Ann Lab Med.  2020 Sep;40(5):390-397. 10.3343/alm.2020.40.5.390.

Cytokines, Angiogenesis, and Extracellular Matrix Degradation are Augmented by Oxidative Stress in Endometriosis

• Nanda A
• K. T
• Banerjee P
• Dutta M
• Wangdi T
• Sharma P
• Chaudhury K
• Jana SK

• KMID: 2502569
• DOI: http://doi.org/10.3343/alm.2020.40.5.390

Abstract

Background
The effect of the interplay among inflammation, angiogenesis, extracellular matrix (ECM) degradation, and oxidative stress (OS) on the pathogenesis of endometriosis remains unclear. Previously, we demonstrated the role of OS in endometriosis. Here, we performed a comprehensive investigation of several molecules involved in inflammation, angiogenesis, and ECM degradation in women with endometriosis to study their interplay with OS.
Methods
Blood samples were collected from women with endometriosis (N=80), as well as from women with tubal factor infertility as controls (N=80). Interleukin (IL)-1β, tumor necrosis factor-alpha, interferon-gamma, transforming growth factor-beta, IL-4, -10, -2, -6, -8, vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP)-2, -9, tissue inhibitor of metalloproteinases (TIMP)-1, -2, and cyclooxygenase (COX)-2 levels in serum samples were measured using an ELISA. Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) in peripheral blood mononuclear cells was measured using flow cytometry.
Results
Cytokines, VEGF, MMPs, and COX-2 were significantly higher and TIMPs were significantly lower in patients with endometriosis. Multivariate statistical analysis indicated that IL-10 was the most significant variable capable of discriminating endometriosis samples from controls.
Conclusions
Deregulation of NF-κB activation by OS affects the expression of various cytokines in endometriosis. Elevated cytokine levels further up-regulate IL-10, which subsequently activates the MMPs, leading to excessive ECM degradation and angiogenesis. Moreover, IL-10 emerged as the most important molecule involved in the pathogenesis of endometriosis. Measurement of these molecules may help in better management of the patients with endometriosis.

Keyword

Oxidative stress (OS); Endometriosis; Extracellular matrix; Cytokines

Figure

• 

  Fig. 1 Flow cytometry analysis of NF-κB activation. (A) Percentage of cells with NF-κB (p50 and p65) activation as measured by flow cytometry. (B) Graphical representation of cells with NF-κB activation as measured by flow cytometry. Abbreviations: Cntrl, control; Endo, endometriosis.

• 

  Fig. 2 Scores scatter plot of (A) PCA showing discrimination between endometriosis and controls, (B) PLS-DA showing improved discrimination between the two groups. (C) PLS-DA model cross-validation by permutation test statistics showing that the R2 (0.986) and Q2 (0.976) values of the original model are significantly greater than those of the 100 permutated models, indicating a good predictive ability. Variable extraction using (D) loading and (E) VIP plots showing that IL-10 is the most important variable for class separation. Abbreviations: PCA, principal-component analysis; PLS-DA, partial least square discriminate analysis; VIP, variable importance plot; IL, interleukin; TNF, tumor necrosis factor; IFN, interferon; TGF, transforming growth factor; VEGF, vascular endothelial growth factor; MMP, matrix metalloproteinase; TIMP, tissue inhibitor of metalloproteinase; COX, cyclooxygenase.

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