Phytochemical Investigation of Lepionurus sylvestris Blume and Their Anti-Diabetes Effects via Anti-Alpha Glucosidase and Insulin Secretagogue Activities Plus Molecular Docking.

This study presents a phytochemical investigation of Lepionurus sylvestris leaf extracts and their anti-diabetic activities. Traditionally, L. sylvestris leaves were used as vegetables and food in local recipes, but the root extracts of the plant can also be used in body tonic and erectile dysfunction treatments. Following a preliminary anti-diabetic activity screening test, the 80% ethanolic leaf extract exhibited potent anti-alpha glucosidase activity. So, the leaves' active components were selected for further investigation. Firstly, the plant was extracted via maceration using lower to higher polarity solvents such as hexane, ethyl acetate, ethanol, and water, respectively, to obtain the four crude extracts. Then, the phytochemicals contained in this plant were investigated via classical column chromatography and spectroscopy techniques. Anti-diabetic activity was evaluated via anti-alpha glucosidase and insulin secretagogue assays. The results showed that five compounds were isolated from the fractionated ethanolic leaf extract: interruptin A; interruptin C; ergosterol; diglycerol; and 15-16-epoxy-neo-cleoda-3,7(20),13(16),14-tetraene-12,17:18,19-diolide, a new diterpene derivative which is herein referred to as lepionurodiolide. Interruptin A and the new diterpene derivative exhibited the greatest effect on anti-alpha glucosidase activity, showing IC50 values of 293.05 and 203.71 µg/mL, respectively. Then, molecular docking was used to study the sites of action of these compounds. The results showed that interruptin A and the new compound interacted through H-bonds with the GLN279 residue, with a binding energy of -9.8 kcal/mol, whereas interruptin A and C interacted with HIS280 and ARG315 a with binding energy of -10.2 kcal/mol. Moreover, the extracts were investigated for their toxicity toward human cancer cells, and a zebrafish embryonic toxicity model was used to determine herbal drug safety. The results indicated that ethyl acetate and hexane extracts showed cytotoxicity to both Hela cells and human breast adenocarcinomas (MCF-7), which was related to the results derived from using the zebrafish embryonic toxicity model. The hexane and ethyl acetate presented LC50 values of 33.25 and 36.55 µg/mL, respectively, whereas the ethanol and water extracts did not show embryonic toxicity. This study is the first of its kind to report on the chemical constituents and anti-diabetic activity of L. sylvestris, the leaf extract of which has been traditionally used in southern Thailand as a herbal medicine and food ingredient.

Amelioration of Sensorineural Hearing Loss through Regulation of Trpv1, Cacna1h, and Ngf Gene Expression by a Combination of Cuscutae Semen and Rehmanniae Radix Preparata.

Sensorineural hearing loss (SNHL) is a common condition that results from the loss of function of hair cells, which are responsible for converting sound into electrical signals within the cochlea and auditory nerve. Despite the prevalence of SNHL, a universally effective treatment has yet to be approved. To address this absence, the present study aimed to investigate the potential therapeutic effects of TS, a combination of Cuscutae Semen and Rehmanniae Radix Preparata. To this end, both in vitro and in vivo experiments were performed to evaluate the efficacy of TS with respect to SNHL. The results showed that TS was able to protect against ototoxic neomycin-induced damage in both HEI-OC1 cells and otic hair cells in zebrafish. Furthermore, in images obtained using scanning electron microscopy (SEM), an increase in the number of kinocilia, which was prompted by the TS treatment, was observed in the zebrafish larvae. In a noise-induced hearing loss (NIHL) mouse model, TS improved hearing thresholds as determined by the auditory brainstem response (ABR) test. Additionally, TS was found to regulate several genes related to hearing loss, including Trpv1, Cacna1h, and Ngf, as determined by quantitative real-time polymerase chain reaction (RT-PCR) analysis. In conclusion, the findings of this study suggest that TS holds promise as a potential treatment for sensorineural hearing loss. Further research is necessary to confirm these results and evaluate the safety and efficacy of TS in a clinical setting.

In Vitro, In Vivo, and In Silico Analyses of Molecular Anti-Pigmentation Mechanisms of Selected Thai Rejuvenating Remedy and Bioactive Metabolites.

Thai rejuvenating remedies are mixed herbal formulas promoting longevity. Due to the complexity, the biological activities of these remedies are minimal. Therefore, in this study, the authors evaluated the anti-pigmentation effect at the molecular level of the selected Thai rejuvenating remedy to fulfill the knowledge gap. First, the authors found that the selected remedy showed promising activity against the tyrosinase enzyme with an IC50 value of 9.41 µg/mL. In the comparison, kojic acid (positive control) exhibited an IC50 value of 3.92 µg/mL against the same enzyme. Later, the authors identified glabridin as a bioactive molecule against tyrosinase with an IC50 value of 0.08 µg/mL. However, ethyl p-methoxycinnamate was the most abundant metabolite found in the remedy. The authors also found that the selected remedy and glabridin reduced the melanin content in the cell-based assay (B16F1) but not in the zebrafish larvae experiment. Finally, the authors conducted a computational investigation through molecular docking proposing a theoretical molecular interplay between glabridin, ethyl p-methoxycinnamate, and target proteins (tyrosinase and melanocortin-1 receptor, MC1R). Hence, in this study, the authors reported the molecular anti-pigmentation mechanism of the selected Thai rejuvenating remedy for the first time by combining the results from in silico, in vitro, and in vivo experiments.

The Effects of Persimmon (Diospyros kaki L.f.) Oligosaccharides on Features of the Metabolic Syndrome in Zebrafish.

Metabolic syndrome has become a global health care problem since it is rapidly increasing worldwide. The search for alternative natural supplements may have potential benefits for obesity and diabetes patients. Diospyros kaki fruit extract and its oligosaccharides, including gentiobiose, melibiose, and raffinose, were examined for their anti-insulin resistance and obesity-preventing effect in zebrafish larvae. The results show that D. kaki oligosaccharides improved insulin resistance and high-fat-diet-induced obesity in zebrafish larvae, evidenced by enhanced ß-cell recovery, decreased abdominal size, and reduced the lipid accumulation. The mechanism of the oligosaccharides, molecular docking, and enzyme activities of PTP1B were investigated. Three of the oligosaccharides had a binding interaction with the catalytic active sites of PTP1B, but did not show inhibitory effects in an enzyme assay. The catalytic residues of PTP1B were typically conserved and the cellular penetration of the cell membrane was necessary for the inhibitors. The results of the mechanism of action study indicate that D. kaki fruit extract and its oligosaccharides affected gene expression changes in inflammation- (TNF-α, IL-6, and IL-1ß), lipogenesis- (SREBF1 and FASN), and lipid-lowering (CPT1A)-related genes. Therefore, D. kaki fruit extract and its oligosaccharides may have a great potential for applications in metabolic syndrome drug development and dietary supplements.

Characteristics of the New Insulin-Resistant Zebrafish Model.

Insulin resistance, which occurs when insulin levels are sufficiently high over a prolonged period, causing the cells to fail to respond normally to the hormone. As a system for insulin resistance and diabetes drug development, insulin-resistant rodent models have been clearly established, but there is a limitation to high-throughput drug screening. Recently, zebrafish have been identified as an excellent system for drug discovery and identification of therapeutic targets, but studies on insulin resistance models have not been extensively performed. Therefore, we aimed to make a rapid insulin-resistant zebrafish model that complements the existing rodent models. To establish this model, zebrafish were treated with 10 µM insulin for 48 h. This model showed characteristics of insulin-resistant disease such as damaged pancreatic islets. Then we confirmed the recovery of the pancreatic islets after pioglitazone treatment. In addition, it was found that insulin-resistant drugs have as significant an effect in zebrafish as in humans, and these results proved the value of the zebrafish insulin resistance model for drug selection. In addition, RNA sequencing was performed to elucidate the mechanism involved. KEGG pathway enrichment analysis of differentially expressed genes showed that insulin resistance altered gene expression due to the MAPK signaling and calcium signaling pathways. This model demonstrates the utility of the zebrafish model for drug testing and drug discovery in insulin resistance and diabetes.

Anti-insulin resistance effect of constituents from Senna siamea on zebrafish model, its molecular docking, and structure-activity relationships.

Senna siamea has been used as an antidiabetic drug since antiquity. With regard to traditional Thai medicine, the use of S. siamea was described for diabetes therapy. To understand the molecular mechanism regarding insulin resistance. Pure compounds were isolated from wood extract. We studied their biological activities on insulin-resistance using an in vivo zebrafish model. We also performed an in silico study; molecular docking, and in vitro study by taking advantage of the enzyme inhibitory activities of α-glucosidase, PTP1B, and DPP-IV. Based on the preliminary investigation that ethyl acetate and ethanol extracts have potent effects against insulin resistance on zebrafish larvae, five compounds were isolated from two fractions following: resveratrol, piceatannol, dihydropiceatannol, chrysophanol, and emodin. All of the isolated compounds had anti-insulin resistance effects on zebrafish larvae. Resveratrol, piceatannol, and dihydropiceatannol also demonstrated inhibitory effects against α-glucosidase. Chrysophanol and emodin inhibited PTP1B activity, while resveratrol showed a DPP-IV inhibition effect via the molecular docking. The results of enzyme assay were similar. In conclusions, S. siamea components demonstrated effects against insulin resistance. The chemical structure displayed identical biological activity to that of the compounds. Therefore, S. siamea wood extract and their components are potential therapeutic options in the treatment of diabetes.

Anti-aging effects of Korean Red Ginseng (KRG) in differentiated embryo chondrocyte (DEC) knockout mice.

BACKGROUND: The circadian rhythm is the internal clock that controls sleep-wake cycles, metabolism, cognition, and several processes in the body, and its disruption has been associated with aging. The differentiated embryo chondrocyte (Dec) gene is related to circadian rhythm. To our knowledge, there are no reports of the relationship between dec gene expression and KRG effect. Therefore, we treated Dec gene knockout (KO) aging mice with KRG to study anti-aging related effects and possible mechanisms. METHODS: We evaluated KRG and expression of Dec genes in an ototoxicity model. Dec genes expression in livers of aging mice was further analyzed. Then, we assessed the effects of DEC KO on hearing function in mice by ABR. Finally, we performed DNA microarray to identify KRG-related gene expression changes in mouse liver and assessed the results using KEGG analysis. RESULTS: KRG decreased the expression of Dec genes in ototoxicity model, which may contribute to its anti-aging efficacy. Moreover, KRG suppressed Dec genes expression in liver of wild type indicating inhibition of senescence. ABR test indicated that KRG improved auditory function in aging mouse, demonstrating KRG efficacy on aging related diseases. CONCLUSION: Finally, in KEGG analysis of 238 genes that were activated and 158 that were inhibited by KRG in DEC KO mice, activated genes were involved in proliferation signaling, mineral absorption, and PPAR signaling whereas the inhibited genes were involved in arachidonic acid metabolism and peroxisomes. Our data indicate that inhibition of senescence-related Dec genes may explain the anti-aging efficacy of KRG.

Avocado Oil Extract Modulates Auditory Hair Cell Function through the Regulation of Amino Acid Biosynthesis Genes.

Sensorineural hearing loss (SNHL) is one of the most common causes of disability, affecting over 466 million people worldwide. However, prevention or therapy of SNHL has not been widely studied. Avocado oil has shown many health benefits but it has not yet been studied in regards to SNHL. Therefore, we aimed to investigate the efficacy of avocado oil on SNHL in vitro and in vivo and elucidate its mode of action. For the present study, we used enhanced functional avocado oil extract (DKB122). DKB122 led to recovery of otic hair cells in zebrafish after neomycin-induced otic cell damage. Also, DKB122 improved auditory sensory transmission function in a mouse model of noise induced-hearing loss and protected sensory hair cells in the cochlea. In addition, RNA sequencing was performed to elucidate the mechanism involved. KEGG pathway enrichment analysis of differentially expressed genes showed that DKB122 protected House Ear Institute-Organ of Corti 1 (HEI-OC1) cells against neomycin-related alterations in gene expression due to oxidative stress, cytokine production and protein synthesis.

Anti-Streptococcus mutans efficacy of Thai herbal formula used as a remedy for dental caries.

CONTEXT: Traditional knowledge of herbal remedies plays an important role in the search for more effective alternative treatment of a variety of disorders. The ethnobotanical surveys in southern Thailand have revealed that 35 Thai herbal formulas have been used by Thai traditional healers against dental caries. However, the scientific evaluation to confirm their rational uses is scarce. OBJECTIVE: To test in vitro anti-Streptococcus mutans activity of Thai herbal formulas used against dental caries (THF-DC). MATERIALS AND METHODS: Ethanol extracts of Thai herbal formulas were evaluated for antibacterial activity against S. mutans. Agar disc diffusion was employed as a preliminary screening assay, followed by broth microdilution assay to assess minimum inhibitory concentration (MIC). Furthermore, medicinal plants contained in the most active THF-DC were investigated for their phytochemicals. RESULTS: Eleven THF-DC extracts exhibited clear inhibition zones of 7.0-22.5 mm against S. mutans. Subsequent determination of their MIC revealed that the formula containing Albizia myriophylla Benth. (Leguminosae), Alpinia galanga (L.) Willd. (Zingiberaceae), Avicennia marina (Forssk.) Vierh. (Acanthaceae), and Ocimum sanctum L. (Lamiaceae) was the most active, with MIC at 250 µg/mL. Among these medicinal plants, A. myriophylla gave the strongest activity with MIC at 3.9 µg/mL, followed by A. marina with MIC at 62.5 µg/mL. Various classes of bioactive phytochemicals including tannins, flavonoids, alkaloids, and terpenoids were found in these extracts. CONCLUSION: Anti-S. mutans activity of THF-DC extracts was established. Further investigations may be required for the isolation and chemical characterization of the active ingredients in A. myriophylla.