Changes in MCP-1, HGF, and IGF-1 expression in endometrial stromal cells, PBMCs, and PFMCs of endometriotic women following 1,25(OH)2D3 treatment

Abstract

1,25(OH)2D3 has anti-inflammatory and growth inhibitory effects. Our study explored the effect of 1,25(OH)2D3 treatment on the expression of monocyte chemotactic protein-1 (MCP-1), hepatocyte growth factor (HGF), and insulin-like growth factor-1 (IGF-1) by peripheral blood mononuclear cells (PBMCs), peritoneal fluid mononuclear cells (PFMCs), endometrial stromal cells (ESCs), and its effect on the proliferation of PBMCs and PFMCs of patients with endometriosis compared with controls. PBMCs, PFMCs, and ESCs were obtained from 10 endometriosis patients and 10 non-endometriotic individuals. After treating cells with 0.1 μM of 1,25(OH)2D3 for 6, 24, and 48 h, the gene and protein expression of mentioned factors were evaluated by real-time PCR and ELISA methods, respectively. 1,25(OH)2D3 treatment significantly reduced the protein expression of MCP-1, HGF, and IGF-1 in PBMCs and PFMCs of endometriotic patients at 48 h (p < 0.05-<0.01). Also, this treatment significantly reduced MCP-1, HGF, and IGF-1 gene and/or protein expression in EESCs and EuESCs at 24 and 48 h (p < 0.05-<0.01). 1,25(OH)2D3 treatment also reduced the proliferation of PBMCs and PFMCs of endometriotic patients compared with controls (p < 0.01). 1,25(OH)2D3 can be considered as a potentially effective agent in the prevention and treatment of endometriosis along with other therapies.

Keywords: 1,25(OH)2D3; HGF; IGF-1; MCP-1; endometrial stromal cells; endometriosis; mononuclear cells.

FIGURE 1

The gene and protein expression of MCP‐1 by PBMCs, PFMCs, and ESCs after treatment with 1,25(OH)2D3. PBMCs of endometriosis patients and control participants (n = 10), PFMCs of endometriosis patients and control participants (n = 8), EESCs (n = 8), EuESCs (n = 10), and CESCs (n = 10) were treated with 0.1 μM 1,25(OH)2D3 at 6, 24, and 48 h. Results were analysed using a non‐parametric test. (A) Treatment of PBMCs, (B) Treatment of PFMCs, and (C) Treatment of ESCs. (a) The gene expression of MCP‐1 at 6 h (D₃ +/−), (b) The gene expression of MCP‐1 at 24 h (D₃ +/−), (c) The gene expression of MCP‐1 at 48 h (D₃ +/−), (d) protein production of MCP‐1 at 6 h (D₃ +/−), (e) protein production of MCP‐1 at 24 h (D₃ +/−), and (f) protein production of MCP‐1 at 48 h (D₃ +/−). Data were represented as mean ± SEM and min to max. *p < 0.05 and **p < 0.01. CESCs, control endometrial stromal cells; EESCs, ectopic endometrial stromal cells; ESCs, endometrial stromal cells; EuESCs, eutopic endometrial stromal cells; MCP‐1, monocyte chemoattractant protein‐1; PBMCs, peripheral blood mononuclear; PFMCs, peritoneal fluid mononuclear cells

FIGURE 2

The gene and protein expression of HGF by PBMCs, PFMCs, and ESCs after treatment with 1,25(OH)2D3. PBMCs of endometriosis patients and control participants (n = 10), PFMCs of endometriosis patients and control participants (n = 8), EESCs (n = 8), EuESCs (n = 10), and CESCs (n = 10) were treated with 0.1 μM 1,25(OH)2D3 at 6, 24, and 48 h. Results were analysed using a non‐parametric test. (A) Treatment of PBMCs, (B) Treatment of PFMCs, and (C) Treatment of ESCs. (a) The gene expression of HGF at 6 h (D₃ +/−), (b) The gene expression of HGF at 24 h (D₃ +/−), (c) The gene expression of HGF at 48 h (D₃ +/−), (d) protein production of HGF at 6 h (D₃ +/−), (e) The protein production of HGF at 24 h (D₃ +/−), and (f) The protein production of HGF at 48 h (D₃ +/−). Data were represented as mean ± SEM and min to max. *p < 0.05 and **p < 0.01. CESCs, control endometrial stromal cells; EESCs, ectopic endometrial stromal cells; ESCs, endometrial stromal cells; EuESCs, eutopic endometrial stromal cells; HGF, Hepatocyte growth factor; PBMCs, peripheral blood mononuclear; PFMCs, peritoneal fluid mononuclear cells

FIGURE 3

The gene and protein expression of IGF‐1 by PBMCs, PFMCs, and ESCs after treatment with 1,25(OH)2D3. PBMCs of endometriosis patients and control participants (n = 10), PFMCs of endometriosis patients and control participants (n = 8), EESCs (n = 8), EuESCs (n = 10), and CESCs (n = 10) were treated with 0.1 μM 1,25(OH)2D3 at 6, 24, and 48 h. Results were analysed using a non‐parametric test. (A) Treatment of PBMCs, (B) Treatment of PFMCs, and (C) Treatment of ESCs. (a) The gene expression of IGF‐1 at 6 h (D₃ +/−), (b) The gene expression of IGF‐1 at 24 h (D₃ +/−), (c) The gene expression of IGF‐1 at 48 h (D₃ +/−), (d) protein production of IGF‐1 at 6 h (D₃ +/−), (e) protein production of IGF‐1 at 24 h (D₃ +/−), and (f) protein production of IGF‐1 at 48 h (D₃ +/−). Data were represented as mean ± SEM and min to max. *p < 0.05 and **p < 0.01. CESCs, control endometrial stromal cells; EESCs, ectopic endometrial stromal cells; ESCs, endometrial stromal cells; EuESCs, eutopic endometrial stromal cells; IGF‐1, Insulin growth factor‐1; PBMCs, peripheral blood mononuclear; PFMCs, peritoneal fluid mononuclear cells

FIGURE 4

(A) The proliferation rate of PBMCs and (B) PFMCs of endometriosis and control participants after 1,25(OH)2D3 treatment. Results reported as mean ± SD. PBMCs of endometriosis patients and control participants (n = 10), PFMCs of endometriosis patients and control participants (n = 8). **p < 0.01. PBMCs, peripheral blood mononuclear cells; PFMCs, peritoneal fluid mononuclear cells