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. 2021 Jun 7;11(1):11924.
doi: 10.1038/s41598-021-91459-x.

Effect of antioxidant treatment with n-acetylcysteine and swimming on lipid expression of sebaceous glands in diabetic mice

Geovane Ribeiro Santos  1   2   3 , Marcelo Rodrigues Cunha  1 , Eduardo José Caldeira  1 , Ewerton Alexandre Galdeano  1 , Raphael Cruz Seabra Prudente  4 , Clóvis Antonio Lopes Pinto  5   6   7
Affiliations

Effect of antioxidant treatment with n-acetylcysteine and swimming on lipid expression of sebaceous glands in diabetic mice

Geovane Ribeiro Santos et al. Sci Rep. .

Abstract

The sebaceous gland (SG) is involved in different inflammatory, infectious and neoplastic processes of the skin and can be related to specific diseases, e.g., diabetes mellitus. Sometimes, the histological diagnosis requires complementary tests due to the ability of diseases to mimic other tumors. We evaluated the sebaceous gland density in Non-obese diabetic mice to analyze the N-acetylcystein effects and swimming exercise treatment in sebaceous glands healing, using specific staining in histochemistry and immunohistochemistry reactions in the identification of the lipid expression in the sebaceous gland. We investigated the intracytoplasmic lipid expression and analysis of gland density from SG in dorsal skin samples from the Non-obese diabetic (NOD mice) and diabetic animals submitted to antioxidant treatment and physical exercise. For histological analysis of the sebaceous glands, specific staining in histochemistry with sudan black and immunohistochemistry reaction with adipophilin were used in the evaluation. Statistical analysis showed significant proximity between the values of the control group and the diabetic group submitted to the swimming exercise (DS group) and similar values between the untreated diabetic group (UD group) and diabetic group treated with the antioxidant N-acetylcysteine (DNa group), which did not prevent possible differences where p < 0.01. Adipophilin (ADPH) immunohistochemistry permitted more intense lipid staining in SGs, the preservation of the SG in the control group, and a morphological deformed appearance in the UD and DNa groups. However, weak morphological recovery of the SG was observed in the DS-Na group, being more expressive in the DS group. In conclusion, the groups submitted to physical exercises showed better results in the recovery of the analyzed tissue, even being in the physiological conditions caused by spontaneous diabetes.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
In (A–D) photomicrographs of mice tissues sections, evaluated in different enlargement in the microscopy, allowed us to observe how much the immunohistochemical technique by ADPH becomes essential, due to its sensitivity, to allow sebaceous differentiation in paraffin-embedded material, as demonstrated in the scheme represented in (E). In (F–H), we have photomicrographs of the sections of the mice’s frozen tissue stained with Sudan Black. Note the presence of black spots, provided by the labeling of the many lipids present in fresh tissue. This technique, when evaluated in a different objective, does not allow the sebaceous differentiation, as demonstrated in the scheme in (I).
Figure 2
Figure 2
Photomicrographs (×600) of paraffinized tissue sections of mice analyzed by ADPH immunohistochemistry. (AI) Notice the fine staining of fat and intracytoplasmic lipid expression in sebaceous cells (yellow arrow). This technique allows us to identify cellular limits (red arrow) and the presence of nuclei (green arrow), as well as expressive differences in the sebaceous glands’ size. (AI) Control group. (BI) UD group. (CI) DS group. (DI) DNa group. (EI) DS-Na group. (AII,BII,CII,DII,EII): sebaceous gland area (µm2) box plot evaluated in control group, UD group, DS group, DNa group and DS-Na group, respectively. (AIII,BIII,CIII,DIII,EIII) Average of the sebaceous gland area (µm2) evaluated in the control group, UD group, DS group, DNa group and DS-Na group, respectively.
Figure 3
Figure 3
Photomicrographs (×200) of frozen mice tissue sections stained with Sudan Black. Note the presence of a fat spot and intracytoplasmic lipid expression in sebaceous cells. The technique permitted visualization of differences in the size of the sebaceous glands (black circle). (A) Control group. (B) UD group. (C) DS group. (D) DNa group. (E) DS-Na group.
Figure 4
Figure 4
Photomicrographs (×400) of paraffinized tissue sections of control mice allowing, in (A), the cellular limits identification (red arrow) and the presence of nuclei (green arrow) by HE staining. In (B), notice the good expression of intracytoplasmic lipids in sebaceous cells (yellow arrow), allowing the identification of cellular limits (red arrow) and the presence of nuclei (green arrow) by ADPH immunohistochemistry. (C) Shows frozen tissue sections of the control group mice stained with Sudan Black. Notice the presence of tissue fat spots and lipid expression in sebaceous glands (black circle).
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References

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