Guava (Psidium guajava Linn.) leaf extract promotes glucose uptake and glycogen accumulation by modulating the insulin signaling pathway in high-glucose-induced insulin-resistant mouse FL83B cells

doi:10.1016/j.procbio.2015.03.022

Process Biochemistry

Volume 50, Issue 7, July 2015, Pages 1128-1135

https://doi.org/10.1016/j.procbio.2015.03.022 Get rights and content

Highlights

•
We reveal the hypoglycemic mechanisms of GvEx in high-glucose-induced insulin-resistant liver cells.
•
Insulin-resistant status was improved by GvEx through modulating the insulin signaling cascade.
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GvEx increases glucose uptake and enhanced glycogen accumulation.

Abstract

The effects of aqueous guava leaf extract (GvEx) on insulin resistance were evaluated in high glucose-induced insulin-resistant mouse FL83B cells. Glucose uptake and glycogen content were determined and western blot analysis was performed to study the anti-diabetic effects of GvEx. Our results showed that administration of 200 and 400 μg/mL GvEx resulted in a significant increase in glucose uptake in both normal and insulin-resistant cells. Glycogen content in cells was significantly higher after incubation with GvEx (200, 400 μg/mL) than that in control group (p < 0.05). Further, western blot analysis revealed that 200 and 400 μg/mL GvEx significantly enhanced the levels of insulin-related signaling components, including insulin receptor (IR), as well as enhancing the phosphorylation of the insulin receptor (p-IR (Tyr)), insulin receptor substrate (p-IRS (Tyr)), p85 regulatory subunit of phospho-inositide 3 kinase (PI3K (p85)), protein kinase B (p-Akt (Ser)), glucose transporter 2 (Glut 2), and total glycogen synthase. In conclusion, GvEx improved insulin resistance by promoting glucose uptake and enhancing glycogen content via modulation of the insulin signaling pathway in high glucose-induced insulin-resistant mouse FL83B cells.

Graphical abstract

Introduction

Diabetes mellitus (DM) is one of the most prevalent metabolic disorders and has reached epidemic proportions. It is estimated that 552 million will be diagnosed with DM by the year 2030 [1]. DM is characterized by hyperglycemia, associated with deficiencies in insulin secretion and/or insulin action. In particular, 90–95% cases of diagnosed diabetes are non-insulin-dependent, which is largely associated with diet, behavior, and excess body weight [2], [3]. Insulin is a major anabolic hormone that regulates glucose homeostasis and stimulates glucose transport. Insulin resistance is recognized as an important risk for the development of non-insulin-dependent or type 2 diabetes [4]. Therefore, identifying novel ways to improve of insulin resistance and insulin sensitivity is of great importance.

Psidium guajava, a member of the Myrtaceae family, is one of the most important economic fruits in tropical areas. Guava leaf has traditionally been used as a medicinal herb for diabetes patients in Oriental countries [5], [6], and is reported to contain many bioactive compounds, including tannins, polyphenols, flavonoids, triterpenoids, and quercetin, which could regulate hyperglycemia [7]. Indeed, guava leaf extracts can improve glucose metabolism and insulin resistance in the skeletal muscles of diabetic rats by modulating insulin-related signaling. The IRS-1, Akt, PI3K (p85) protein expression, then IRS-1, AMP-activated protein kinase, Akt (Thr 308) phosphorylation were promoted by guava leaf extracts. The enhanced insulin-related signaling should induce Glut 4 translocation. The ratios of membrane to total Glut 4 expression were observed increase in the skeletal muscles [8]. Additionally, guava leaf extract and/or its formula can improve insulin resistance by regulating the insulin signal transduction pathway and promoting glucose uptake in 3T3-L1 adipocytes. The expression of IR mRNA, and the protein expression and phosphorylation of PI3K (p85) and Akt (Ser473) were up-regulated. These induced insulin signal transduction pathways can further promote the Glut 4 mRNA expression and glucose uptake [6], [9]. The effects of guava leaf extracts on hyperglycemia in hepatocytes has been studied, and data suggest that it may promote glucose uptake in rat clone 9 hepatocytes and increase the activity of hepatic hexokinase and glucose-6-phosphate dehydrogenase in the liver of STZ-induced diabetic SD rats [10], [11]. However, the mechanism by which guava leaf extracts improves insulin resistance and modulates insulin-related signaling in hepatocytes remains unclear.

Declined insulin-stimulated tyrosine phosphorylation of insulin receptor substrate (p-IRS) by high glucose reduces insulin-stimulated phosphorylation of Akt, thereby inhibiting the metabolic effects of glucose uptake [12]. In the present study, FL83B cells from mouse liver were treated with high glucose to induce insulin resistance to evaluate the anti-hyperglycemic effects of aqueous guava leaf extract (GvEx). The glucose uptake and glycogen content in insulin-resistant FL83B cells were monitored. In addition, the expression of insulin signal-related proteins was also studied to elucidate the anti-hyperglycemic mechanism of GvEx in FL83B cells.

Section snippets

Plant material

The leaves of Jen Ju Pa (Psidium Guajava Linn.) were collected from Jing-cin Farm (Tianzhong Township, Changhua County, Taiwan) between the initial appearance and the visible opening of flower buds. The plant materials were taxonomically identified and deposited in the Fengshan Tropical Horticultural Experiment Branch, Taiwan Agricultural Research Institute Council of Agriculture, Executive Yuan (FTHA000282).

GvEx preparation

Ultrasound assisted optimal extraction conditions, which were derived from a response

Total phenolic content and chromatographic profile of GvEx

GvEx was prepared using the ultrasound-assisted extraction conditions described in our previous study [13]. Total phenolic content of GvEx was 260.2 mg gallic acid equivalents/g extract (data not shown). This result was identical to our previous value of 261.2 mg gallic acid equivalents/g extract. In addition, the chromatographic profile contained 7 identifiable components, including gallic acid, chlorogenic acid, catechin, caffeic acid, epicatechin, epigallocatechin gallate (EGCG), and

Conclusions

In conclusion, we evaluated the effects of GvEx on glucose uptake and glycogen synthesis in high glucose-induced insulin-resistant FL83B cells. GvEx exhibits a good capacity to alleviate high glucose-induced insulin resistance by improving insulin signaling and promoting glucose uptake in FL83B cells. Therefore, our results may provide new insights into the mechanisms by which GvEx improves insulin resistance in the hepatocytes.

Acknowledgement

The authors are grateful for the financial support for this research by the Tunghai University of Taiwan, ROC, under the project of “Global Research and Education on Environment and Society (GREEnS)” and Grand no. GREEnS 4-3.

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The muscle quality of farmed fish has attracted widespread public attention. To understand a comprehensive molecular mechanism underlying the effects of artificial formula feed- (AF) and grass-feeding (GF) on muscle quality and growth of grass carp (Ctenopharyngodon idellus), muscular transcriptomics and proteomics of GF and AF C. idellus were characterized. A total of 834 DEGs (484 up-, 350 down-regulated genes) and 487 DEPs (271 up- and 216 down-regulations) were identified between groups. Then, the functional enrichment analysis of DEGs & DEPs (DEs) and the integration of multiple-Omics (SDFs), further reflected that the stronger myofiber differentiation and protein synthesis capacities are likely the main strategy of more muscle mass gain in AF C. idellus, while the muscle growth in GF C. idellus is consequent to the stronger muscle cell proliferation and hypertrophy. Secondly, there was a significant difference in muscle fiber types between GF and AF groups, this has a noticeable effect on muscle tenderness and quality. In combination with previously metabolic results, we established DEG-DEP-SDM interaction network, which validated that the slow-MyHC-I fiber, dense in AF C. idellus muscles, accumulated more glycogen and exhibited more powerful glycolytic activity. Meanwhile, the suppressed activity of eIF2α in AF C. idellus muscles promoted protein synthesis, the “Ubiquitin mediated proteolysis” was simultaneously restrained. However, the activities of TCA cycle and fatty acid oxidation were significantly enhanced in GF muscles (rich in fast-MyHC-IIb). Accordingly, the up-regulated ACSL1, ACADS, ppara, etc. were quantified in GF C. idellus samples with stronger fatty acids β-oxidation activity, then led to the reduction of cellular triglycerides and lipid droplet formation. Additionally, our results also provide key information for further understanding of the molecular mechanisms (e.g. MAPK and PI3K signaling pathways, Necroptosis and so on) of C. idellus under dietary treatments. In summary, the grass-feeding could effectively alleviate the negative effects of AF on fish, namely, reducing fat deposition, improving nutrients, flavor and texture of muscle, relieving insulin resistance and stress tolerance, finally lead to better nourishing and healthy C. idellus.
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The pathogenesis of diabetes can be linked to several signalling pathways, such as insulin signalling pathway (Huang and Chen, 2009; Liu et al., 2015; Vinayagam et al., 2018). Regulating the activity of enzymes in insulin, gluconeogenic and glycogenolytic pathways is one of the strategies for the treatment of diabetes mellitus (Liu et al., 2015). Glycogen phosphorylase and glycogen synthase are the two important regulatory enzymes that catalyse the rate-limiting steps of glycogen breakdown and synthesis, respectively.
Psidium guajava is a medicinal plant with antidiabetic properties and can be found growing in tropical and sub-tropical countries around the world.
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The study design involved preparation of an aqueous Psidium guajava leaf extract; investigation of its effect on muscle glycogen synthase and phosphorylase activities and possible protective effect on pancreas in streptozotocin induced diabetic male Sprague-Dawley rats and phytochemical study of the extract to identify further compounds with anti-diabetic potential.
Diabetes was induced in male Sprague-Dawley rats with a single intraperitoneal injection of 40 mg/kg streptozotocin. Normal and diabetic animals were treated with 400 mg/kg body weight of Psidium guajava leaves aqueous extract for a period of 14 days.
The treatment of diabetic animals with the Psidium guajava extract ameliorated damage to the pancreatic islets and enhanced lowering of blood glucose following a glucose load. Diabetes significantly decreased (P < 0.05) glycogen synthase activity by 31% compared to normal animals this being associated with a decrease in enzyme expression. Psidium guajava treatment returned the enzyme activity to near normal levels. In diabetic animals, glycogen phosphorylase activity was elevated but not significantly (P > 0.5) and treatment of both diabetic and normal animals with the extract reduced activity of the enzyme. The Psidium guajava extract also reduced glycogen phosphorylase expression in diabetic animals. Skeletal muscle glycogen content reduced by diabetes was increased as a result of treatment of diabetic animals with the Psidium guajava extract. Phytochemical analysis of the aqueous extract of Psidium guajava using Gas Chromatography-Mass Spectroscopy indicated the presence of triterpenes and phenolic compounds and we also report what we believe are three new compounds.
The results from this study suggest that the antidiabetic effects of Psidium guajava may be due to modulation of glycogen metabolism mediated by phenolic compounds and triterpenes present in the extract.
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Guava (Psidium guajava Linn.) leaf extract promotes glucose uptake and glycogen accumulation by modulating the insulin signaling pathway in high-glucose-induced insulin-resistant mouse FL83B cells

Highlights

Abstract

Graphical abstract

Introduction

Section snippets

Plant material

GvEx preparation

Total phenolic content and chromatographic profile of GvEx

Conclusions

Acknowledgement

Phytomedicine

Biochem Biophys Res Commun

Process Biochem

J Biol Chem

J Biol Chem

Anal Biochem

Food Chem

Phytother Res

J Ethnopharmacol

Trends Cell Biol

J Biol Chem

Food Chem Toxicol

Atherosclerosis

Am J Clin Nutr

Cancer Lett

IDF diabetes atlas

Antidiabetic activity of Terminalia pallida fruit in alloxan induced diabetic rats

J Ethnopharmacol

Diabetes epidemiology as a tool to trigger diabetes research and care

Diabetologia