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. 2018 Feb;21(2):165-174.
doi: 10.22038/IJBMS.2017.24801.6167.

Evaluation of TNF-α and IL-6 mRNAs expressions in visceral and subcutaneous adipose tissues of polycystic ovarian rats and effects of resveratrol

Mahnaz Ghowsi  1 , Homayoun Khazali  1 , Sajjad Sisakhtnezhad  2
Affiliations

Evaluation of TNF-α and IL-6 mRNAs expressions in visceral and subcutaneous adipose tissues of polycystic ovarian rats and effects of resveratrol

Mahnaz Ghowsi et al. Iran J Basic Med Sci. 2018 Feb.

Abstract

Objectives: Some studies suggest that chronic low-grade inflammation is involved in insulin resistance in polycystic ovary syndrome (PCOS). This study assessed possible involvement of alteration in expression of two pro-inflammatory factors, tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in adipose tissues of PCOS rats in the impairment of insulin actions. Also, effects of resveratrol as an anti-inflammatory agent were investigated.

Materials and methods: Fifteen female Wistar rats (21 days old) were divided into three groups (n=5): I) Control, II) PCO-model-saline: served as PCOS rats and to induce PCOS, received subcutaneously testosterone enanthate 1 mg/100 g body weight subcutaneously for 35 days, III) PCO-model-resveratrol, after receiving testosterone, received resveratrol 10 mg/kg intraperitoneally for 28 days. The expression of Tnf-α and Il-6 mRNAs in adipose tissues was determined by the qRT-PCR method.

Results: The Il-6 mRNA expression in the visceral adipose tissue of PCOS rats was increased in comparison to controls (P<0.05). Tnf-α and Il-6 mRNA expression in visceral and subcutaneous adipose tissues of polycystic ovarian rats was similar to controls. The expression of Tnf-α mRNA in subcutaneous adipose tissue and Tnf-α and Il-6 mRNAs in the visceral adipose tissue of the PCO-model-resveratrol group were lower than PCOS rats (P<0.05).

Conclusion: Increased expression of Il-6 mRNA in the visceral adipose tissue of polycystic ovarian rats may be one cause of insulin resistance observed in them and resveratrol as an anti-inflammatory and anti-hyperglycemic agent may decrease the risk of diabetes by reduction of expression of pro-inflammatory cytokines TNF-α and IL-6 in PCOS patients.

Keywords: Adipose tissue IL-6; Polycystic ovary – syndrome; Rat; Resveratrol; TNF-α.

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Figures

Figure 1
Figure 1
The flow chart of the experimental protocol. The abbreviations represent the following: Control: Normal control rats; PCO-model-saline: these rats received testosterone enanthate 1 mg/100 g body weight subcutaneously for 35 days once daily. Then, saline was injected into them intraperitoneally for 28 days once daily; PCO-model-resveratrol: these rats received testosterone enanthate 1 mg/100 g body weight subcutaneously for 35 days once daily. Then, resveratrol 10 mg/kg was injected into them intraperitoneally for 28 days once daily
Figure 2
Figure 2
The ovarian sections of rats in the PCO-model-saline groups (A) and in the control group (B) (magnification ×40). The abbreviations represent the following: control: Normal control rats; PCO-model-saline: these rats served as models of PCOS and received testosterone enanthate 1 mg/100 g body weight subcutaneously for 35 days once daily. Then, saline was injected into them intraperitoneally for 28 days once daily. The ovaries in the control group had normal morphology with corpora lutea and antral follicles. The ovaries in PCO-model-saline group displayed cystic follicles and atretic follicles with no corpora lutea. CL: Corpus luteum; CF: Cystic follicle; AF: Antral follicle
Figure 3
Figure 3
The relative expression levels of Tnf-α (A) and Il-6 (B) mRNAs in the visceral adipose tissue of three groups. The expression values were normalized to β-actin and were compared using the 2^-ΔCT method. The data are represented as means for 5 animals per group with the standard errors of the means. Statistics were by one-way ANOVA followed by the Tukey’s post hoc test. P-values less than 0.05 were considered statistically significant. ΨΨ: P<0.01 vs. the PCO-model-saline group. ***: P<0.001 vs. the control group; ΨΨΨ: P<0.001 vs. the PCO-model-saline group
Figure 4
Figure 4
The relative expression levels of Tnf-α (A) and Il-6 (B) mRNAs in subcutaneous adipose tissues of three groups. The expression values were normalized to β-actin and were compared using the 2^-ΔCT method. The data are represented as means for 5 animals per group with the standard errors of the means. Statistics were by one-way ANOVA followed by the Tukey’s Post hoc test. P_values less than 0.05 were considered statistically significant. Ψ: P<0.05 vs. the PCO-model-saline group
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