五子衍宗方对环磷酰胺致小鼠睾丸损伤保护作用-protective effect of wu zi yan zong fang on testicular injury induced by cyclophosphamide in mice.docxVIP

三峡大学硕士学位论文 三 峡 大 学 硕 士 学 位 论 文 PAGE III万方数据 PAGE III 万方数据 Abstract Objective Through intraperitoneal injection CTX to investigate protective effect and mechanism of Wu-Zi-Yan-Zong-Fang against cyclophosphamide induced spermatogenic cells damage in testis in mice. Methods Balb/C mice were randomly divided into five groups: control group, model group, Wu-Zi-Yan-Zong-Fang group with low doses 、 middle and high doses. Wu-Zi-Yan-Zong-Fang groups were given Wu-Zi-Yan-Zong-Fang for three consecutive days prior to the administration of CTX. After 3 days of pretreatment, mice in Wu-Zi-Yan-Zong-Fang groups and model group were administered with CTX (200 mg/kg, i.p.) every week for four times, and mice in the Wu-Zi-Yan-Zong-Fang groups were treated with Wu-Zi-Yan-Zong-Fang. Animals were sacrificed in twelve hours after the final treatment of CTX. The testicular weight, the testis index and Sperm counts, sperm vitality were detected, HE staining was used to observe the histopathology changes; Elisa was used to measure 8-OHdG content in serum; Single cell gel electrophoresis was used to detect DNA of damage in spermatogenic cells in testis; Flow cytometry detection spermatagonial stem cell number; Terminal deoxyribonucleotide transferase mediated dUTP gap markers (Tunel) staining was used to observe the changes of apoptosis; Western blotting was applied to detect the protein expression of P53、p-P53、Bcl-2/bax 、Casepase-3 in spermatogenic cells in testis Results Compared with the model group, Wu-Zi-Yan-Zong-Fang could increased The testicular weight, the testis index, while Sperm counts, sperm vitality were significantly increase (P0.05 or P0.01); HE staining showed that Wu-Zi-Yan-Zong-Fang group can significantly Improve the testicular fine inturn essence exchange function, improve the raw number of sperm cells at various levels; Wu-Zi-Yan-Zong-Fang enhance the anti- DNA damage capability by decreased the level of 8-OHdG, Wu-Zi-Yan-Zong-Fang group