基于“肾主生殖”理论研究补肾填精法对睾丸氧化应激损伤的保护作用及机制

The theory of “kidney dominating reproduction” is a high-level overview of reproductive physiology. Kidney reinforcing is a basic therapeutic principle to improve male reproduction. Recent research suggests that oxidative stress is an important factor in male infertility, it could damage spermatogenesis, affect steroid hormone synthesis, induce spermatogenic cells apoptosis. Our clinical practice showed that Bushenhuoxue prescription can obviously improve seminal quality, seminal MDA and seminal SOD of oligoasthenospermia patients. We speculate that Bushenhuoxue prescription maybe reduce oxidative stress injury, improve seminal quality, then therapy infertilities with Oligoasthenospermia. While the mechanism is still unclear. In this study, animal experiment and cell experiment were used to investigate the treatment function of Bushenhuoxue prescription on spermatogenic disorder, testosterone synthesis disorder and cells apoptosis induced by oxidative stress. Meanwhile the antioxidant factor Nrf2 and its downstream antioxidant protein heme oxygenase, catalase, superoxide dismutase, glutathione peroxidase expression were analysised to bring light to the mechanism. In conclusion, the study contributes to elaborate the scientific connotation of “kidney dominating reproduction” in Traditional Chinese Medicine, to elucidate its nature and to guide the treatment of male infertility in clinic.

中医“肾主生殖”理论是对生殖生理的高度概括。补肾填精法是改善男性生殖的根本大法。近年研究发现，氧化应激是男性少、弱精症的重要原因，它可以造成精子损伤、睾酮合成障碍、生精细胞凋亡增加等。前期临床研究发现，补肾填精中药补肾活血方可以改善少弱精症患者的精液参数，提高精浆中超氧化物歧化酶的活性，降低过氧化产物丙二醛的水平。课题组推测补肾填精中药可能是通过减少睾丸生精环境的氧化应激损伤，改善精子质量。但其具体机制不清。本项目拟利用氧化损伤动物模型、细胞模型研究补肾活血方对氧化应激引起的生精障碍、睾酮合成障碍、细胞凋亡的保护作用，及对抗氧化核转录因子Nrf2及其下游抗氧化蛋白过氧化氢酶、超氧化物歧化酶、谷胱甘肽过氧化物酶等表达的影响，明确补肾填精中药对睾丸氧化应激损伤的保护作用，阐明其分子机制。这对于深入理解“肾主生殖”的科学内涵，丰富中医药治疗男性不育症的思路以及指导临床，具有重要的理论和现实意义。

中医“肾主生殖”理论是对生殖生理的高度概括。前期临床研究发现，补肾填精中药补肾活血方可以改善少弱精症患者的精液参数，提高精浆中超氧化物歧化酶的活性，降低过氧化产物丙二醛的水平。课题组推测补肾填精中药可能是通过减少睾丸生精环境的氧化应激损伤，改善精子质量。本研究利用环磷酰胺造成小鼠睾丸氧化损伤模型、用过氧化氢造成细胞氧化损伤模型研究补肾活血方对氧化应激引起的生精障碍、睾酮合成障碍、细胞凋亡的保护作用，及对抗氧化核转录因子Nrf2及其下游抗氧化蛋白过氧化氢酶CAT、超氧化物歧化酶SOD2、谷胱甘肽过氧化物酶GPX1等表达的影响，阐明其分子机制。结果表明，补肾活血方可以改善氧化损伤模型小鼠的精子参数、睾丸、附睾的病理损伤情况，增加睾丸组织 SOD、CAT 活性，降低 MDA 含量。补肾活血方可以抑制模型小鼠生精细胞的过度凋亡。补肾活血方可以促进模型鼠睾丸组织中Nrf2-ARE抗氧化信号通路中Nrf2及其下游抗氧化蛋白CAT、SOD2、GPX1 mRNA和蛋白的表达。体外细胞实验表明，补肾活血方可以抑制过氧化氢诱导的精原细胞凋亡、促进其增殖。这表明补肾活血方可以通过对睾丸氧化应激损伤的保护作用来改善生精功能，其机制可能与促进Nrf2-ARE抗氧化信号通路激活，促进抗氧化作用有关。本项目进一步揭示和丰富了“肾藏精，主生殖”的科学内涵，对于指导临床运用中医药治疗男性不育症具有重要的理论意义和临床价值。