Image_4_Antidiabetic, Antihyperlipidemic, Antioxidant, Anti-inflammatory Activities of Ethanolic Seed Extract of Annona reticulata L. in Streptozotocin Induced Diabetic Rats.JPEG

Annona reticulata L. (Bullock's heart) is a pantropic tree commonly known as custard apple, which is used therapeutically for a variety of maladies. The present research was carried out to evaluate the possible protective effects of Annona reticulata L. (A. reticulata) ethanolic seed extract on an experimentally induced type 2 diabetes rat model. Male Albino Wistar rats were randomly divided into five groups with six animals in each group viz., control rats in group I, diabetic rats in group II, diabetic rats with 50 and 100 mg/kg/bw of ethanolic seed extract of A. reticulata in groups III and IV, respectively, and diabetic rats with metformin in group V. Treatment was given for 42 consecutive days through oral route by oro-gastric gavage. Administration of A. reticulata seed extract to diabetes rats significantly restored the alterations in the levels of body weight, food and water intake, fasting blood glucose (FBG), insulin levels, insulin sensitivity, HbA1c, HOMA-IR, islet area and insulin positive cells. Furthermore, A. reticulata significantly decreased the levels of triglycerides, cholesterol, LDL, and significantly increased the HDL in diabetic rats. A. reticulata effectively ameliorated the enzymatic (ALT, AST, ALP, GGT) and modification of histopathological changes in diabetic rats. The serum levels of the BUN, creatinine levels, uric acid, urine volume, and urinary protein were significantly declined with a significant elevation in CCr in diabetic rats treated with A. reticulata. MDA and NO levels were significantly reduced with an enhancement in SOD, CAT, and GPx antioxidant enzyme activities in the kidney, liver, and pancreas of diabetic rats treated with A. reticulata. Diabetic rats treated with A. reticulata have shown up-regulation in mRNA expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2), NAD(P)H:quinone oxidoreductase 1 (NQO1), Heme oxygenase-1 (HO-1) and protein expression level of Nrf2 with diminution in Keap1 mRNA expression level in pancreas, kidney, and liver. From the outcome of the current results, it can be inferred that seed extract of A. reticulata exhibits a protective effect in diabetic rats through its anti-diabetic, anti-hyperlipidemic, antioxidant and anti-inflammatory effects and could be considered as a promising treatment therapy in the treatment of diabetes mellitus.</p

Image_2_Antidiabetic, Antihyperlipidemic, Antioxidant, Anti-inflammatory Activities of Ethanolic Seed Extract of Annona reticulata L. in Streptozotocin Induced Diabetic Rats.JPEG

Annona reticulata L. (Bullock's heart) is a pantropic tree commonly known as custard apple, which is used therapeutically for a variety of maladies. The present research was carried out to evaluate the possible protective effects of Annona reticulata L. (A. reticulata) ethanolic seed extract on an experimentally induced type 2 diabetes rat model. Male Albino Wistar rats were randomly divided into five groups with six animals in each group viz., control rats in group I, diabetic rats in group II, diabetic rats with 50 and 100 mg/kg/bw of ethanolic seed extract of A. reticulata in groups III and IV, respectively, and diabetic rats with metformin in group V. Treatment was given for 42 consecutive days through oral route by oro-gastric gavage. Administration of A. reticulata seed extract to diabetes rats significantly restored the alterations in the levels of body weight, food and water intake, fasting blood glucose (FBG), insulin levels, insulin sensitivity, HbA1c, HOMA-IR, islet area and insulin positive cells. Furthermore, A. reticulata significantly decreased the levels of triglycerides, cholesterol, LDL, and significantly increased the HDL in diabetic rats. A. reticulata effectively ameliorated the enzymatic (ALT, AST, ALP, GGT) and modification of histopathological changes in diabetic rats. The serum levels of the BUN, creatinine levels, uric acid, urine volume, and urinary protein were significantly declined with a significant elevation in CCr in diabetic rats treated with A. reticulata. MDA and NO levels were significantly reduced with an enhancement in SOD, CAT, and GPx antioxidant enzyme activities in the kidney, liver, and pancreas of diabetic rats treated with A. reticulata. Diabetic rats treated with A. reticulata have shown up-regulation in mRNA expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2), NAD(P)H:quinone oxidoreductase 1 (NQO1), Heme oxygenase-1 (HO-1) and protein expression level of Nrf2 with diminution in Keap1 mRNA expression level in pancreas, kidney, and liver. From the outcome of the current results, it can be inferred that seed extract of A. reticulata exhibits a protective effect in diabetic rats through its anti-diabetic, anti-hyperlipidemic, antioxidant and anti-inflammatory effects and could be considered as a promising treatment therapy in the treatment of diabetes mellitus.</p

Image_1_Antidiabetic, Antihyperlipidemic, Antioxidant, Anti-inflammatory Activities of Ethanolic Seed Extract of Annona reticulata L. in Streptozotocin Induced Diabetic Rats.TIF

Annona reticulata L. (Bullock's heart) is a pantropic tree commonly known as custard apple, which is used therapeutically for a variety of maladies. The present research was carried out to evaluate the possible protective effects of Annona reticulata L. (A. reticulata) ethanolic seed extract on an experimentally induced type 2 diabetes rat model. Male Albino Wistar rats were randomly divided into five groups with six animals in each group viz., control rats in group I, diabetic rats in group II, diabetic rats with 50 and 100 mg/kg/bw of ethanolic seed extract of A. reticulata in groups III and IV, respectively, and diabetic rats with metformin in group V. Treatment was given for 42 consecutive days through oral route by oro-gastric gavage. Administration of A. reticulata seed extract to diabetes rats significantly restored the alterations in the levels of body weight, food and water intake, fasting blood glucose (FBG), insulin levels, insulin sensitivity, HbA1c, HOMA-IR, islet area and insulin positive cells. Furthermore, A. reticulata significantly decreased the levels of triglycerides, cholesterol, LDL, and significantly increased the HDL in diabetic rats. A. reticulata effectively ameliorated the enzymatic (ALT, AST, ALP, GGT) and modification of histopathological changes in diabetic rats. The serum levels of the BUN, creatinine levels, uric acid, urine volume, and urinary protein were significantly declined with a significant elevation in CCr in diabetic rats treated with A. reticulata. MDA and NO levels were significantly reduced with an enhancement in SOD, CAT, and GPx antioxidant enzyme activities in the kidney, liver, and pancreas of diabetic rats treated with A. reticulata. Diabetic rats treated with A. reticulata have shown up-regulation in mRNA expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2), NAD(P)H:quinone oxidoreductase 1 (NQO1), Heme oxygenase-1 (HO-1) and protein expression level of Nrf2 with diminution in Keap1 mRNA expression level in pancreas, kidney, and liver. From the outcome of the current results, it can be inferred that seed extract of A. reticulata exhibits a protective effect in diabetic rats through its anti-diabetic, anti-hyperlipidemic, antioxidant and anti-inflammatory effects and could be considered as a promising treatment therapy in the treatment of diabetes mellitus.</p

Image_3_Antidiabetic, Antihyperlipidemic, Antioxidant, Anti-inflammatory Activities of Ethanolic Seed Extract of Annona reticulata L. in Streptozotocin Induced Diabetic Rats.JPEG

Annona reticulata L. (Bullock's heart) is a pantropic tree commonly known as custard apple, which is used therapeutically for a variety of maladies. The present research was carried out to evaluate the possible protective effects of Annona reticulata L. (A. reticulata) ethanolic seed extract on an experimentally induced type 2 diabetes rat model. Male Albino Wistar rats were randomly divided into five groups with six animals in each group viz., control rats in group I, diabetic rats in group II, diabetic rats with 50 and 100 mg/kg/bw of ethanolic seed extract of A. reticulata in groups III and IV, respectively, and diabetic rats with metformin in group V. Treatment was given for 42 consecutive days through oral route by oro-gastric gavage. Administration of A. reticulata seed extract to diabetes rats significantly restored the alterations in the levels of body weight, food and water intake, fasting blood glucose (FBG), insulin levels, insulin sensitivity, HbA1c, HOMA-IR, islet area and insulin positive cells. Furthermore, A. reticulata significantly decreased the levels of triglycerides, cholesterol, LDL, and significantly increased the HDL in diabetic rats. A. reticulata effectively ameliorated the enzymatic (ALT, AST, ALP, GGT) and modification of histopathological changes in diabetic rats. The serum levels of the BUN, creatinine levels, uric acid, urine volume, and urinary protein were significantly declined with a significant elevation in CCr in diabetic rats treated with A. reticulata. MDA and NO levels were significantly reduced with an enhancement in SOD, CAT, and GPx antioxidant enzyme activities in the kidney, liver, and pancreas of diabetic rats treated with A. reticulata. Diabetic rats treated with A. reticulata have shown up-regulation in mRNA expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2), NAD(P)H:quinone oxidoreductase 1 (NQO1), Heme oxygenase-1 (HO-1) and protein expression level of Nrf2 with diminution in Keap1 mRNA expression level in pancreas, kidney, and liver. From the outcome of the current results, it can be inferred that seed extract of A. reticulata exhibits a protective effect in diabetic rats through its anti-diabetic, anti-hyperlipidemic, antioxidant and anti-inflammatory effects and could be considered as a promising treatment therapy in the treatment of diabetes mellitus.</p

Hypoxia-induced mitochondrial fragmentation facilitates cristae remodeling in pancreatic beta cells.

A.Representative transmission electron microscopy (TEM) images of mitochondria in pancreatic beta INS-1E cells treatedas indicated.Scale bars, 0.1 μm.B-D. Mitochondrial length (B), cristae width (C) and cristae density(number/mitochondrial length)(D)were quantitatively analyzedin pancreatic beta INS-1E cells treatedas indicated. Over twenty mitochondria were randomly selected for quantitative analysis in each treatment group.All experiments were repeated three times.*P< 0.05.</p

Mdivi-1 exhibits a therapeutic effect on hypoxia-induced cell death of pancreatic beta cells.

A.Mitochondrial morphologyanalysis by confocal microscopy in pancreatic beta INS-1E cells treated with Mdivi-1 (50 μM) or DMSO for 12 h as indicated.Scale bars, 10 μm.B.Cell viability analysis by MTS assay in pancreatic beta INS-1E cells treated with Mdivi-1or DMSO as indicated. C.Detection of apoptosis by flow cytometry in pancreatic beta INS-1E cellstreated with Mdivi-1 or DMSO as indicated. D. TUNEL staining in pancreatic beta INS-1E cells treated with Mdivi-1 or DMSO as indicated.Scale bars, 50 μm.All experiments were repeated three times.*P< 0.05.</p

HIF-1α activation and subsequently DRP1(S616) phosphorylationis involved in hypoxia-induced mitochondrial fragmentation in pancreatic beta cells.

<p><b>A-B.</b> Expression levels of MFN1, MFN2, OPA1, DRP1, Fis1, and MFF were detected by RT–PCR (A) and western blot (B)analysis in pancreatic beta INS-1E cellswith treatment as indicated. <b>(C)</b>Top: Levels of HIF-1α and phosphorylated DRP1 (S637)or(S616)were detected by western blot analysis in pancreatic beta INS-1E cellswith treatment as indicated.Bottom: Western blot scanning densitometry for three independent experiments.Blots were probed for β-actin to ensure equal proteinloading. *P<0.05.<b>(D)</b>Top: Levels of DRP1 in total lysates and mitochondrial fraction were examined by western blot analysis in pancreatic beta INS-1E cellswith treatment as indicated. VDACserves as loading controls. Mito: mitochondria.Bottom: Western blot scanning densitometry for three independent experiments.Blots were probed for β-actinor VDAC respectively to ensure equal proteinloading. *P<0.05.All experiments were repeated three times.</p

Hypoxia induces mitochondrial fragmentation in pancreatic beta INS-1E cells.

<p><b>A.</b> Representative confocal microscope images of the mitochondrial in pancreatic beta INS-1E cellsafter treatment with hypoxia at different oxygen levels (3% or 1% O₂ for 24 h) as indicated. Scale bars, 10 μm.<b>B.</b>The proportion of pancreatic beta INS-1E cellswith tubulated, intermediate, and fragmented mitochondria was quantified. <b>C.</b> Representative transmission electron microscopy (TEM) images of the mitochondrialin INS-1Ecells treated as indicated. Scale bars, 0.5 μm. <b>D.</b>Image J was used for mitochondrial length quantification in INS-1Ecells treated as indicated.All experiments were repeated three times.*<i>P</i>< 0.05.</p

Mitochondrial fragmentation is involved inpancreatic beta cell death in hypoxia conditions.

<p><b>A.</b>Detection of DRP1 (S616) and mitochondrial-fractional DRP1by western blot analysis in pancreatic beta INS-1E cellswith treatment as indicated. <b>B.</b>Detection of apoptosis by flow cytometry in pancreatic beta INS-1E cellswith treatment as indicated. <b>C.</b>Western blot analysis for levels of cytochrome c inpancreatic beta INS-1E cellswith treatment as indicated. β-actin and VDAC were used as loading controls for cytoplasm and mitochondria, respectively. Cyto, cytoplasm; Mito, mitochondrial. <b>D.</b>Western blot analysis for levels ofcleaved Caspase-9 and Caspase-3 inpancreatic beta INS-1E cellswith treatment as indicated. <b>E.</b>TUNEL staining in pancreatic beta INS-1E cells from type 2 diabetes animal models. Blue: Hoechst 33342; Green: TUNEL positive nucleus. All experiments were repeated three times.*<i>P</i>< 0.05.</p