THP-1 유래 CM 유도 전립선비대 모델에서 Berberis aristata 열수 추출물의 IL-6/STAT3 매개 염증 억제 효과 = Anti-inflammatory Effects of Berberis aristata Hot-Water Extract on IL-6/STAT3 Signaling in a THP-1-Derived CM-Induced Benign Prostatic Hyperplasia Model

다국어 초록 (Multilingual Abstract)

Benign prostatic hyperplasia (BPH) is characterized by epithelial and stromal hyperplasia accompanied by chronic inflammation, in which the IL-6/STAT3 signaling axis plays a central pathogenic role. Natural products with anti-inflammatory potential, such as Berberis aristata extract (BAE), have been suggested as alternative therapeutic candidates; however, their molecular mechanisms in BPH remain unclear. In this study, we aimed to investigate the molecular mechanisms underlying the anti-inflammatory effects of BAE observed in previous in vitro studies.
To elucidate the anti-inflammatory mechanism of BAE, both in vivo testosterone propionate (TP)–induced BPH rat models and in vitro BPH models were employed. STAT3 protein expression in prostate tissues was examined by immunohistochemistry. In vitro, an inflammatory microenvironment in BPH was established by treating normal prostate epithelial (RWPE-1) and stromal cells (WPMY-1) with LPS-stimulated THP-1–derived conditioned medium (CM-LPS). A time-course analysis of IL-6, STAT3, and SOCS3 expression was performed to determine the CM treatment time for subsequent experiments. Based on these findings, co-treatment and pre-treatment experiments were conducted to evaluate the preventive and therapeutic effects of BAE.
In vivo, TP-induced BPH prostates showed strong STAT3 activation and epithelial hyperplasia, whereas BAE reduced STAT3 expression in a dose-dependent manner. Notably, BAE at 200 mg/kg markedly suppressed STAT3 expression and improved tissue architecture, demonstrating efficacy comparable to berberine (BBR) and finasteride (PC). While BBR reduced STAT3 protein, it did not significantly suppress STAT3 mRNA, suggesting a distinct mechanism from BAE.
In vitro, CM-LPS induced a typical IL-6/STAT3 activation pattern, with IL-6 upregulated at 2–4 h and STAT3/SOCS3 elevated at 24 h. BAE modulated this signaling axis in a cell-type- and time-dependent manner. In WPMY-1 cells, BAE prominently suppressed IL-6 expression and downstream STAT3 activation, whereas in RWPE-1 cells, BAE primarily enhanced SOCS3-mediated negative feedback rather than directly reducing IL-6. Under early inflammatory conditions (2 h CM-LPS), BAE significantly reduced IL-6 and increased SOCS3 in both cell types. Pre-treatment experiments confirmed that BAE effectively prevented IL-6/STAT3 activation, particularly in stromal cells.
This study demonstrates that BAE attenuates benign prostatic hyperplasia by modulating the IL-6/STAT3/SOCS3 signaling axis at multiple regulatory levels in both prostatic epithelial and stromal cells. The consistency between in vitro mechanistic findings and in vivo outcomes suggests that BAE may serve as a promising natural agent for mitigating chronic inflammation–driven prostatic hyperplasia.

목차 (Table of Contents)

• Ⅰ. 서론 1
• 연구목적 6
• Ⅱ. 재료 및 방법 7
• 2.1. 실험재료 7
• 2.2. 실험방법 8
• 2.2.1. 실험동물 8
• 2.2.2. 면역조직화학(IHC) 분석 11
• 2.2.3. 조직 RNA 추출 및 Real-time qPCR 11
• 2.2.4. 세포 배양 13
• 2.2.5. 세포 생존율 분석(MTT assay) 13
• 2.2.6. THP-1 대식세포 분화 및 Conditioned medium(CM) 제조 14
• 2.2.7. THP-1 유래 CM 처리에 의한 전립선 세포의 시간별 유전자 발현 변화 14
• 2.2.8. THP-1 대식세포 분화 및 Conditioned medium(CM) 제조 16
• 2.2.9. 세포 RNA 추출 및 Real-time qPCR 21
• 2.2.10. 통계분석 23
• Ⅲ. 결과 및 고찰 24
• 3.1. 실험동물의 전립선 조직에서 BAE의 경구투여의 STAT3 활성 억제 효과 24
• 3.1.1. BAE 투여에 따른 STAT3의 면역조직화학분석 24
• 3.1.2. BAE 투여에 따른 STAT3의 mRNA 발현 변화 26
• 3.2. THP-1 유래 CM 유도 전립선 비대 세포 모델에서 BAE 처리에 의한 IL-6/STAT3 신호 억제 효과 29
• 3.2.1. LPS로 자극된 THP-1 유래 CM(CM-LPS) 처리 염증 자극 모델 확립 29
• 3.2.2. Time-course 분석에 따른 CM-LPS의 전립선 세포 내 IL-6/STAT3 신호전달 조절 33
• 3.2.3. CM 24시간 BAE 동시처리(co-treat) 모델에서 BAE의 IL-6/STAT3 신호전달 억제 효과 39
• 3.2.4. CM 2시간 BAE 동시처리(co-treat) 모델에서 BAE의 IL-6/STAT3 신호전달 억제 효과 43
• 3.2.5. BAE가 전립선 세포 생존율에 미치는 영향 47
• 3.2.6. CM 24시간 BAE 전처리(Pre-treat) 모델에서 BAE의 IL-6/STAT3 신호전달 억제 효과 51
• 3.2.7. CM 2시간 BAE 전처리(Pre-treat) 모델에서 BAE의 IL-6/STAT3축 억제 효과 56
• Ⅳ. 요약 및 결론 60
• 참고문헌 63
• ABSTRACT 72