Department of Urology, Hebei Province of Chinese Medicine/Affiliated Hospital of Hebei University of Chinese Medicine, No. 389, Zhongshan East Road, Shijiazhuang, 050000, Hebei Province, People's Republic of China.
Inflamm Res. 2022 Apr;71(4):449-460. doi: 10.1007/s00011-022-01544-8. Epub 2022 Mar 13.
Chronic nonbacterial prostatitis (CNP) has remained one of the most prevalent urological diseases, particularly in older men. Dihydroartemisinin (DHA) has been identified as a semi-synthetic derivative of artemisinin that exhibits broad protective effects. However, the role of DHA in inhibiting CNP inflammation and prostatic epithelial cell proliferation remains largely unknown.
CNP animal model was induced by carrageenan in C57BL/6 mouse. Enzyme linked immunosorbent assay (ELISA), Real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot were used to examine inflammatory cytokines and proliferation genes expression. Immunofluorescence and immunochemistry staining were used to detect and E2F7 expression. Human prostatic epithelial cells (HPECs) and RWPE-1 was induced by lipopolysaccharide (LPS) to mimic CNP model in vitro. Cell proliferation was determined using MTS assay.
DHA significantly alleviated the rough epithelium and inhibited multilamellar cell formation in the prostatic gland cavity and prostatic index induced by carrageenan. In addition, DHA decreased the expression of TNF-α and IL-6 inflammatory factors in prostatitis tissues and in LPS-induced epithelial cells. Upregulation of transcription factor E2F7, which expression was inhibited by DHA, was found in CNP tissues, human BPH tissues and LPS-induced epithelial cells inflammatory response. Mechanically, we found that depletion of E2F7 by shRNA inhibited epithelial cell proliferation and LPS-induced inflammation while DHA further enhance these effects. Furthermore, HIF1α was transcriptional regulated by E2F7 and involved in E2F7-inhibited CNP and cellular inflammatory response. Interestingly, we found that inhibition of HIF1α blocks E2F7-induced cell inflammatory response but does not obstruct E2F7-promoted cell growth.
The results revealed that DHA inhibits the CNP and inflammation by blocking the E2F7/HIF1α pathway. Our findings provide new evidence for the mechanism of DHA and its key role in CNP, which may provide an alternative solution for the prevention and treatment of CNP.
慢性非细菌性前列腺炎（CNP）仍然是最常见的泌尿系统疾病之一，尤其是在老年男性中。二氢青蒿素（DHA）已被确定为青蒿素的半合成衍生物，具有广泛的保护作用。然而，DHA 抑制 CNP 炎症和前列腺上皮细胞增殖的作用在很大程度上尚不清楚。
用角叉菜胶诱导 C57BL/6 小鼠建立 CNP 动物模型。酶联免疫吸附试验（ELISA）、实时定量聚合酶链反应（RT-qPCR）和 Western blot 用于检测炎症细胞因子和增殖基因的表达。免疫荧光和免疫组织化学染色用于检测和 E2F7 表达。用人前列腺上皮细胞（HPECs）和 RWPE-1 用脂多糖（LPS）诱导建立体外 CNP 模型。MTS 法测定细胞增殖。
DHA 显著减轻了角叉菜胶诱导的前列腺腺泡粗糙上皮和抑制了多层层状细胞形成和前列腺指数。此外，DHA 降低了前列腺炎组织和 LPS 诱导的上皮细胞中 TNF-α和 IL-6 炎症因子的表达。在 CNP 组织、人 BPH 组织和 LPS 诱导的上皮细胞炎症反应中，发现转录因子 E2F7 的表达上调，而 DHA 抑制了其表达。机制上，我们发现 E2F7 的 shRNA 耗竭抑制了上皮细胞增殖和 LPS 诱导的炎症，而 DHA 进一步增强了这些作用。此外，E2F7 转录调控 HIF1α，并参与 E2F7 抑制的 CNP 和细胞炎症反应。有趣的是，我们发现抑制 HIF1α 可阻断 E2F7 诱导的细胞炎症反应，但不阻碍 E2F7 促进的细胞生长。
结果表明，DHA 通过阻断 E2F7/HIF1α 通路抑制 CNP 和炎症。我们的研究结果为 DHA 的作用机制及其在 CNP 中的关键作用提供了新的证据，这可能为 CNP 的预防和治疗提供一种替代方案。
