苦参总黄酮对热应激导致的小鼠生殖功能障碍的治疗作用及机制

Therapeutic Effect and Mechanism of Flavonoids from Sophora Flavescens Ait. on Heat Stress-induced Reproductive Dysfunction in Mice

关键词:  苦参总黄酮  热应激  睾丸组织损伤  TNF-α  IL-17

Key words:  flavonoids from Sophora flavescens Ait.  heat stress  testis tissue injury  TNF-α  IL-17

摘要:

目的 研究苦参总黄酮对阴囊局部热应激引起雄性小鼠睾丸组织损伤的修复作用及机制。方法 使用TCMSP数据库对苦参中黄酮类化合物的靶点进行筛选，对靶点进行生信分析。使用阴囊热应激小鼠模型，苦参黄酮进行干预。检测各组小鼠的精子密度、精子畸变率，观察睾丸组织形态改变。q-PCR和Western blotting检测睾丸肿瘤坏死因子α（tumor necrosis factor-α，TNF-α）和白细胞介素17（interleukin 17，IL-17）的mRNA以及蛋白水平。组织匀浆检测Na⁺-K⁺-ATP酶、Mg²⁺-ATP酶、Ca²⁺-ATP酶、乳酸脱氢酶（lactate dehydrogenase，LDH）、山梨醇脱氢酶（sorbitol dehydrogenase，SDH）活性，丙二醛（malondialdehyde，MDA）、NO、TNF-α的水平及血清中睾酮含量。结果 热应激可以导致小鼠精子的密度减少、畸变的精子增加，睾丸生精上皮明显变薄，细胞层次和数量下降，ATP酶、LDH、SDH的活性显著降低，增加睾丸组织MDA、NO的含量及TNF-α和IL-17的表达。使用250，500 mg·kg^-1·d^-1苦参黄酮干预后，精子质量及睾丸组织形态损伤得到改善，睾丸组织中TNF-α和IL-17的水平降低，ATP酶、SDH活性增高，睾酮水平增高。结论 苦参总黄酮的作用机制是通过抑制炎症因子TNF-α和IL-17的水平，提高抗脂质过氧化能力及抑制NO的作用，增强精子发生的能量酶的活性,提高血清睾酮水平，改善热应激导致的生殖障碍。

Abstract:

OBJECTIVE To investigate the effect and mechanism of flavonoids from Sophora flavescens Ait. on testicular tissue damage in male mice induced by local heat stress in the scrotum. METHODS TCMSP database was used to screen the targets of flavonoids in Sophora flavescens Ait., and the bioinformatics analysis was performed on the target. The mouse model of scrotal heat stress was used and the flavonoids of Sophora flavescens Ait. was used for intervention. The sperm density and sperm aberration rate of mice in each group were measured, and the morphological changes of testicular tissue were observed. Tumor necrosis factor-α (TNF-α) and interleukin 17(IL-17) mRNA and protein levels in testicalar were detected of by q-PCR and Western blotting. Na⁺-K⁺-ATPase, Mg²⁺-ATPase, Ca²⁺-ATPase, lactate dehydrogenase(LDH), sorbitol dehydrogenase(SDH) activity, malondialdehyde (MDA), NO, TNF-α level and the content of testosterone in serum were detected in tissue homogenate. RESULTS Heat stress could lead to the decrease of sperm density and increase of aberrant sperm, the obvious thinning of testicular spermatogenic epithelium, the decrease of cell level and quantity, the significant decrease of ATPase, LDH, SDH activities, and the increase of MDA, NO content, TNF-α and IL-17 expression in testicular tissue. After the intervention with 250, 500 mg·kg^-1·d^-1 flavonoids of Sophora flavescens Ait., the quality of sperm and the damage of testicular tissue morphology were improved. The level of TNF-α and IL-17 in testicular tissue were decreased, and the activity of ATPase, SDH and the level of testosterone were increased. CONCLUSION The mechanism of flavonoids of Sophora flavescens Ait. is through inhibiting the inflammatory factor TNF-α and IL-17 levels, improve the anti-lipid peroxidation ability and inhibite the role of NO, enhance the activity of energy enzymes in spermatogenesis, improve the level of serum testosterone, and improve the reproductive disorders caused by heat stress.