Yang Ming-Gen, Xu Zhen-Qiang
Department of Urology, Zhangzhou Hospital Affiliated to Fujian Medical University, Zhangzhou, Fujian 363000, China.
Zhonghua Nan Ke Xue. 2021 Oct 20;27(10):867-875.
To investigate the impact of macrophage-induced immune inflammation on the proliferation and apoptosis of BPH cells and its possible mechanism.
Macrophages were stimulated with phorbol myristate acetate, co-cultured with BPH-1 cells, and then treated with the androgen receptor (AR) inhibitor or anti-CD40L antibody. The immunohistochemical biomarkers of the T lymphocytes (CD4 and CD8), B lymphocyte (CD20) and macrophages (CD68), AR, CD40/CD40L, and inflammatory factors IL-1, IL-6 and TNF-α were measured before and after treatment. The proliferation and apoptosis of the cells were observed by MTT assay, colony-forming assay and flow cytometry, and the expressions of cell apoptosis- and MAPK signaling pathway-related proteins were determined by qRT-PCR and Western blot.
Significantly increased proliferation and decreased apoptosis of the cells, up-regulated expressions of Bcl-2, IL-1, IL-6, TNF-α, AR, CD40 and CD40L, and down-regulated expression of Bax were observed in the BPH-1 cells co-cultured with macrophages (the M-BPH-1 group) compared with those in the blank control (B-BPH-1) group (P < 0.01). In comparison with the BPH-1 cells treated with normal saline, those treated with either low-dose CD40L (L-CD40L) or high-dose CD40L (H-CD40L) showed markedly inhibited proliferation, increased apoptosis, up-regulated expression of Bax, and down-regulated expressions of Bcl-2, IL-1, IL-6 and TNF-α (P < 0.01), and those in the low- and high-dose AR (L-AR and H-AR) inhibitor groups exhibited remarkably reduced proliferation, increased apoptosis, down-regulated expressions of Bcl-2, IL-1, IL-6 and TNF-α, and up-regulated expression of Bax (P < 0.01). The phosphorylation levels of JNK, ERK and P38 were significantly elevated in the M-BPH-1 group, but declined in the H-CD40L and the H-AR inhibitor groups compared with those in the B-BPH-1 group, all in a concentration-dependent manner (P < 0.01).
Macrophage-induced immune inflammation regulates AR and CD40/CD40L expressions and promotes the proliferation and inhibits the apoptosis of BPH-1 cells by activating the MAPK signaling pathway. /.
探讨巨噬细胞诱导的免疫炎症对前列腺增生（BPH）细胞增殖和凋亡的影响及其可能机制。
用佛波酯刺激巨噬细胞，与BPH-1细胞共培养，然后用雄激素受体（AR）抑制剂或抗CD40L抗体处理。在处理前后检测T淋巴细胞（CD4和CD8）、B淋巴细胞（CD20）和巨噬细胞（CD68）、AR、CD40/CD40L以及炎性因子IL-1、IL-6和TNF-α的免疫组化生物标志物。通过MTT法、集落形成试验和流式细胞术观察细胞的增殖和凋亡，并用qRT-PCR和蛋白质免疫印迹法检测细胞凋亡及丝裂原活化蛋白激酶（MAPK）信号通路相关蛋白的表达。
与空白对照组（B-BPH-1组）相比，与巨噬细胞共培养的BPH-1细胞（M-BPH-1组）细胞增殖显著增加，凋亡减少，Bcl-2、IL-1、IL-6、TNF-α、AR、CD40和CD40L表达上调，Bax表达下调（P<0.01）。与用生理盐水处理的BPH-1细胞相比，用低剂量CD40L（L-CD40L）或高剂量CD40L（H-CD40L）处理的细胞增殖明显受抑制，凋亡增加，Bax表达上调，Bcl-2、IL-1、IL-6和TNF-α表达下调（P<0.01）；低剂量和高剂量AR（L-AR和H-AR）抑制剂组细胞增殖显著降低，凋亡增加，Bcl-2、IL-1、IL-6和TNF-α表达下调，Bax表达上调（P<0.01）。与B-BPH-1组相比，M-BPH-1组JNK、ERK和P38的磷酸化水平显著升高，而H-CD40L组和H-AR抑制剂组则下降，均呈浓度依赖性（P<0.01）。
巨噬细胞诱导的免疫炎症通过激活MAPK信号通路调节AR和CD40/CD40L的表达，促进BPH-1细胞的增殖并抑制其凋亡。
