翘芍方重塑早泄大鼠中枢5-HT能系统的机制研究

Premature ejaculation is one of the most common male sexual dysfunction. Serotonin (5-HT) system is an important research target for premature ejaculation.Four pathways of 5-HT synthesis, 5-HT transport, 5-HT receptor(5-HT1A/5-HT2C) and 5-HT metabolic can affect the content and function of 5-HT in synapse.Our previous research results of National Natural Science Foundation of China confirmed that the treatment of premature ejaculation with "Qiao Shao Formula" is safe and effective, and it can imcrease the central 5-HT content of premature ejaculation rats, prolong the Intravaginal ejaculation latency time(IELT).Based on this, we propose that the possible mechanisms of how "QiaoShao Formula" improve the 5-HT content of the central nervous system may be as follows: promoting the synthesis of 5-HT, inhibiting the transport and metabolism of 5-HT, inhibiting the 5-HT1A receptor and stimulating 5-HT2C receptor, and finally remodeling the 5-HT system.The subject adopts the continuum theory of ejaculation to screening model of premature ejaculation. After blocking the above four aspects respectively, then given 4 weeks intervention of "QiaoShao Formula". The detection of each index from the gene, molecular and protein levels to further explore the mechanism of action of "QiaoShao Formula" on premature ejaculation.Then provide a theoretical basis for traditional Chinese medicine in the treatment of premature ejaculation,and provide the reference for clinical treatment of premature ejaculation.

早泄是最常见的男性性功能障碍之一。5-羟色胺（5-HT）能系统是早泄的重要研究靶点，5-HT合成、转运、受体（5-HT1A、5-HT2C）及代谢分解四个环节可影响5-HT在神经突触中的含量及作用。课题组前期国家自然基金项目研究证实，翘芍方治疗早泄安全有效，能够提高早泄大鼠中枢5-HT含量，从而延长射精潜伏期。在此基础上，本课题提出翘芍方提高中枢5-HT含量可能的作用机制如下：促进5-HT的合成，抑制5-HT转运及代谢分解，抑制5-HT1A受体、激动5-HT2C受体，最终重塑了5-HT能系统。课题采用连续统一体理论筛选早泄大鼠模型，分别阻断上述四个环节后，给予翘芍方干预4周，从基因、分子、蛋白水平检测各相关指标，进一步探讨翘芍方治疗早泄的作用机制，为中药治疗早泄提供理论依据，为临床早泄治疗提供参考。

目的：通过动物实验，围绕5-HT系统中5-HT合成、5-HT转运、5-HT代谢分解及5-HT1A受体、5-HT2C受体等进行检测，阐释翘芍方治疗早泄可能作用机制。方法：将60只雄性Wistar大鼠按照随机数字表随机分为空白组、模型组、翘芍方低、中、高剂量组，每组10只。除空白组外，其余5组制备早泄大鼠模型。造模成功后，空白组和模型组采用生理盐水灌胃，翘芍方低、中、高剂量组分别灌胃4周，观察大鼠性行为学，麻醉后取材检测。采用ELISA法检测大鼠下丘脑5-HT、MAOA水平，Real Time PCR检测5-HTT、5-HT1A受体、5-HT2C受体、TPH2mRNA的表达，Western Blot检测下丘脑组织5-HTT、5-HT1A受体、5-HT2C受体、TPH2、蛋白的表达。结果：1 动物性行为学结果显示，与模型组比较，中剂量组ML、EL、MF、EF有显著统计学差异（P < 0.01），IL有统计学差异（P < 0.05）。与模型组比较，高剂量组IL、EL、MF、IF、EF均有显著性差异（P < 0.01）。2 模型组大鼠5-HT下降、MAOA提高，有显著统计学意义（P < 0.01）；翘芍方干预后，可显著提高5-HT含量（P < 0.01），高剂量翘芍方可显著降低MAOA含量（P < 0.01）。3 模型组5-HTT、5-HT1A基因表达升高，TPH2基因表达下降；翘芍方干预后，5-HTT、5-HT1A下降，TPH2表达增强。4 模型组大鼠下丘脑组织中，5-HTT、5-HT1A受体、5-HT2C受体蛋白表达升高，TPH2蛋白表达下降；翘芍方干预后，5-HTT、5-HT1A受体下降，5-HT2C受体、TPH2表达增强。结论：1 翘芍方可改善8-OH-DPAT诱导的早泄大鼠射精过快行为，延长大鼠射精潜伏期。2 翘芍方能够上调TPH2、5-HT2C受体表达，降低5-HTT、5HT1A受体、MAOA表达。通过对5-HT系统中5-HT合成、转运、代谢及受体的综合影响，从而可提高5-HT含量，是其治疗早泄的作用机制之一。