Chitosan-Stabilized Selenium Nanoparticles and Metformin Synergistically Rescue Testicular Oxidative Damage and Steroidogenesis-Related Genes Dysregulation in High-Fat Diet/Streptozotocin-Induced Diabetic Rats

Abstract

Background: this study examined the metformin (MF) and/or chitosan stabilized selenium nanoparticles (CH-SeNPs) efficacy to alleviate the male reproductive function impairment in a high-fat diet feed with low-dose streptozotocin (HFD/STZ) induced type 2 diabetes mellitus (T2DM) diabetic rat model.

Methods: control non-diabetic, HFD/STZ diabetic, HFD/STZ+MF, HFD/STZ+CH-SeNPs, and HFD/STZ+MF+CH-SeNPs rat groups were used. After 60 days, semen evaluation, hormonal assay, enzymatic antioxidant, lipid peroxidation, testis histopathology, and the steroidogenesis-related genes mRNA expressions were assessed.

Results: in the HFD/STZ diabetic rats, sperm count and motility, male sexual hormones, and testicular antioxidant enzymes were significantly reduced. However, sperm abnormalities and testicular malondialdehyde were significantly incremented. The steroidogenesis-related genes, including steroidogenic acute regulatory protein (StAr), cytochrome11A1 (CYP11A1), cytochrome17A1 (CYP17A1), and hydroxysteroid 17-beta dehydrogenase 3 (HSD17B3), and the mitochondrial biogenesis related genes, including peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGCα) and sirtuin (SIRT), were significantly downregulated in the HFD/STZ diabetic rats. However, CYP19A1mRNA expression was significantly upregulated. In contrast, MF and/or CH-SeNPs oral dosing significantly rescued the T2DM-induced sperm abnormalities, reduced sperm motility, diminished sexual hormones level, testicular oxidative damage, and steroidogenesis-related genes dysregulation. In the MF and CH-SeNP co-treated group, many of the estimated parameters differ considerably from single MF or CH-SeNPs treated groups.

Conclusions: the MF and CH-SeNPs combined treatment could efficiently limit the diabetic complications largely than monotherapeutic approach and they could be considered a hopeful treatment option in the T2DM.

Keywords: chitosan-stabilized selenium nanoparticles; male fertility; metformin; oxidative stress; steroidogenesis related genes; testicular dysfunction; type 2 diabetes mellitus.

Figure 1

Effect of metformin (MF) and/or chitosan stabilized nanoparticles (CH-SeNPs) oral dosing for 60 days on sperm characteristics including (A) sperm motility, (B) live sperms, (C) sperm concentration, and (D) sperm abnormalities in high-fat diet/streptozotocin (HFD/STZ) diabetic male rats. Data expressed as mean ± SE, n = 15 for each group. Each bar carrying different letters (a–d) was significantly different at p < 0.05.

Figure 2

Effect of metformin (MF) and/or chitosan stabilized nanoparticles (CH-SeNPs) oral dosing for 60 days on sexual hormonal variables including (A) testosterone (TES), (B) follicle-stimulating hormone (FSH) (C), luteinizing hormone (LH), and (D) estradiol (E2) levels in the serum of HFD/STZ diabetic male rats. Data are expressed as mean ± SE, n = 15 for each group. Each bar carrying different letters (a–e) was significantly different at p < 0.05.

Figure 3

Effect of metformin (MF) and/or chitosan stabilized nanoparticles (CH-SeNPs) oral dosing for 60 days on (A) superoxide dismutase (SOD) (B), catalase (CAT), and (C) malondialdehyde (MDA) levels in the testicular tissues of HFD/STZ diabetic male rats. Data are expressed as mean ± SE, n = 15 for each group. Each bar carrying different letters (a–e) was significantly different at p < 0.05.

Figure 4

Effect of metformin (MF) and/or chitosan stabilized nanoparticles (CH-SeNPs) oral dosing for 60 days on mRNA expression of (A) steroidogenic acute regulatory protein (StAr), (B) cytochrome11A1 (CYP11A1), (C) cytochrome17A1 (CYP17A1) and (D) hydroxysteroid 17-beta dehydrogenase 3 (HSD17B3) in the testicular tissues of HFD/STZ diabetic male rats. Data are expressed as mean ± SE, n = 15 for each group. Each bar carrying different letters (a–d) was significantly different at p < 0.05.

Figure 5

Effect of metformin (MF) and/or chitosan stabilized nanoparticles (CH-SeNPs) oral dosing for 60 days on mRNA expression of (A) aromatase gene (CYP19A1), (B) peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α), and (C) Silent information regulator type-1 (SIRT1) in the testicular tissues of HFD/STZ diabetic male rats. Data expressed as mean ± SE, n = 15 for each group. Each bar carrying different letters (a–d) was significantly different at p < 0.05.

Figure 6

Representative photomicrograph of H&E stained testicular tissue sections showing normal histological picture in the control rats (A,B). HFT/STZ testes showing congestion (red arrowhead), vacuolated germinal epithelium (blue arrowhead), necrotic germinal epithelium, (yellow arrow), interstitial fibrosis (black arrowhead), and redundant basal lamina (arrow) (C,D). HFT/STZ+MF testes showing germ cell depletion (black ellipse), spermatid retention (yellow ellipse), thickened basal lamina (yellow arrowhead), interstitial edema (blue arrowhead), fibrosis (black arrowhead), and congestion (red arrowhead) (E,F). HFT/STZ+CH-SeNPs testes showing congestion (red arrowhead), multinucleated giant cell s (black arrowheads) and redundant basal lamina (arrow) (G,H). HFT/STZ+MF+CH-SeNPs testes showing congestions (red arrowheads), desquamated germinal epithelium (black arrowhead), and sperm stasis (ellipse) (I,J). Scale bar is 100 microns for (A,C,E,G,I), and 20 microns for (B,D,F,H,J).