Pub Date : 2019-08-28DOI: 10.24870/cjb.2019-000129
R. Sunday, E. Obuotor, Anil Kumar
Background: The antidiabetic effect of Asparagus adscendens root ethanolic extract (AAE) was evaluated in this study using both in vivo and in vitro models. The effect of AAE on carbohydrate metabolizing enzymes (α-amylase and α-glucosidase) was determined. The safety of AAE was tested on Wistar rats and two different cell lines. Some mechanisms of action were also investigated with AAE’s dose-response. Methods: Glucose-loaded (10 g/kg) and streptozotocin-induced (60 mg/kg) diabetic Wistar rats were used in the in vivo model, whereas RIN-5F pancreatic cells and HepG2 liver cells were used in the in vitro model. Nontoxic mass value of AAE was used in the in vitro (from 0.625 to 2.5 μg/100 µl) and in vivo (up to 400 mg/kg) studies. The inhibitory activity of AAE on α-amylase and α-glucosidase was examined by spectrophotometric and microplate reader techniques. Results: The AAE inhibited α-amylase and α-glucosidase, two key enzymes of the carbohydrate metabolism, and stimulated the release of insulin in RIN-5F cells line and glucose uptake in HepG2 cells in a concomitant way. Lower mass values of the extract caused no significant change in the viability of the cells, whereas 5 μg caused a significant reduction in the viability of RIN-5F (59.78%) and HepG2 (56.87%) when compared to the control. The 2.5 μg extract stimulated 91% insulin release in RIN-5F cells when compared with the control. Also, 2.5 μg extract induced 86% and 83% glucose uptake in HepG2 cells in the presence and absence of insulin, respectively, when compared with the control. The median lethal dose of AAE was ≥5000 mg/kg in Wistar rats. AAE caused a decrease in fasting blood glucose level from 30 min in glucose-loaded Wistar rats and from the 4 th day in streptozotocin-induced diabetic rats when compared with the control. There was also an increase in serum insulin and serum α-amylase level in streptozotocin-induced diabetic rats, compared to the control, at the end of the study. Conclusion: A. adscendens root exerts its antidiabetic effect by inhibiting α-amylase and α-glucosidase enzymes, inducing insulin secretion in RIN-5F pancreatic cells, and enhancing glucose uptake in HepG2 liver cells.
{"title":"Antidiabetic Effect of Asparagus adscendens Roxb. in RIN-5F Cells, HepG2 Cells, and Wistar Rats","authors":"R. Sunday, E. Obuotor, Anil Kumar","doi":"10.24870/cjb.2019-000129","DOIUrl":"https://doi.org/10.24870/cjb.2019-000129","url":null,"abstract":"Background: The antidiabetic effect of Asparagus adscendens root ethanolic extract (AAE) was evaluated in this study using both in vivo and in vitro models. The effect of AAE on carbohydrate metabolizing enzymes (α-amylase and α-glucosidase) was determined. The safety of AAE was tested on Wistar rats and two different cell lines. Some mechanisms of action were also investigated with AAE’s dose-response. Methods: Glucose-loaded (10 g/kg) and streptozotocin-induced (60 mg/kg) diabetic Wistar rats were used in the in vivo model, whereas RIN-5F pancreatic cells and HepG2 liver cells were used in the in vitro model. Nontoxic mass value of AAE was used in the in vitro (from 0.625 to 2.5 μg/100 µl) and in vivo (up to 400 mg/kg) studies. The inhibitory activity of AAE on α-amylase and α-glucosidase was examined by spectrophotometric and microplate reader techniques. Results: The AAE inhibited α-amylase and α-glucosidase, two key enzymes of the carbohydrate metabolism, and stimulated the release of insulin in RIN-5F cells line and glucose uptake in HepG2 cells in a concomitant way. Lower mass values of the extract caused no significant change in the viability of the cells, whereas 5 μg caused a significant reduction in the viability of RIN-5F (59.78%) and HepG2 (56.87%) when compared to the control. The 2.5 μg extract stimulated 91% insulin release in RIN-5F cells when compared with the control. Also, 2.5 μg extract induced 86% and 83% glucose uptake in HepG2 cells in the presence and absence of insulin, respectively, when compared with the control. The median lethal dose of AAE was ≥5000 mg/kg in Wistar rats. AAE caused a decrease in fasting blood glucose level from 30 min in glucose-loaded Wistar rats and from the 4 th day in streptozotocin-induced diabetic rats when compared with the control. There was also an increase in serum insulin and serum α-amylase level in streptozotocin-induced diabetic rats, compared to the control, at the end of the study. Conclusion: A. adscendens root exerts its antidiabetic effect by inhibiting α-amylase and α-glucosidase enzymes, inducing insulin secretion in RIN-5F pancreatic cells, and enhancing glucose uptake in HepG2 liver cells.","PeriodicalId":166744,"journal":{"name":"Canadian Journal of Biotechnology","volume":"57 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2019-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"128340368","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-04-28DOI: 10.24870/CJB.2019-000126
O. Ovuakporie-Uvo, M. Idu, Omotuyi I. Olaposi
Background: Toxicity studies are relevant in assessing the side effects of chemical substances before they are incorporated into the process of drug development. Introduction: Desplatsia dewevrei is a scarce forest species believed by natives to be nutritive and therapeutic, without scientific evidence though. Thus, this study was aimed at investigating the possible toxicity of shortand long-term oral administration of D. dewevrei using Wistar rats. Methods: 0, 30 100, and 1000 mg/kg of D. dewevrei were daily administered p.o for 3 and 28 days to Wistar rats consisting of four animals (two females, two males) per group. Hemotoxicity and liver function tests were done using automated machines from ERMA Inc. RT-PCR method was used to study the regulation of intestinal glucose transporter 4 (GLUT4), glucose transporter 2 (GLUT2), glucagon-like peptide-1 (GLP-1), pancreatic insulin, KCJN5, and L-type voltage-gated calcium channel genes (CACNAIA). Results: No morphological or hematological signs of toxicity were observed. Liver function test showed an elevated level of highdensity lipoprotein (HDL-C) in the treatment group (100 mg/kg). The lethal dose (LD50) of D. dewevrei extracts were above 1000 mg/kg as no mortality was observed at the highest regimen dose used. Up-regulation of pancreatic insulin and down-regulation of intestinal GLUT-2 suggest that the plant may contain therapeutic constituents. Conclusion: Shortor long-term administration of D. dewevrei is relatively safe.
{"title":"Toxicogenetic Studies of Desplatsia dewevrei using Gene Expression of Blood, Pancreatic, and Intestinal Genes in Wistar rats","authors":"O. Ovuakporie-Uvo, M. Idu, Omotuyi I. Olaposi","doi":"10.24870/CJB.2019-000126","DOIUrl":"https://doi.org/10.24870/CJB.2019-000126","url":null,"abstract":"Background: Toxicity studies are relevant in assessing the side effects of chemical substances before they are incorporated into the process of drug development. Introduction: Desplatsia dewevrei is a scarce forest species believed by natives to be nutritive and therapeutic, without scientific evidence though. Thus, this study was aimed at investigating the possible toxicity of shortand long-term oral administration of D. dewevrei using Wistar rats. Methods: 0, 30 100, and 1000 mg/kg of D. dewevrei were daily administered p.o for 3 and 28 days to Wistar rats consisting of four animals (two females, two males) per group. Hemotoxicity and liver function tests were done using automated machines from ERMA Inc. RT-PCR method was used to study the regulation of intestinal glucose transporter 4 (GLUT4), glucose transporter 2 (GLUT2), glucagon-like peptide-1 (GLP-1), pancreatic insulin, KCJN5, and L-type voltage-gated calcium channel genes (CACNAIA). Results: No morphological or hematological signs of toxicity were observed. Liver function test showed an elevated level of highdensity lipoprotein (HDL-C) in the treatment group (100 mg/kg). The lethal dose (LD50) of D. dewevrei extracts were above 1000 mg/kg as no mortality was observed at the highest regimen dose used. Up-regulation of pancreatic insulin and down-regulation of intestinal GLUT-2 suggest that the plant may contain therapeutic constituents. Conclusion: Shortor long-term administration of D. dewevrei is relatively safe.","PeriodicalId":166744,"journal":{"name":"Canadian Journal of Biotechnology","volume":"1 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2019-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"129870586","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2018-11-28DOI: 10.24870/CJB.2018-000123
J. Maixent, M. Fares, C. François
Background: Herbal plant extracts are a more common alternative to conventional medicine to treat sleep disorders and intermittent hypoxia. Notably, obstructive sleep apnea causes injuries similar to those observed in models of ischemia-reperfusion including the decrease of nitric oxide (NO) availability. Kidney transplantation in end-stage renal disease reverses the sleep apnea. The underlying mechanism linking hypoxia, sleep apnea, and renal protection remains to be defined at the cellular level. Objective: The aim of this study was to demonstrate the safety and efficacy of herbal plant infusions with a potential for donating NO, to attenuation of damage induced during a hypoxia/reperfusion sequence, on kidney epithelial cells LLC-PK1. Materials and Methods: Cell death (Lactate Dehydrogenase release assay) and a viability test (MTS assay) after 24 h of incubation with different concentrations of plant infusion were assessed using the LLC-PK1 cell line. Then, measurement of the breakdown product of NO (the NaNO2) and LDH assay were carried out after 24 h of hypoxia, followed by 4 h or 24 h of reperfusion. Results: The effect of different dilutions of herbal plant infusion on the LLC-PK1 cell viability, after 24 h of incubation, was maximal at a 30% dilution compared to control. After 24 h of hypoxia, there was an increase of NaNO2 and thus of NO, and a concentration-dependent decrease of cell death. Similar results were observed after hypoxia followed by 4 h of reperfusion. These effects were always maximal at 50% dilution of plants infusion. Conclusion: Safe infusion of plant extracts causes a dose-dependent increase of NO and has a protective effect against the cellular stress caused by hypoxia and reoxygenation. Since it has been demonstrated that there is a NO-dependent mechanism allowing the reduction of injuries induced by ischemia/reoxygenation process, such a mechanism could be responsible for our observations.
{"title":"Infusion of herbal plant extracts for insomnia and anxiety causes a dose-dependent increase of NO and has a protective effect on the renal cellular stress caused by hypoxia and reoxygenation","authors":"J. Maixent, M. Fares, C. François","doi":"10.24870/CJB.2018-000123","DOIUrl":"https://doi.org/10.24870/CJB.2018-000123","url":null,"abstract":"Background: Herbal plant extracts are a more common alternative to conventional medicine to treat sleep disorders and intermittent hypoxia. Notably, obstructive sleep apnea causes injuries similar to those observed in models of ischemia-reperfusion including the decrease of nitric oxide (NO) availability. Kidney transplantation in end-stage renal disease reverses the sleep apnea. The underlying mechanism linking hypoxia, sleep apnea, and renal protection remains to be defined at the cellular level. Objective: The aim of this study was to demonstrate the safety and efficacy of herbal plant infusions with a potential for donating NO, to attenuation of damage induced during a hypoxia/reperfusion sequence, on kidney epithelial cells LLC-PK1. Materials and Methods: Cell death (Lactate Dehydrogenase release assay) and a viability test (MTS assay) after 24 h of incubation with different concentrations of plant infusion were assessed using the LLC-PK1 cell line. Then, measurement of the breakdown product of NO (the NaNO2) and LDH assay were carried out after 24 h of hypoxia, followed by 4 h or 24 h of reperfusion. Results: The effect of different dilutions of herbal plant infusion on the LLC-PK1 cell viability, after 24 h of incubation, was maximal at a 30% dilution compared to control. After 24 h of hypoxia, there was an increase of NaNO2 and thus of NO, and a concentration-dependent decrease of cell death. Similar results were observed after hypoxia followed by 4 h of reperfusion. These effects were always maximal at 50% dilution of plants infusion. Conclusion: Safe infusion of plant extracts causes a dose-dependent increase of NO and has a protective effect against the cellular stress caused by hypoxia and reoxygenation. Since it has been demonstrated that there is a NO-dependent mechanism allowing the reduction of injuries induced by ischemia/reoxygenation process, such a mechanism could be responsible for our observations.","PeriodicalId":166744,"journal":{"name":"Canadian Journal of Biotechnology","volume":"47 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2018-11-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"127707902","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2018-09-28DOI: 10.24870/CJB.2018-000122
R. Gómez-García, M. A. Medina-Morales, R. Rodríguez, B. Farruggia, G. Picó, Cristóbal N. Aguilar
Production of xylanase enzyme by fungi strains has gained interest in the recent years due to its high productivity, high catalytic power, as well as its potential applications in different areas such as feed, food, textile, and biofuel industries. The conventional methodologies, to produce enzymes, involve complex apparatus and chemical solvents and are associated with high costs and lowyield recovery. To obtain the high-yield recovery of the enzymes, modern enzyme extraction methods are taken into account. Aqueous two-phase systems (ATPS) are an alternative separative methodology for the purification and recovery of the enzymes and other biomolecules. The advantages of ATPS are easy scale-up and extraction, volume reduction, and rapid separation. The objective of this study was to produce Trichoderma harzianum xylanase by solid-state fermentation (SSF) using corn cobs as a support/substrate and employing ATPS for its partial recovery. In this study, the results showed the ability of a microorganism to grow on the corn cobs and to produce the xylanase enzyme. Xylanolytic activity reached 7.85 U/g of corn cobs. The enzyme was efficiently concentrated by ATPS. In addition, a high purification factor (10-fold) and considerable enzyme recovery (%ER) (84%) percentage were obtained.
{"title":"Production of a xylanase by Trichoderma harzianum (Hypocrea lixii) in solid-state fermentation and its recovery by an aqueous two-phase system","authors":"R. Gómez-García, M. A. Medina-Morales, R. Rodríguez, B. Farruggia, G. Picó, Cristóbal N. Aguilar","doi":"10.24870/CJB.2018-000122","DOIUrl":"https://doi.org/10.24870/CJB.2018-000122","url":null,"abstract":"Production of xylanase enzyme by fungi strains has gained interest in the recent years due to its high productivity, high catalytic power, as well as its potential applications in different areas such as feed, food, textile, and biofuel industries. The conventional methodologies, to produce enzymes, involve complex apparatus and chemical solvents and are associated with high costs and lowyield recovery. To obtain the high-yield recovery of the enzymes, modern enzyme extraction methods are taken into account. Aqueous two-phase systems (ATPS) are an alternative separative methodology for the purification and recovery of the enzymes and other biomolecules. The advantages of ATPS are easy scale-up and extraction, volume reduction, and rapid separation. The objective of this study was to produce Trichoderma harzianum xylanase by solid-state fermentation (SSF) using corn cobs as a support/substrate and employing ATPS for its partial recovery. In this study, the results showed the ability of a microorganism to grow on the corn cobs and to produce the xylanase enzyme. Xylanolytic activity reached 7.85 U/g of corn cobs. The enzyme was efficiently concentrated by ATPS. In addition, a high purification factor (10-fold) and considerable enzyme recovery (%ER) (84%) percentage were obtained.","PeriodicalId":166744,"journal":{"name":"Canadian Journal of Biotechnology","volume":"77 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2018-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134057260","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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