康泉方通过BAMBI介导TGF-β/Smad信号通路抑制前列腺增生的机制研究

Kangquan Recipe(KQR), a really curative formula of traditional Chinese medicine(TCM) that was based on the clinical experience and experimental studies of our group over 20 years, has been able to significantly improve the clinical symptoms of BPH patients and suppress hyperplasia of prostate. Our preliminary studys have shown that: KQR has controlled the expression of BAMBI (BMP and activin membrane-bound inhibitor) ’s mRNA levels on BPH-1 cells. Therefore, our research group have put forward a hypothesis like that Kangquan Recipe can regulate the expression of BAMBI which block TGF-β/Smad signaling transduction, thus suppresses the epithelial-mesenchymal transition（EMT）of benign prostate hyperplasia. Then we will test this hypothesis through positive and negative experimental verifications. Firstly, we will explore the different tissues’ expressions of BAMBI on rats treated with KQR, and how the BAMBI blocks the TGF-β/Smad signaling pathways of prostate then to inhibit the EMT of benign prostate hyperplasia. What’s more, we will study the effect of KQR to TGF-β-EMT on BPH-1 cells, and the cells’ BAMBI gene will be silenced. If KQR could not suppress TGF-β-EMT, our hypothesis is established. Finally, mechanism of KQR on BPH will be illuminated via TGF-β-EMT pathways mediated by BAMBI, which will provide a scientific basis for the treatments of KQR on BPH and deepen the research on TCM preventions and therapies for BPH.

康泉方是本课题组经20多年临床与实验研究证明具有确切疗效的经验方，能明显改善良性前列腺增生（BPH）患者临床症状，抑制前列腺增生；前期研究显示：该方能够调控BPH-1细胞中骨形成蛋白和激活素的跨膜抑制剂（BAMBI）等的表达。因此，课题组提出“康泉方通过调控BAMBI表达从而阻断TGF-β/Smad信号通路转导，进而达到抑制前列腺增生组织上皮间质转化(EMT)”的假说，并从正反两方面进行论证：(1)研究康泉方对BPH大鼠不同组织BAMBI的影响，从而竞争性阻断前列腺组织TGF-β/Smad信号通路，减轻前列腺组织EMT现象，达到抑制前列腺增生；(2)研究康泉方在BAMBI沉默的BPH-1细胞系中对TGF-β-EMT通路的作用，如未起到抑制，则反向证明假说成立。阐明该方抑制BPH通过BAMBI介导TGF-β-EMT通路的分子响应机制，为康泉方治疗BPH提供科学依据，深化中医药防治BPH研究。

良性前列腺增生（benign prostatic hyperplasia，BPH）是中老年男性的常见病、多发病。康泉方为本课题组20多年临床实践证明安全有效的经验方。本项目提出“康泉方通过调控BAMBI表达从而阻断TGF-β/Smad信号通路转导，进而达到抑制前列腺增生组织上皮间质转化(EMT)”科学假说，从正反两方面进行论证：(1)研究康泉方对BPH大鼠不同组织BAMBI的影响，从而竞争性阻断前列腺组织TGF-β/Smad信号通路，减轻前列腺组织EMT现象，达到抑制前列腺增生；(2)研究康泉方在BAMBI沉默的BPH-1细胞系中对TGF-β-EMT通路的作用，如未起到抑制作用，则反向证明假说成立。本项目动物实验结果显示，康泉方能够增加大鼠前列腺组织TGF-β、TGF-βR1、TGF-βR2、p-Smad2、p-Smad3、BAMBI、E-cadherin蛋白表达量，减少N-cadherin蛋白表达量；升高前列腺组织TGF-β、TGF-βR1、Smad2、BAMBI、E-cadherin基因表达量，降低N-cadherin基因表达量，说明康泉方能够上调大鼠前列腺等组织BAMBI表达，降低TGF-β生长因子的分泌，阻断TGF-β/Smad信号通路转导，抑制大鼠前列腺增生EMT现象，改善前列腺增生组织和细胞的病理学变化。细胞实验显示转染Si-BAMBI的BPH-1细胞中E-cadherin表达下调，N-cadherin、Vimentin表达上调，未起到抑制EMT作用；更换康泉方含药血清培养BPH-1细胞后，E-cadherin基因的表达上调，N-cadherin、Vimentin表达下调，EMT改善，说明康泉方含药血清通过上调BPH-1细胞中BAMBI的表达，影响TGF/β信号通路相关因子的信号传递，抑制前列腺增生细胞EMT的进程。本项目研究阐明康泉方防治BPH机制的科学内涵，为进一步推广康泉方的临床应用和中药新药研发提供积极的科学依据，造福于BPH病人。