0.1% w/w) or PCO (0.1% w/w) or BNB+PCO (0.1% w/w each). Following 6 weeks of dietary intervention, biochemical and histopathological examinations across the groups were performed. Results: Post 6 weeks of consumption, the BNB+PCO group exhibited a significant 40% decrease in body weight (BW) relative to the BW of the HC+HG group, while the BNB or PCO groups displayed nonsignificant changes in BW. Both BNB and PCO reduced HC+HG-induced dyslipidemia and hyperglycemia; however, co-administration (BNB+PCO) demonstrated a significantly greater therapeutic effect in countering these conditions compared to either BNB or PCO alone. A similar effect of the BNB+PCO combination was observed on the elevation of plasma sulfhydryl content, paraoxonase (PON), and ferric ion reduction activity (FRA), with notably ~1.2-times (p < 0.01) higher levels compared to their corresponding values observed in the BNB or PCO groups. Significantly diminished plasma AST, ALT, hepatic interleukin 6 (IL-6) levels, and fatty liver changes were observed in response to BNB+PCO, compared to either BNB or PCO alone. Also, BNB+PCO displayed a higher curative effect against HC+HG-induced impairment of tissue regeneration than BNB or PCO alone. A notable effect of BNB+PCO was perceived in protecting kidneys, testis, and ovary damage. Consistently, BNB+PCO showed a profound impact on mitigating HC+HG elevated reactive oxygen species (ROS) generation, apoptosis, cellular senescence, and accumulation of brain-binding lipid proteins (BLBPs) and 4-hydroxynoneal (4-HNE) in the brain. Conclusions: The findings highlight the synergistic effects of the BNB and PCO combination to mitigate the adversity posed by the consumption of the HC+HG diet.

The oral administration of microplastics (MP) for 9 weeks induced disruption of hepatic lipid, glucose, and amino acid metabolism in C57BL/6-Lepem1hwl/Korl (Lep KO) mice with obesity. Therefore, we investigated whether MP-caused hepatic metabolism abnormalities can affect the structural variation of the fecal microbiota during obesity. The overall microbiota profile was analyzed in the feces of Lep KO mice treated with MP for 9 weeks. The lipid accumulation and steatosis area were significantly decreased in MP-treated Lep KO mice. Total microbiota with MP-caused difference identified from feces of Lep KO and wild type (WT) mice were classified into 10 phyla and 106 genera. Among them, two microbial phyla were significantly changed in Lep KO mice after treatment of MP, while significant alterations on 12 genera were detected in Lep KO mice treated with MP. Also, the Chao1 index for richness were remarkably decreased in both MP-treated Lep KO and WT mice, but Shannon index for evenness were increased in only MP-treated Lep KO mice. Therefore, the results of the present study suggest that MP-caused hepatic metabolism disruption may be closely linked to the dysbiosis of the fecal microbiota in Lep KO mice.

The development of physiologically relevant three-dimensional (3D) culture systems is essential for modeling tumor complexity and improving the translational impact of cancer research. We established a 3D in vitro model of human hepatocellular carcinoma (HCC) using a marine collagen peptide-based (MCP-B) biomimetic hydrogel scaffold optimized for multicellular spheroid growth. Compared with conventional two-dimensional (2D) cultures, the MCP-B hydrogel more accurately recapitulated native tumor biology while offering simplicity, reproducibility, bioactivity, and cost efficiency. HCC cells cultured in MCP-B hydrogel displayed tumor-associated behaviors, including enhanced proliferation, colony formation, migration, invasion, and chemoresistance, and enriched cancer stem cell (CSC) populations. Molecular analyses revealed upregulated expression of genes associated with multidrug resistance; stemness regulation and markers; epithelial–mesenchymal transition (EMT) transcription factors, markers, and effectors; growth factors and their receptors; and cancer progression. The spheroids also retained liver-specific functions, suppressed apoptotic signaling, and exhibited extracellular matrix remodeling signatures. Collectively, these findings demonstrate that the 3D HCC model using MCP-B hydrogel recapitulates key hallmarks of tumor biology and provides a robust, physiologically relevant platform for mechanistic studies of HCC and CSC biology. This model further holds translational value for preclinical drug screening and the development of novel anti-HCC and anti-CSC therapeutics.

Background: The efficacy of banaba leaf extract was tested against carboxymethyllysine (CML)-induced toxicity in embryos and adult zebrafish. Additionally, the individual and combined effects of banaba (BNB) and policosanol (PCO) were analyzed to alleviate dyslipidemia, hyperglycemia, and associated effects in streptozotocin (STZ)-induced hyperlipidemic diabetic zebrafish. Methodology: The high cholesterol diet (HCD, final 4%, w/w)-fed zebrafish were injected with STZ to develop diabetes and were subsequently fed with either HCD or HCD+BNB (final 0.1% w/w) or HCD+PCO (final 0.1% w/w) or HCD+BNB+PCO (each final 0.1%, w/w) each for 14 days. The zebrafish tail fin was amputated to assess tissue regeneration, while the organs and blood were collected for histological and biochemical analysis. Results: Severely compromised embryo survivability and developmental defects were noticed in the CML-injected group that significantly improved following BNB exposure. Similarly, CML-induced acute paralysis and mortality of adult zebrafish were effectively mitigated by the treatment with BNB. In the hyperlipidemic diabetic zebrafish, both BNB and PCO supplementation displayed the hypoglycemic effect; however, a remarkable reduction (p < 0.05) in blood glucose levels was observed in the BNB+PCO group, around 14% and 16% less than the BNB group and PCO group, respectively. Likewise, higher tail fin regeneration was noticed in response to BNB+PCO supplementation. Both BNB and PCO have a substantial counter-effect against HCD+STZ-induced dyslipidemia. However, the combined supplementation (BNB+PCO) displayed a significantly better effect than that of BNB and PCO alone to alleviate total cholesterol (TC), triglycerides (TGs), and low-density lipoprotein cholesterol (LDL-C). The most impressive impact of BNB+PCO was noticed in the elevation of high-density lipoprotein cholesterol (HDL-C), which was ~1.5 times higher than the HDL-C level in response to BNB and PCO. Also, BNB+PCO effectively reduced the malondialdehyde (MDA) and elevated the plasma sulfhydryl content, paraoxonase (PON), and ferric ion reduction (FRA) activity. Histological analyses revealed a significant effect of BNB+PCO in preventing inflammatory infiltration, fatty liver changes, and interleukin-6 production. Similarly, a notably better effect of BNB+PCO compared to their individual effect was noticed in preventing kidney damage and mitigation of ROS generation, apoptosis, and cellular senescence. Conclusions: The finding establishes the substantial effect of BNB and PCO in countering hyperglycemia, dyslipidemia, and associated disorders, which synergistically improved following the combined supplementation with BNB+PCO.

ETHNOPHARMACOLOGICAL RELEVANCE:Microsorum membranaceum (D. Don) Ching have a great potential for improving sarcopenia as a form of muscle atrophy because its high antioxidant activity and traditional ailments improving effects. AIM OF THE STUDY:This study aimed to investigate the protective effects and underlying mechanisms of a methanol extract of Microsorum membranaceum (MEM) against dexamethasone-induced sarcopenia. MATERIALS AND METHODS:After analyzing the ingredients and antioxidant properties of MEM, its effects on protein degradation and synthesis pathways, and muscle function were evaluated in dexamethasone (Dex)-treated C2C12 cells and C57BL/6 mice. RESULTS:Fifteen components with bioactivity were detected using HPLC and high scavenging activities against free radicals were measured in MEM. In the protein degradation pathway, MEM suppressed the relative levels of ROS, MuRF1, and Atrogin-1 dose-dependently. In addition, MEM dose-dependently suppressed the expression level of LC3B and Beclin-1 in Dex + MEM-treated C2C12 cells. In the protein synthesis pathway, the MEM treatment induced the enhancement of PI3K, Akt and mTOR phosphorylation as well as the decrease of Myogenin and MyoD transcription in C2C12 cells with Dex-induced sarcopenia. These effects of MEM were well reflected by the increased myotube diameter in the Dex + MEM-treated C2C12 cells. The anti-sarcopenia effects of MEM were successfully verified in Dex-induced sarcopenia of the C57BL/6 mice. Furthermore, Dex + MEM-treated C57BL/6 mice showed significant recovery effects on the weight and function of muscle, as well as the section area of the calf muscle. CONCLUSIONS:These results indicate that MEM can help prevent Dex-induced sarcopenia of C57BL/6 mice by targeting the protein degradation and synthesis pathways.