基于氧化及Caspase-3凋亡探讨益肾种子方防治大鼠睾丸损伤机制研究

Study on the Mechanism of Yishen Zhongzi Formula in Preventing and Treating the Testicular Injury of Rats Based on Antioxidation and Caspase-3 Apoptosis

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DOI：10.7661/j.cjim.20230605.241

中文关键词: 益肾种子方睾丸损伤，抗氧化 Caspase-3凋亡途径男性不育症

英文关键词:Yishen Zhongzi Formula testis injury anti-oxidation Caspase-3 apoptosis pathway male infertility

基金项目:国家自然科学基金面上项目（No.81973848）

作者	单位
韩强;曾银;王任远;刘殿池;王和天	首都医科大学附属北京中医医院男科（北京 100010）

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中文摘要:

目的探讨益肾种子方通过抗氧化及其诱导的天冬氨酸特异性半胱氨酸蛋白水解酶（Caspase）-3凋亡途径防治奥硝唑所致的大鼠睾丸损伤的作用机制。方法将30只雄性SD大鼠随机分为5组，空白组、模型组、益肾种子方低、中、高剂量组，每组6只。除空白组外，均采用奥硝唑800 mg/（kg·d）灌胃制造睾丸损伤模型。中药组在造模的同时，分别予益肾种子方低、中、高剂量灌胃[3.37、6.93、13.86 g/（kg·d），每天1次]，连续灌胃28天。干预后取材HE染色观察睾丸及附睾结构变化、测量睾丸组织中活性氧（ROS）、超氧化物歧化酶（SOD）、丙二醛（MDA）、过氧化氢酶（CAT）含量；免疫组织化学染色分析睾丸组织Caspase-3表达。结果与空白组比较，模型组睾丸组织生精细胞不同程度损伤、减少，附睾组织有不同程度破坏和精子的减少，睾丸组织中ROS、MDA含量升高，SOD、CAT含量降低（P<0.01），免疫组化显示Caspase-3表达增多。与模型组比较，益肾种子方干预后睾丸生精细胞完整性有所恢复，Caspase-3明显减少；益肾种子方中剂量组ROS、MDA含量降低（P<0.05）；益肾种子方高剂量组ROS、MDA含量降低（P<0.01），SOD、CAT含量增加（P<0.01，P<0.05）。结论益肾种子方可改善奥硝唑所致大鼠的睾丸及附睾损伤，其机制可能与调控氧化应激系统中ROS、MDA、SOD、CAT等含量，以及氧化应激诱导的Caspase-3凋亡途径有关。

英文摘要:

Objective To investigate the mechanism of Yishen Zhongzi Formula（YZF） in preventing and treating ornidazole-induced testicular injury in rats via anti-oxidation and cysteine-containing aspartatespecific protease（Caspase）-3 apoptosis pathway induced by anti-oxidation. Methods A total of 30 male Sprague-Dawley（SD） rats were randomly divided into 5 groups，blank group，model group，low-，medium- and high-dose YZF group，with 6 rats in each group. The models of testicular injury were established by gavage with 800 mg·kg-1·d-1 ornidazole for all groups except for blank group. In YZF groups，YZF at low，medium and high doses （3.37，6.93 and 13.86 g·kg-1·d-1，respectively，once daily） were administered by gavage for 28 consecutive days during modeling. After intervention，samples were collected for hematoxylin-eosin（HE） staining to observe the structural changes of testis and epididymis，the content of reactive oxygen species（ROS），superoxide dismutase（SOD），malondialdehyde（MDA） and catalase（CAT） in testicular tissues were detected. Moreover，immunohistochemical staining was conducted to analyze the expression of Caspase-3 in testicular tissues. Results Compared with blank group，the spermatogenic cells in testicular tissues were injured and decreased in different degrees，epididymal tissues were damaged in varying degree，sperms were reduced，raised ROS and MDA content but lowered SOD and CAT levels in testicular tissues（P<0.01），and increased Caspase-3 expression was revealed by immunohistochemistry in model group. Compared with model group，the integrity of spermatogenic cells in testicular tissues was restored，and Caspase-3 expression declined significantly following the intervention with YZF. In addition，ROS and MDA content decreased in medium-dose YZF group（P<0.05），reduced ROS and MDA content（P<0.01） and raised SOD and CAT content（P<0.01，P<0.05） were observed in high-dose YZF. Conclusion YZF can ameliorate the ornidazole-induced injury of testis and epididymis in rats，the mechanism may be related to the regulation of ROS，MDA，SOD and CAT content in the oxidative stress system，as well as the Caspase-3 apoptosis pathway induced by oxidative stress.

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