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. 2022;13(4):489-493.
doi: 10.30466/vrf.2022.544355.3314. Epub 2022 Dec 15.

Evaluation of histopathological changes and exosomal biogenesis in pulmonary tissue of diabetic rats

Aref Delkhosh  1 , Rahim Hobbenaghi  1 , Reza Rahbarghazi  2   3 , Mahdi Ahmadi  2 , Jafar Rezaie  4
Affiliations

Evaluation of histopathological changes and exosomal biogenesis in pulmonary tissue of diabetic rats

Aref Delkhosh et al. Vet Res Forum. 2022.

Abstract

Diabetes mellitus is one of the leading causes of death globally. The development of cellular injuries and impaired energy metabolism are involved in the pathogenesis of diabetes mellitus, leading to severe diabetic complications in different tissues such as the pulmonary tissue. Autophagy is a double-edged sword mechanism required for maintaining cell survival and homeostasis. Any abnormalities in autophagic response can lead to the progression of several diseases. Here, we aimed to assess the effect of diabetic conditions on the autophagic response and exosome secretion in a rat model of type 2 diabetes mellitus. The experimental diabetic group received 45.00 mg kg-1 streptozocin (STZ) dissolved in 0.10 M sodium citrate. After 4 weeks, we monitored autophagic response and exosome biogenesis in the pulmonary tract using immunohistochemistry (IHC) and Real-time polymerase chain reaction analyses, respectively. Histological examination revealed the interstitial bronchopneumonia indicating enhanced immune cell infiltration into the pulmonary parenchyma. Immunohistochemistry staining displayed an enhanced autophagic response through the induction of microtuble-associated protein light chain 3 (LC3) and protein sequestosome 1 (P62) compared to the control rats. These changes coincided with significant induction of tetraspanin CD63 in STZ-induced diabetic rats relative to control rats. In conclusion, a diabetic condition can increase the autophagic response in pulmonary tissue. The accumulation of P62 in the pulmonary niche exhibits an incomplete autophagic response. The abnormal autophagy response can increase pulmonary cell sensitivity against injuries.

Keywords: Animal model; Autophagy; Diabetes mellitus; Exosome; Histology.

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Conflict of interest statement

Authors declare there is no conflict of interest related to this study.

Figures

Fig. 1
Fig. 1
A) Control group; B) Bright-field staining revealed enhanced immune cell infiltration into the pulmonary niche after the induction of inflammatory cells. These features led to the thickening of alveolar septa (black arrows) and generation of several emphysematous foci (asterisk), (H & E staining; bars = 250 µm). C) Control group, and D) showed type I collagen deposition (blue-colored areas; yellow arrows) within the pulmonary parenchyma in the diabetic group (Masson’s trichrome staining; bars = 50 and 20 µm, respectively)
Fig. 2
Fig. 2
Immunohistochemical staining of in pulmonary tissue. Black arrows indicate LC3 autophagic marker in A) Control group;  B) Diabetic group. Data showed promotion of LC3 factors after the induction of diabetic conditions (bars = 100 µm). Intra-cellular accumulation of P62 (Red arrows) showed an incomplete autophagy response in C) Control group, and D) Diabetic group (bars = 100 µm)
Fig. 3
Fig. 3
Real-time polymerase chain reaction analysis of CD63 in pulmonary tissue after diabetes mellitus induction. * indicates statistically significant up-regulation of CD63 after diabetes mellitus induction (p < 0.01)
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