目的 研究引阳索胶囊对腺嘌呤诱导的肾阳虚大鼠精子损伤的保护作用，并从睾丸神经表皮样生长因子2（neuralEGFL like 2，NELL2）Lumicrine信号通路探索其作用机制。方法 雄性SD大鼠随机分为对照组、模型组、引阳索（432mg/kg）组和左卡尼汀（100mg/kg）组，每组10只。连续ig腺嘌呤（200mg/kg）14d建立大鼠肾阳虚模型，并通过观察大鼠肾阳虚证候表现、眼眶取血检测血清中睾酮水平判断造模是否成功；造模同时给予药物干预49d后取材，计算睾丸、附睾脏脑系数；计算机辅助精子分析（computer assisted sperm analysis，CASA）检测附睾精子动力学参数；光镜下进行精子计数、精子畸形率检测；精子荧光检测精子活率；苏木素-伊红（HE）染色观察睾丸、附睾组织病理变化；ELISA 法检测血清中睾酮水平；免疫荧光法观察睾丸NELL2分泌及附睾c-ros癌基因1（ROS proto-oncogene 1，ROS1）表达；Western blotting 检测附睾蛋白酶卵磷脂酶2（ovochymase 2，OVCH2）和精子去整合素金属蛋白酶3（a disintegrin and metalloproteinase 3，ADAM3）表达。结果 与模型组比较，引阳索胶囊可改善肾阳虚大鼠毛发粗糙脱落、精神不振、蜷缩拱背、尿量增多、体质量降低等肾阳虚证候表现，升高睾丸、附睾脏脑系数（P＜0.01）；显著提高精子数量、精子活率、各精子动力学参数（P＜0.05、0.01、0.001），降低精子畸形率，降低曲细精管病变率（P＜0.001），升高14、49d血清中睾酮水平（P＜0.01、0.001）；显著升高睾丸NELL2及附睾ROS1、OVCH2、ADAM3蛋白表达水平（P＜0.05、0.01、0.001）。结论 引阳索胶囊对腺嘌呤诱导的肾阳虚大鼠精子损伤具有保护作用，其机制可能与其调节NELL2 Lumicrine信号通路参与精子发生成熟过程有关。

Objective To study the protective effect of Yinyangsuo Capsules (引阳索胶囊) on sperm damage induced by adenine in rats with kidney-yang deficiency, and explore its mechanism from neural EGFL like 2 (NELL2) Lumirine signal pathway. Methods Male SD rats were randomly divided into control group, model group, Yinyangsuo Capsules (432 mg/kg) group and levocarnitine (100 mg/kg) group with 10 rats in each group. The rat model of kidney-yang deficiency was established by ig adenine (200 mg/kg) for 14 d. The success of model was judged by observing the syndrome of kidney-yang deficiency and testosterone level in serum. At the same time of modeling, drugs were given for 49 d, testicle and epididymis visceral brain coefficients were calculated. Computer assisted sperm analysis (CASA) was used to detect epididymal sperm kinetic parameters. Sperm count and sperm deformity rate were detected under light microscope. Sperm viability was detected by fluorescence; Histopathological changes of testis and epididymis were observed by HE staining. Testosterone level in serum was detected by ELISA. Immunofluorescence was used to observe the secretion of NELL2 in testis and expression of c-ros oncogene 1 (ROS1) in epididymis. The expression of epididymal protease lecithin 2 (OVCH2) and sperm a disintegrin and metalloproteinase 3 (ADAM3) were detected by Western blotting. Results Compared with model group, Yinyangsuo Capsules can improve the symptoms of kidney-yang deficiency in rats with kidney-yang deficiency, such as rough hair loss, listlessness, curled back, increased urine output and decreased body weight, increase the visceral brain coefficient of testis and epididymis (P<0.01); Significantly increased sperm number, sperm viability and sperm dynamics parameters (P<0.05, 0.01, 0.001), decreased sperm deformity rate, decreased seminiferous tubule lesions rate (P<0.001), and increased testosterone level in serum on 14th and 49th d (P<0.01, 0.001); The expression levels of NELL2 in testis and ROS1, OVCH2 and ADAM3 in epididymis were significantly increased (P<0.05, 0.01, 0.001). Conclusion Yinyangsuo Capsules has a protective effect on sperm injury induced by adenine in rats with kidney-yang deficiency, and its mechanism may be related to its regulation of NELL2 Lumicrine signaling pathway to participate in spermatogenesis and maturation.

R285.5

中国中医科学院科技创新工程项目(CI2021A03802);国家自然科学基金资助项目(82004248)