摘要目的 探讨细胞自噬在去势后大鼠前列腺萎缩中的作用及其机制.方法 将SD大鼠72只随机均分为假切组、去势组、羟氯喹组.各组大鼠分别于术后第1、3、7、10、14、21天随机抽取4只取出前列腺组织.苏木素-伊红(HE)染色观察前列腺组织形态学变化,电镜下观察前列腺细胞自噬小体的变化,免疫组织化学链菌素抗生物素蛋白-过氧化物酶(SP)法检测Beclin1、P62、B淋巴细胞／白血病-2(bcl-2)、bax蛋白在前列腺组织中的表达,逆转录-聚合酶链反应(RT-PCR)检测微管相关蛋白轻链3(LC3)、自噬相关基因5(Atg5) mRNA在前列腺组织中的表达.结果 去势后光镜下大鼠前列腺组织随时间变化逐渐萎缩,电镜下前列腺细胞出现大量空泡状双层膜样结构的自噬小体.去势后Beclin1表达增强(0.0665 ±0.0137),P62表达减弱(0.0103±0.0042),bel-2表达减弱(0.0340±0.0048),bax表达增强(0.0456±0.0122).加用羟氯喹处理后Beclin1表达减弱(0.0103±0.0042),P62表达增强(0.0526±0.0096),bcl-2表达进一步减弱(0.0090±0.0032),bax表达进一步增强(0.0828±0.0098).RT-PCR检测去势后自噬相关基因LC3、Atg5 mRNA表达较假切组增强,用羟氯喹处理后表达减弱.结论 去势后大鼠前列腺细胞自噬增加,羟氯喹可通过阻断自噬促进细胞凋亡,自噬可能通过抑制凋亡而在去势后大鼠前列腺萎缩中起保护作用.

abstractsObjective To investigate the role of autophagy in castration-induced proatate atrophy.Methods Seventy-two healthy male SD rats were randomly divided into 3 groups:sham operation group,castration group and hydroxychloroquine group.At 24 h,72 h,7 days,10 days,14 days and 21 days after operation,we took out four rats randomly from each group and the protaste tissues were removed.The morphological changes of the prostates were observed under a light microscope and the changes of cytolysosomes and apoptotic bodies in the prostates were observed under the electron microscopy.The protein expression of Bechn1,P62,B lymphocytes/leukemia-2 (bcl-2) and bax was detected by detected by using immunohistochemistry.The mRNA expression of microtabule-associated protein 1 light chain 3 (LC3),and autophagy related gene 5 (Atg5) was detected by using reverse transcription-polymerase chain reaction (RT-PCR).Restults The prostate tissues shranked remarkably in the castrated rats.Cytolysosomes were observed under the electron micreseopy.The expressoin of Beelin1 (0.0665 ±0.0137) and bax (0.0456 ±0.0122) was increased,and that of P62 (0.0103 ±0.0042) and bcl-2 (0.0340 ±0.0048) was reduced.After treatment with hydroxychloroquine,the expressoin of Beclin1 (0.0103 ±0.0042) and bcl-2 (0.0090 ±0.0032) was reduced,and that of P62 (0.0526 ±0.0096),and Bax (0.0828±0.0098) was increased.RT-PCR revealed that the mRNA expression of LC3 and Atg5 was enhanced.After treatment with hydroxychloroquine,the mRNA expression of LC3,and Atg5 was weakened.Conclusion The expression of autophagy was increased in prostates after castration.After treatment with hydroxychloroquine,the expression of autophagy was decreased,but apoptosis increased,which suggested that the autophagy can protect the prostates to shinking.