Nanoparticles of Costus  speciosus Ameliorate Diabetes-Induced Structural Changes in Rat Prostate through Mediating the Pro-Inflammatory Cytokines IL 6, IL1β and TNF-α

Abstract

Diabetes mellitus is a common global health problem. Among the complications that are frequently associated with DM are the alternation of sexual function and fertility, especially in young men. This study aimed to assess the efficacy of nanoparticles of Costus speciosus (C. speciosus) in preserving the prostatic structure of diabetic rats and to explore the mechanism behind this effect. A model of DM was induced in male albino rats by a single intraperitoneally injection of streptozotocin (STZ, 60 mg/kg body weight). Five groups (n = 10 each) of rats were included in this study: the control, C. speciosus gold nanoparticles-treated (150 mg/kg body weight through gastric intubation for 30 days), untreated diabetic, metformin-treated diabetic (500 mg/kg/day gastric intubation for 30 days) and the C. speciosus-treated diabetic group. The blood glucose, insulin and testosterone levels as well as oxidants/antioxidants status were assessed in the serum. Gene expression of proinflammatory cytokines TNF-α, IL1β and IL-6 were assessed in the prostate homogenate. At the end of the experiment, the rats were sacrificed and the prostate was dissected out and prepared for histopathological and immunohistochemistry study using Ki67 and Bcl-2. C. Speciosus nanoparticles significantly decreased (p = 0.03) the blood glucose level while significantly increasing insulin (p = 0.01) and testosterone (p = 0.04) levels compared to the untreated diabetic rats. Oxidants/antioxidants status was markedly improved after administration of C. speciosus. Prostatic expression of the mRNA of pro-inflammatory cytokines IL-6, IL1β and TNF-α was down-regulated in metformin- and C. speciosus-treated rats. The histological structure of the ventral prostate was preserved in metformin- and C. speciosus-treated diabetic rats with a significantly thicker epithelial cell layer and significant increase immunoexpression in Bcl-2 and Ki67. In conclusion, the protective effect induced by C. speciosus nanoparticles on the prostate of diabetic rats might be directly mediated through the down-regulation of inflammatory cytokines and the up-regulation of antioxidant activity and indirectly mediated through the anti-hyperglycemic effect through enhancing insulin secretion.

Keywords: Bcl-2; Costus speciosus; Ki67; cytokines; diabetes mellitus; inflammation; insulin; nanoparticles; prostate; testosterone.

Figure 1

Graphical scheme of the study design.

Figure 2

(A) Particle size distribution measured by dynamic laser light scattering (DLS) technique. (B) Scanning electron micrograph shows uniform and spherical shape of C. speciosus extract-loaded nanostructured lipid carriers (CSE-NLCs) with no evident signs of aggregation.

Figure 3

Expression of the mRNA of pro-inflammatory cytokines IL-6 (A), IL1β (B) and TNF-α (C) in the ventral prostate of the studied groups was estimated using qRT-PCR. Results are presented as mean ± SD, n = 10. The studied groups were compared using one-way ANOVA test followed by the Bonferroni post-hoc test. Significance is considered at p < 0.05.

Figure 4

Sections of ventral prostates of control and C. speciosus-treated rats show intact prostatic acini with different sizes and shapes and with many epithelial folds while those of untreated diabetic rats show enlarged, widely separated acini with reduced height of acinar epithelium, less epithelial folds and many degenerated epithelial cells (arrow) that appear in the lumen (star) as well as dilated capliraries (interrupted arrow). Ventral prostate of metformin- and C. speciosus-treated diabetic rats show much preserved structure (Haematoxylin and eosin stained sections (A–J). Ki67 and BCL-2 immunoexpression in ventral prostate are shown (K–T). Inserts of the control group show the negative control of the immunostaining.

Figure 5

Prostatic epithelium height (A), immunoexpression of Ki67 (B) and BCL-2 (C) were measured in the ventral prostate of the studied groups. Results are presented as mean ± SD, n = 10. The studied groups are compared using one-way ANOVA test followed by Bonferroni post-hoc test. Significance is considered at p < 0.05.