A comparison of the gene expression profiles of non-alcoholic fatty liver disease between animal models of a high-fat diet and methionine-choline-deficient diet

Non-alcoholic fatty liver disease (NAFLD) embraces several forms of liver disorders involving fat disposition in hepatocytes ranging from simple steatosis to the severe stage, namely, non-alcoholic steatohepatitis (NASH). Recently, several experimental in vivo animal models for NAFLD/NASH have been established. However, no reproducible experimental animal model displays the full spectrum of pathophysiological, histological, molecular, and clinical features associated with human NAFLD/NASH progression. Although methionine-choline-deficient (MCD) diet and high-fat diet (HFD) models can mimic histological and metabolic abnormalities of human disease, respectively, the molecular signaling pathways are extremely important for understanding the pathogenesis of the disease. This review aimed to assess the differences in gene expression patterns and NAFLD/NASH progression pathways among the most common dietary animal models, i.e., HFD- and MCD diet-fed animals. Studies showed that the HFD and MCD diet could induce either up- or downregulation of the expression of genes and proteins that are involved in lipid metabolism, inflammation, oxidative stress, and fibrogenesis pathways. Interestingly, the MCD diet model could spontaneously develop liver fibrosis within two to four weeks and has significant effects on the expression of genes that encode proteins and enzymes involved in the liver fibrogenesis pathway. However, such effects in the HFD model were found to occur after 24 weeks with insulin resistance but appear to cause less severe fibrosis. In conclusion, assessing the abnormal gene expression patterns caused by different diet types provides valuable information regarding the molecular mechanisms of NAFLD/NASH and predicts the clinical progression of the disease. However, expression profiling studies concerning genetic variants involved in the development and progression of NAFLD/NASH should be conducted

Hepatoprotective, immunomodulatory and antibacterial effects of selected Malaysian medicinal plant extracts / Mohammed Abdullah Mahdi Alshawsh

Orthosiphon stamineus Benth and Morinda citrifolia L. are considered an important traditional folk medicine and commonly used in Malaysia for treating many diseases. In this study, the ethanol extracts of O. stamineus and M. citrifolia were evaluated for their hepatoprotective and antioxidant activities; in vivo against thioacetamide-induced liver cirrhosis in rats and in vitro against H2O2-induced hepatotoxicity in WRL-68 liver cell line, as well as to investigate their immunomodulatory and antibacterial effects. Orthosiphon stamineus and Morinda citrifolia were evaluated for their antioxidant activities using DPPH, ABTS and FRAP. In addition the phenolic and flavonoids contents were determined. Seven groups of adult SD rats were used in the hepatoprotective experiment. Group 1 as normal control group, while groups 2 to 7 were injected intraperitoneally with 200 mg/kg of thioacetamide (TAA) thrice weekly for two months and orally administered respectively with 10 % Tween 20, 50 mg/kg silymarin, 200 mg/kg and 100 mg/kg ethanol extract of O. stamineus and M. citrifolia daily for two months. The hepatoprotective activity was evaluated using the following parameters; body and liver weight, serum liver biochemical markers, liver gross morphology and histopathology, as well as endogenous antioxidant markers. Furthermore, the liver fibrosis related genes namely; TGFβ1, MMP2, TIMP1 and Coll α were estimated for the change in gene expression levels using RT-PCR. In addition, the ethanol crude extracts of O. stamineus and M. citrifolia and their isolated fractions were investigated against H2O2-induced hepatotoxicity in WRL-68 liver cell line and the percentage of cell viability using MTT assay and the antioxidant level markers were assessed. The immunomodulatory potential of the extracts were investigated by MTT assay against human peripheral blood mononuclear cells (PBMCs), while the antibacterial activity was investigated by disc diffusion and determination of minimum iii inhibitory concentration (MIC) against four Gram-positive and Gram-negative bacterial strains. Finally, LC-MS was used for identification of the active constituents of the fractions that proved to have hepatoprotective activity. Orthosiphon stamineus exhibited significant free radical scavenging activity with DPPH (IC50 21.4 μg/ml), at the same time, showed high total phenolic and flavonoidal contents. In animal experiments, the hepatotoxic group showed a coarse granulation on the liver surface when compared to the smooth aspect observed on the liver surface of normal and treatment groups. Histopathological study confirmed the result. Moreover, there was a significant (P < 0.05) increase in serum liver biochemical parameters (ALT, AST, ALP and bilirubin) and the level of liver lipid peroxidation index malondialdehyde (MDA), accompanied by a significant decrease in the level of total protein, albumin, catalase, superoxide dismutase and glutathione peroxidase in the TAA control group comparing with normal group. The 200 mg/kg treatment groups of both plants significantly restored the elevated liver function enzymes and antioxidant parameters near to normal and significantly down-regulated the expression of the liver fibrosis genes. The oxidative stress by H2O2 resulted in a decrease of cell viability to 41.9 %, while pre-treatment with crude extracts of O. stamineus and M. citrifolia, as well as with fraction 3 of O. stamineus and fraction 2 of M. citrifolia were found significantly (P < 0.01) increase the cell viability to 81.1 %, 76.4 %, 95.1 % and 86.1 % respectively at concentration 100 μg/ml. The hepatoprotective and antioxidant activity could be claimed to the following flavonoids; ponkanetin, eupatorin, TMF and salvigenin that were identified in O. stamineus F3 and scopoletin and P- coumaric acid, which were identified in M. citrifolia F2. In conclusion, this study showed that O. stamineus and M. citrifolia exhibit potent antioxidant properties, immunomodulatory activity and could be an effective herbal and efficient remedy for chemical-induced hepatic cirrhosis, thus may be a highly promising candidate drugs

Diosmetin Exerts Synergistic Effects in Combination with 5-Fluorouracil in Colorectal Cancer Cells

5-Fluorouracil (5-FU) is a chemotherapeutic medication commonly used to treat colorectal cancer (CRC); however, the drug-associated adverse effects and toxicity have greatly affected its clinical use. Exploring another therapeutic strategy that lowers the toxicity of 5-FU while having a synergistic effect against CRC is thus a viable option. Diosmetin, a natural flavonoid, has been shown to inhibit the proliferation of many cancer cells, including CRC cells. This study aims to investigate the synergistic effect of diosmetin and 5-FU on HCT116 and HT29 colorectal cancer cells and to explore the apoptotic activity of this combination. The MTT assay was used to assess the viability of cells treated with monotherapy and combination therapy. The combination index (CI) and dose reduction index (DRI) were calculated using the CompuSyn software (version 1.0). The SynergyFinder 2.0 software was used to calculate the synergy score, while the Combenefit software was employed to perform isobologram analysis and synergism determination. The AO/PI double staining technique was used to detect the apoptotic characteristics of cells, whereas the flow cytometry technique was used to investigate the apoptosis induction and cell cycle arrest in cells. The combination of 5-FU and diosmetin showed a synergistic effect in HCT116 cells with a mean CI value of 0.66 &plusmn; 0.4, and an additive effect in HT29 cells with a CI value of 1.0 &plusmn; 0.2. The DRI of 5-FU in HCT116 cells was three times lower in the combination therapy compared to monotherapy of 5-FU. AO/PI microscopic examination and Annexin V analysis revealed that the combination-treated cells had more apoptotic cells than the monotherapy-treated cells, which was activated mainly through intrinsic apoptosis pathway. HCT116 cell death was confirmed by mitotic arrest in the G2/M phase. Our findings suggest that 5-FU/diosmetin combination exhibits synergistic effect against HCT116 cancer cells, and potentially reduces the unfavorable adverse effect of 5-FU while enhancing the anticancer efficacy by inducing apoptosis and interrupting mitosis. Further research studies are needed to validate the combination&rsquo;s anti-tumorigenic activities in a xenograft animal model

<i>Faecalibacterium prausnitzii</i> Ameliorates Colorectal Tumorigenesis and Suppresses Proliferation of HCT116 Colorectal Cancer Cells

Faecalibacterium prausnitzii is one of the most abundant commensals of gut microbiota that is not commonly administered as a probiotic supplement. Being one of the gut’s major butyrate-producing bacteria, its clinical significance and uses are on the rise and it has been shown to have anti-inflammatory and gut microbiota-modulating properties in the treatment of inflammatory bowel illness, Crohn’s disease, and colorectal cancer. Colorectal cancer (CRC) is a silent killer disease that has become one of the leading causes of cancer-related death worldwide. This study aimed to evaluate the anti-tumorigenic and antiproliferative role of F. prausnitzii as well as to study its effects on the diversity of gut microbiota in rats. Findings showed that F. prausnitzii probiotic significantly reduced the colonic aberrant crypt foci frequency and formation in Azoxymethane (AOM)-induced CRC in rats. In addition, the administration of F. prausnitzii lowered the lipid peroxidation levels in the colon tissues. For in vitro 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay, the cell-free supernatant of F. prausnitzii suppressed the growth of HCT116 colorectal cancer cells in a time/dose-dependent manner. 16S rRNA gene sequencing using rat stool samples showed that the administration of F. prausnitzii modulated the gut microbiota of the rats and enhanced its diversity. Hence, these findings suggest that F. prausnitzii as a probiotic supplement can be used in CRC prevention and management; however, more studies are warranted to understand its cellular and molecular mechanisms of action

Therapeutic Potential of Certain Terpenoids as Anticancer Agents: A Scoping Review

Cancer is a life-threatening disease and is considered to be among the leading causes of death worldwide. Chemoresistance, severe toxicity, relapse and metastasis are the major obstacles in cancer therapy. Therefore, introducing new therapeutic agents for cancer remains a priority to increase the range of effective treatments. Terpenoids, a large group of secondary metabolites, are derived from plant sources and are composed of several isoprene units. The high diversity of terpenoids has drawn attention to their potential anticancer and pharmacological activities. Some terpenoids exhibit an anticancer effect by triggering various stages of cancer progression, for example, suppressing the early stage of tumorigenesis via induction of cell cycle arrest, inhibiting cancer cell differentiation and activating apoptosis. At the late stage of cancer development, certain terpenoids are able to inhibit angiogenesis and metastasis via modulation of different intracellular signaling pathways. Significant progress in the identification of the mechanism of action and signaling pathways through which terpenoids exert their anticancer effects has been highlighted. Hence, in this review, the anticancer activities of twenty-five terpenoids are discussed in detail. In addition, this review provides insights on the current clinical trials and future directions towards the development of certain terpenoids as potential anticancer agents

Medicinal Plants and Their Inhibitory Activities against Pancreatic Lipase: A Review

Obesity is recognized as a major life style disorder especially in developing countries and it is prevailing at an alarming speed in new world countries due to fast food intake, industrialization, and reduction of physical activity. Furthermore, it is associated with a vast number of chronic diseases and disabilities. To date, relatively effective drugs, from either natural or synthetic sources, are generally associated with serious side effects, often leading to cessation of clinical trials or even withdrawal from the market. In order to find new compounds which are more effective or with less adverse effects compared to orlistat, the drug that has been approved for obesity, new compounds isolated from natural products are being identified and screened for antiobesity effects, in particular, for their pancreatic lipase inhibitory effect. Pancreatic lipase inhibitory activity has been extensively used for the determination of potential efficacy of natural products as antiobesity agents. In attempts to identify natural products for overcoming obesity, more researches have been focused on the identification of newer pancreatic lipase inhibitors with less unpleasant adverse effects. In this review, we consider the potential role of plants that have been investigated for their pancreatic lipase inhibitory activity

Antibacterial Mechanism of Action of Two Types of Honey against Escherichia coli through Interfering with Bacterial Membrane Permeability, Inhibiting Proteins, and Inducing Bacterial DNA Damage

Honey is a sweet natural food produced by bees from flower nectar or some part of plant secretions that exhibit antimicrobial activity against many microorganisms. It has been used as traditional therapy for skin infections. Antibiotics play an essential role in managing wound infection; however, some pathogenic bacteria have begun to possess resistance against them, which may cause chronic infections and severe adverse effects. This study investigates the antibacterial activities and mechanism of action of Yemeni Sidr honey (SH) and Manuka honey (MH) against Escherichia coli. The inhibitory effects of SH and MH using the disk diffusion method on bacterial growth were remarkable at 700 mg/disk. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were similar for both kinds of honey. However, MH showed a better bactericidal effect (30%) than SH (50%). The antimicrobial mechanism of action showed that SH substantially impacted the bacterial membrane&rsquo;s permeability and increased the potassium and protein leakage rate. On the contrary, MH demonstrated remarkable inhibition of bacterial protein synthesis, while both kinds of honey caused bacterial DNA damage. These data reveal that SH and MH could be used as a remedy for skin infections and might be further developed as a promising dressing for bacterial wound infections

Cynometra cauliflora Linn. Attenuates metabolic abnormalities in high-fat diet-induced obese mice

Ethnopharmacological relevance: Cynometra cauliflora Linn. belongs to the Fabaceae family and is known locally in Malaysia as nam-nam. Traditionally, a decoction of the C. cauliflora leaves is used for treating hyperlipidemia and diabetes. Aim of the study: This study aims to investigate the anti-obesity and lipid lowering effects of ethanolic extract of C. cauliflora leaves and its major compound (vitexin) in C57BL/6 obese mice induced by high-fat diet (HFD), as well as to further identify the molecular mechanism underlying this action. Methods and material: Male C57BL/6 mice were fed with HFD (60% fat) for 16 weeks to become obese. The treatment started during the last 8 weeks of HFD feeding and the obese mice were treated with C. cauliflora leaf extract at 200 and 400 mg/kg/day, orlistat (10 mg/kg) and vitexin (10 mg/kg). Results: The oral administration of C. cauliflora (400 and 200 mg/kg) and vitexin significantly reduced body weight, adipose tissue and liver weight and lipid accumulation in the liver compared to control HFD group. Both doses of C. cauliflora also significantly (P ≤ 0.05) decreased serum triglyceride, LDL, lipase, IL-6, peptide YY, resistin levels, hyperglycemia, hyperinsulinemia, and hyperleptinemia compared to the control HFD group. Moreover, C. cauliflora significantly up-regulated the expression of adiponectin, Glut4, Mtor, IRS-1 and InsR genes, and significantly decreased the expression of Lepr in white adipose tissue. Furthermore, C. cauliflora significantly up-regulated the expression of hypothalamus Glut4, Mtor and NF-kB genes. GC-MS analysis of C. cauliflora leaves detected the presence of phytol, vitamin E and β-sitosterol. Besides, the phytochemical evaluation of C. cauliflora leaves showed the presence of flavonoid, saponin and phenolic compounds. Conclusion: This study shows interesting outcomes of C. cauliflora against HFD-induced obesity and associated metabolic abnormalities. Therefore, the C. cauliflora extract could be a potentially effective agent for obesity management and its related metabolic disorders such as insulin resistance and hyperlipidemia. © 2019 Elsevier B.V

Investigation into the Potential Role of <i>Propionibacterium freudenreichii</i> in Prevention of Colorectal Cancer and Its Effects on the Diversity of Gut Microbiota in Rats

Colorectal cancer (CRC) accounts for 10% of all cancer diagnoses and cancer-related deaths worldwide. Over the past two decades, several studies have demonstrated the clinical benefits of probiotic supplementation and some studies have shown that certain probiotics can modulate immunity and strengthen gut microbiota diversity. This study aims to assess the impact of the Propionibacterium freudenreichii (PF) probiotic against CRC induced by azoxymethane (AOM), and to investigate its effects on gut microbiota diversity in rats, as well as to evaluate the anti-proliferative activities of PF in HCT116 CRC cells. This experiment was performed using four groups of SD rats: normal control, AOM group, PF group (1 × 109 CFU/mL), and standard drug control (5-fluorouracil, 35 mg/kg). Methylene blue staining of colon tissues showed that the administration of PF significantly reduced the formation of colonic aberrant crypt foci (ACF) compared to the AOM control group. In addition, treated rats had lower levels of malondialdehyde in their colon tissue homogenates, indicating that lipid peroxidation was suppressed by PF supplementation. Furthermore, 16S rRNA gene analysis revealed that probiotic treatment enhanced the diversity of gut microbiota in rats. In vitro study showed that the viability of HCT116 cells was inhibited by the probiotic cell-free supernatant with an IC50 value of 13.3 ± 0.133. In conclusion, these results reveal that consuming PF as probiotic supplements modulates gut microbiota, inhibits the carcinogenic effects of AOM, and exerts anti-proliferative activity against CRC cells. Further studies are required to elucidate the role of PF on the immune response during the development and growth of CRC