Kisspeptin神经元网络：电针调节青春期PCOS神经内分泌异常的靶点环节？

The onset of polycystic ovarian syndrome (PCOS) is usually in adolescence. Adolescence is a phase during which hypothalamus-pituitary-ovary axis (HPOA) is developing and full of strong plasticity. Therefore, it will delay, inhibit, or even reverse the development of PCOS if we treat PCOS as early as in adolescence. The characteristic neuroendocrine abnormality of adolescent PCOS is over-stimulation of GnRH neuron in hypothalamus. Kisspeptin neuron is the key intermedia to relay all kinds of signals to GnRH neuron. This project speculate that abnormal onset of adolescence with too strong excitatory signals and/or too weak inhibitory signals which makes too much secretion and/or too strong effect of kisspeptin, is the important mechanism of GnRH neuron over-stimulation in adolescent PCOS; electro-acupuncture (EA), as a kind of stimulatory signal, can inhibit the over-stimulation of GnRH neuron through regulating kisspeptin neuron network, and then improve neuroendocrine abnormality of adolescent PCOS. Utilizing techniques of real-time RT-PCR, western blot, in situ hybridization and double-labbed immunofluorescence combined with confocal microscopy, the project will probe the mechanisms of EA in regulating neuroendocrine abnormality in adolescent PCOS from the target point of kisspeptin neuron network including presynaptic and postsynaptic signaling pathways.

PCOS多起病于青春期，此时HPOA具有很强的可塑性，及早在青春期对PCOS进行干预，可延缓、抑制甚或逆转其发展。青春期PCOS典型的神经内分泌异常就是下丘脑GnRH神经元的过度激活。Kisspeptin神经元是机体将各种信号传递给GnRH神经元的关键媒介。本项目推测，青春期启动异常，兴奋性信号过度增强和/或抑制性信号过度减弱，使得Kisspeptin分泌增多或作用增强，是导致青春期PCOS的GnRH神经元过度激活的重要机制；电针作为一种刺激信号，通过调节Kisspeptin神经元网络抑制GnRH神经元的过度激活，从而改善青春期PCOS的神经内分泌异常。本项目采用实时定量RT-PCR技术、Western blot法、原位杂交技术、免疫荧光双标记结合激光共聚焦显微技术，以Kisspeptin神经元网络（突触前和突触后信号通路两方面）为靶点，探讨电针调节青春期PCOS神经内分泌异常的作用机制。

本项目建立青春期PCOS大鼠模型，探讨了电针对青春期 PCOS 模型大鼠动情周期、内分泌和糖脂代谢异常的调节作用；并从kisspeptin神经元网络（包括Kisspeptin/dynorphin/neurokinin B/GnRH，以及Kisspeptin神经元突触前和后信号通路）探讨了其作用的分子机制。本项目研究明确了电针对青春期PCOS大鼠动情周期、卵巢形态学、性激素紊乱以及糖脂代谢紊乱的改善作用。本研究结果显示其作用机制可能包括以下几点：（1）电针可下调青春期PCOS大鼠下丘脑中Kisspeptin表达，上调GnRH表达。（2）电针使青春期PCOS大鼠血清中ghrelin水平升高，使下丘脑中ghrelin蛋白和mRNA表达增加，下丘脑弓状核中GHRL蛋白表达升高。（3）电针下调青春期PCOS大鼠下丘脑中NPY的mRNA水平，电针使青春期PCOS大鼠下丘脑弓状核中NPY蛋白表达下降，电针明显增加下丘脑NPY2R的蛋白和mRNA水平。本研究为电针早期干预青春期PCOS提供了一定的实验依据。