Licochalcone A and B enhance muscle proliferation and differentiation by regulating Myostatin.

BACKGROUND: Myostatin (MSTN) inhibition has demonstrated promise for the treatment of diseases associated with muscle loss. In a previous study, we discovered that Glycyrrhiza uralensis (G. uralensis) crude water extract (CWE) inhibits MSTN expression while promoting myogenesis. Furthermore, three specific compounds of G. uralensis, namely liquiritigenin, tetrahydroxymethoxychalcone, and Licochalcone B (Lic B), were found to promote myoblast proliferation and differentiation, as well as accelerate the regeneration of injured muscle tissue. PURPOSE: The purpose of this study was to build on our previous findings on G. uralensis and demonstrate the potential of its two components, Licochalcone A (Lic A) and Lic B, in muscle mass regulation (by inhibiting MSTN), aging and muscle formation. METHODS: G. uralensis, Lic A, and Lic B were evaluated thoroughly using in silico, in vitro and in vivo approaches. In silico analyses included molecular docking, and dynamics simulations of these compounds with MSTN. Protein-protein docking was carried out for MSTN, as well as for the docked complex of MSTN-Lic with its receptor, activin type IIB receptor (ACVRIIB). Subsequent in vitro studies used C2C12 cell lines and primary mouse muscle stem cells to acess the cell proliferation and differentiation of normal and aged cells, levels of MSTN, Atrogin 1, and MuRF1, and plasma MSTN concentrations, employing techniques such as western blotting, immunohistochemistry, immunocytochemistry, cell proliferation and differentiation assays, and real-time RT-PCR. Furthermore, in vivo experiments using mouse models focused on measuring muscle fiber diameters. RESULTS: CWE of G. uralensis and two of its components, namely Lic A and B, promote myoblast proliferation and differentiation by inhibiting MSTN and reducing Atrogin1 and MuRF1 expressions and MSTN protein concentration in serum. In silico interaction analysis revealed that Lic A (binding energy -6.9 Kcal/mol) and B (binding energy -5.9 Kcal/mol) bind to MSTN and reduce binding between it and ACVRIIB, thereby inhibiting downstream signaling. The experimental analysis, which involved both in vitro and in vivo studies, demonstrated that the levels of MSTN, Atrogin 1, and MuRF1 were decreased when G. uralensis CWE, Lic A, or Lic B were administered into mice or treated in the mouse primary muscle satellite cells (MSCs) and C2C12 myoblasts. The diameters of muscle fibers increased in orally treated mice, and the differentiation and proliferation of C2C12 cells were enhanced. G. uralensis CWE, Lic A, and Lic B also promoted cell proliferation in aged cells, suggesting that they may have anti-muslce aging properties. They also reduced the expression and phosphorylation of SMAD2 and SMAD3 (MSTN downstream effectors), adding to the evidence that MSTN is inhibited. CONCLUSION: These findings suggest that CWE and its active constituents Lic A and Lic B have anti-mauscle aging potential. They also have the potential to be used as natural inhibitors of MSTN and as therapeutic options for disorders associated with muscle atrophy.

Identification and Evaluation of Traditional Chinese Medicine Natural Compounds as Potential Myostatin Inhibitors: An In Silico Approach.

Myostatin (MSTN), a negative regulator of muscle mass, is reported to be increased in conditions linked with muscle atrophy, sarcopenia, and other muscle-related diseases. Most pharmacologic approaches that treat muscle disorders are ineffective, emphasizing the emergence of MSTN inhibition. In this study, we used computational screening to uncover natural small bioactive inhibitors from the Traditional Chinese Medicine database (~38,000 compounds) for the MSTN protein. Potential ligands were screened, based on binding affinity (150), physicochemical (53) and ADMET properties (17). We found two hits (ZINC85592908 and ZINC85511481) with high binding affinity and specificity, and their binding patterns with MSTN protein. In addition, molecular dynamic simulations were run on each complex to better understand the interaction mechanism of MSTN with the control (curcumin) and the hit compounds (ZINC85592908 and ZINC85511481). We determined that the hits bind to the active pocket site (Helix region) and trigger conformational changes in the MSTN protein. Since the stability of the ZINC85592908 compound was greater than the MSTN control, we believe that ZINC85592908 has therapeutic potential against the MSTN protein and may hinder downstream singling by inhibiting the MSTN protein and increasing myogenesis in the skeletal muscle tissues.

Effects of polyphenol-rich traditional herbal teas on obesity and oxidative stress in rats fed a high-fat-sugar diet.

Hibiscus rosa-sinensis and Zingiber officinalis teas are traditionally used for the therapies of various diseases, including obesity. The present research work was planned to appraise the potential of polyphenol-rich extracts of selected herbal plants in obesity and related biochemical parameters of high-fat-sugar diet-induced obese rats. Three herbal teas were prepared from Hibiscus rosa-sinensis flowers and Zingiber officinalis rhizomes and their mixture (3:1, respectively). Total phenolic contents (TPC) of Hibiscus rosa-sinensis and Zingiber officinalis extracts were found to be 5.82 and 1.45 mg/g of dry plant material, measured as GAE, while total flavonoid contents (TFC) were 9.17 and 1.95 mg/g of dry plant material, measured as CE, respectively. Two doses (250 and 500 mg/kg BW) of each tea were administered and body weight, BMI, kidney, liver, and atherogenic indices, TC, TG, HDL, LDL, VLDL, BT, AST, ALT, AP, SC, MDA, SOD, GSH, and TAC of rats groups were measured. Data showed that higher doses of Hibiscus rosa-sinensis significantly reduced the rat's BMI (0.50 g/cm2) in comparison with the high-fat-sugar diet group (0.79 g/cm2). All treatment groups, especially H-500 group, showed a significant decrease in the elevated kidney and liver weights and atherogenic index in comparison with HFSDC groups. Higher doses of Hibiscus rosa-sinensis significantly decreased the levels of AST, ALT, AP, and SC in comparison with the HFSDC group. A significant decrease in the levels of serum TC, TG, LDL, and VLDL was observed in all the treatment groups in comparison with the HFSDC group. Furthermore, all the teas, especially higher doses of Hibiscus rosa-sinensis, prevented the alterations in MDA, SOD, and GSH levels of experimental groups, thus showing the potential against oxidative stress. It can be concluded from these results that Hibiscus rosa-sinensis teas exhibited strong protective effects against obesity and oxidative stress, especially at higher doses.

Fagonia arabica L.: A Review of its Phytochemistry, Pharmacology and Traditional Uses.

BACKGROUND: Fagonia arabica, belonging to family Zygophyllaceae, is a medicinal plant widely distributed in the desert areas of the world, including Saudi Arabia, Pakistan, India and South Africa. The present review aims to explore the published information on the traditional uses, ethnobotanical knowledge, phytochemistry and various biological activities like antioxidant, antimicrobial, thrombolytic activities and anticoagulant effects of Fagonia arabica with critical analysis on the gaps and future perspectives. METHOD: A literature survey was performed by searching the digital libraries and the scientific databases including Scopus, Google Scholar, SciFinder, ACS, Web of Science and published books. RESULTS: Fagonia arabica plant has been reported to have a wide range of traditional uses in sore mouth, smallpox, hematological, neurological, endocrinological, inflammatory, cooling agent in stomatitis, vertigo and endothermic reaction in the body. Several bioactive constituents, including glycosides, flavonoids, terpenoids, saponins, alkaloids and trace elements, were recorded from Fagonia arabica plant. The isolation and identification of two flavonoid glycosides (kaempferol-7- O-rhamnoside and acacetin-7-O-rhamnoside) were also reported. Fagonia arabica has been studied for its wide range of biological activities, which include antioxidant, antimicrobial, cardioprotective and anticoagulant. CONCLUSION: It is apparent from the literature that Fagonia arabica plant possesses a wide range of medicinal and pharmacological uses and has been studied for its various pharmacological activities and medicinal applications. Critical analysis reveals that the plant has huge potential for pharmaceutical and pharmacological applications.

Catechins, theaflavins and ginger freeze-dried extract based functional drink significantly mitigate the hepatic, diabetic and lipid abnormalities in rat model.

The Current study was planned to explore the therapeutic potential of green tea, black tea and ginger based nutraceuticals (catechins, theaflavins and ginger freeze dried extract) against obesity, diabetes and renal malfunctioning. Bioevaluation study was carried out by involving 250 male Sprague Dawley rats. Accordingly, three types of studies were conducted on the basis of different diets i.e. study I (Hyperglycemic rats), study II (obese rats), study III (liver malfunctional rats) each study comprised of five groups of rats ten in each (Sample size according to power analysis) were provided the five types of drinks i.e. control, theaflavin enriched, catechins enriched, ginger extract supplemented and combination of catechins, theaflavins and ginger extract were given to the representative groups. Results showed that the body weight of rats effected significantly with functional drinks in all studies. However, catechin enriched drink (T1) resulted maximum reduction in weight during the entire study. Similarly, T2 exerted maximum decline in cholesterol level during study I, II and III by 11.03 & 10.63, 7.62 & 8.05 and 5.99 & 6.01% whereas LDL by 14.25 & 15.10, 10.45 & 12.10 and 7.25 & 8.01%, respectively (trial 1 & 2). The attenuation in serum glucose and enhancement in insulin level of rats are the indicators for the positive impact of black tea functional drinks. In this context, Catechins+theaflavins+GFD enriched drink (T4) Showed better performance than rest and caused 8.82 & 9.77, 11.03 & 12.23 and 5.83 & 5.96% reduction in glucose. Moreover, the T4 significantly improved the liver and antioxidant enzymes. Accordingly, T4 was proved effective for glutathione enhancement whilst T2 alleviated TBARS efficiently during the investigation. The normal ranges of renal function tests and hematological aspects proved the safety of resultant drinks. From the current exploration, it is concluded that drinks supplemented with theaflavin and catechins & GFD are effectual to mitigate lifestyle related malfunctioning.

Comparative genomic analysis of human GLI2 locus using slowly evolving fish revealed the ancestral gnathostome set of early developmental enhancers.

BACKGROUND: The zinc finger-containing transcription factor Gli2, is a key mediator of Hedgehog (Hh) signaling and participates in embryonic patterning of various organs including the central nervous system (CNS) and limbs. Abnormal expression of Gli2 can impede the transcription of Hh target genes through disruption of proper balance between Gli2 and Gli3 functions. Therefore, delineation of enhancers that are required for complementary roles of Glis would allow the interrogation of those pathogenic variants that cause gene dysregulation, and a corresponding abnormal phenotype. Previously, we reported tissue-specific enhancers for Gli family including Gli2 through direct tetrapod-teleost comparisons. RESULTS: Here, we employed the sequence alignments of slowly evolving spotted gar and elephant shark and have identified six novel conserved noncoding elements in human GLI2 containing locus. Zebrafish-based transgenic assays revealed that combined action of these autonomous CNEs reflects many aspects of Gli2 specific endogenous transcriptional activity, including CNS and pectoral fins. CONCLUSION: Taken together with our previous findings, this study suggests that Hh-signaling controlled deployment of Gli2 activity in embryonic patterning arose in the common ancestor of gnathostomes. These GLI2 specific cis-regulatory modules will help to identify DNA variants that probably reside outside of coding intervals and are associated with congenital anomalies.

Identification and Evaluation of Inhibitors of Lipase from Malassezia restricta using Virtual High-Throughput Screening and Molecular Dynamics Studies.

Recent studies revealed the role of lipase in the pathogenicity of Malassezia restricta in dandruff and seborrheic dermatitis (D/SD). The lipase from M. restricta (Mrlip1) is considered a potential target for dandruff therapy. In this work, we performed structure-based virtual screening in Zinc database to find the natural bioactive inhibitors of Mrlip1. We identified three compounds bearing superior affinity and specificity from the Traditional Chinese Medicine database (~60,000 compounds), and their binding patterns with Mrlip1 were analyzed in detail. Additionally, we performed three sets of 100 ns MD simulations of each complex in order to understand the interaction mechanism of Mrlip1 with known inhibitor RHC80267 and the newly identified compounds such as ZINC85530919, ZINC95914464 and ZINC85530320, respectively. These compounds bind to the active site cavity and cause conformational changes in Mrlip1. The Molecular Mechanics Poisson-Boltzmann Surface Area (MMPBSA) studies suggested that the average binding energy was stronger in the case of Mrlip1-ZINC85530919 and Mrlip1-ZINC95914464. The selected natural inhibitors might act as promising lead drugs against Mrlip1. Further, the present study will contribute to various steps involved in developing and creating potent drugs for several skin diseases including dandruff.

Highly Efficient Deacidification of High-Acid Rice Bran Oil Using Methanol as a Novel Acyl Acceptor.

A highly efficient process for reducing the fatty acid (FA) content of high-acid rice bran oil (RBO) was developed by immobilized partial glycerides-selective lipase SMG1-F278N-catalyzed esterification/transesterification using methanol as a novel acyl acceptor. Molecular docking simulation indicated that methanol was much closer to the catalytic serine (Ser-171) compared with ethanol and glycerol, which might be one of the reasons for its high efficiency in the deacidification of high-acid RBO. Additionally, the reaction parameters were optimized to minimize the FA content of high-acid RBO. Under the optimal conditions (substrate molar ratio of methanol to FAs of 1.8:1, enzyme loading of 40 U/g, and at 30 °C), FA content decreased from 25.14 to 0.03% after 6 h of reaction. Immobilized SMG1-F278N exhibited excellent methanol tolerance and retained almost 100% of its initial activity after being used for ten batches. After purification by molecular distillation, the final product contained 97.86% triacylglycerol, 2.10% diacylglycerol, and 0.04% FA. The acid value of the final product was 0.09 mg KOH/g, which reached the grade one standard of edible oil. Overall, methanol was a superior acyl acceptor for the deacidification of high-acid RBO and the high reusability of immobilized SMG1-F278N indicates an economically attractive process.

Variation in Phenolic Profile, ß-Carotene and Flavonoid Contents, Biological Activities of Two Tagetes Species from Pakistani Flora.

The objective of present study was to evaluate the variation in phenolic profile, ß-carotene, flavonoid contents, antioxidant and antimicrobial properties of Tagetes erecta and Tagetes patula (T. erecta and T. patula) through different in vitro assays. Antioxidant activity was determined through 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging and inhibition of linoleic acid peroxidation assays and antibacterial and antifungal activities studied using the disc diffusion and resazurin microtiter-plate assays against bacterial and fungal strains. Moreover, total phenolics (TP), total carotenoids (TC) and total flavonoids (TF) were also determined. Highest (TP 35.8 mg GAE/g) and TF (16.9 mg CE/g) contents were found in MeOH extract of T. patula. T. erecta extract showed higher TC contents (6.45 mg/g) than T. patula extract (6.32 mg/g). T. erecta exhibited the highest DPPH radical-scavenging activity (IC50 ) (5.73 µg/mL) and inhibition of linoleic acid peroxidation (80.1%). RP-HPLC revealed the presence of caffeic acid, sinapic acid and ferulic acid in Tagetes extracts, m-coumaric acid in T. erecta whereas chlorogenic acid in T. patula extract only. Both extracts possessed promising antimicrobial activity compared to the ciprofloxacin and flumequine (+ve controls) against Bacillus subtilis and Alternaria alternate. Both extract were rich source of polyphenols exhibiting excellent biological activities.