淫羊藿苷对双酚A和肾阳虚诱导睾丸雄激素受体磷酸化影响的比较研究  

Comparative Study on the Effect of Icariin on Androgen Receptor Phosphorylation in Testis Induced by Bisphenol A and Shen-Yang Deficiency

作　　者：张学学 马静 贾瑊 许航 崔梁瑜 ZHANG Xue-xue;MA Jing;JIA Jian;XU Hang;CUI Liang-yu(Department of Traditional Chinese Medicine,Xijing Hospital of the Air Force Military Medical University,Xi’an 710032;Graduate School,Shaanxi University of Chinese Medicine,Shaanxi 712046)

机构地区：[1]空军军医大学西京医院中医科,西安710032 [2]陕西中医药大学研究生院,陕西712046

出　　处：《中国中西医结合杂志》2022年第6期733-739,共7页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.81573851);陕西省国际科技合作与交流计划(No.2016KW-012)。

摘　　要：目的观察淫羊藿苷(ICA)对双酚A(BPA)染毒和肾阳虚小鼠睾丸细胞凋亡及雄激素受体(AR)磷酸化影响,探讨ICA作用机制。方法45只SPF级雄性昆明小鼠随机分为正常组(0.2 mL/d生理盐水)、BPA组(0.2 mL/d生理盐水)、BPA+ICA低、中、高剂量组[50、100、200 mg/(kg·d)ICA]及肾阳虚组(0.2 mL/d生理盐水)、肾阳虚+ICA低、中、高剂量组[50、100、200 mg/(kg·d)ICA],每组5只。BPA各组采用腹腔注射100 mg/(kg·d)BPA连续7 d制备模型,肾阳虚各组采用腹腔注射氢化可的松注射液25 mg/(kg·d)连续10 d制备模型,各组小鼠造模后灌胃相应药物14 d。采用ELISA法检测血清T、E2水平,TUNEL染色观察睾丸细胞凋亡,免疫组化染色、免疫荧光染色及Western Blot法观察睾丸组织AR、磷酸化AR(p-AR)、血小板丝氨酸/苏氨酸蛋白激酶(AKT)及磷酸化AKT(p-AKT)的表达。结果与正常组比较,BPA组和肾阳虚组小鼠睾丸结构出现不同程度损伤,细胞凋亡、血清E2水平、睾丸p-AKT、p-AR表达增加,血清T水平、T/E2、睾丸AR表达下降(P<0.05,P<0.01)。与BPA组比较,BPA+低剂量组T/E2升高,睾丸p-AR、p-AKT表达降低(P<0.05,P<0.01);BPA+ICA中剂量组小鼠睾丸组织结构改善,细胞凋亡、血清E2水平、睾丸p-AKT、p-AR表达降低,血清T水平、T/E2、睾丸AR表达升高(P<0.05,P<0.01);BPA+ICA高剂量组细胞凋亡、血清T水平、睾丸AR表达增加,睾丸p-AR、p-AKT表达降低(P<0.05,P<0.01)。与肾阳虚组比较,肾阳虚+ICA低剂量组血清E2水平、睾丸p-AR、p-AKT表达下降,睾丸AR表达增加(P<0.05,P<0.01);肾阳虚+ICA中剂量组小鼠睾丸组织结构改善,细胞凋亡、E2水平、睾丸p-AR、p-AKT表达降低,血清T水平、T/E2、睾丸AR表达增加(P<0.05,P<0.01);肾阳虚+ICA高剂量组细胞凋亡、血清T水平、T/E2、睾丸AR表达增加,睾丸p-AR、p-AKT表达降低(P<0.05,P<0.01)。结论ICA可以抑制BPA和肾阳虚诱导睾丸细胞凋亡,机制与上调AR表达、下调p-AR及p-AKT表达有关,�Objective To observe the effects of icariin(ICA)on testicular cell apoptosis and androgen receptor(AR)phosphorylation in bisphenol A(BPA)poisoning and Shen-Yang deficiency(SYD)mice,and to explore the mechanism.Methods Totally 45 SPF male Kunming mice were randomly divided into normal control group(0.2 mL/d normal saline),BPA group(0.2 mL/d normal saline),BPA+ICA low-dose,medium-dose and high-dose group[BPA+50,100,200 mg/(kg·d)ICA],SYD group(0.2 mL/d normal saline),SYD+ICA lowdose,medium-dose and high-dose group[SYD+50,100,200 mg/(kg·d)ICA],5 mice in each group.BPA groups was intraperitoneally injected 100 mg/(kg·d)BPA solution for 7 consecutive days,KYD groups received intraperitoneal injected 25 mg/(kg·d)hydrocortisone for 10 consecutive days to build model,mice in each group were intragastric with corresponding drugs for 14 days after modeling.Serum testosterone(T)and estradiol(E2)levels were detected by ELISA.Apoptosis of testicular cell was observed by TUNEL staining.AR,phospho-AR(p-AR),serine/threonine-protein kinase(AKT)and phospho-AKT(p-AKT)in testicular tissue were detected by immunohistochemical staining,immunofluorescence staining and Western Blot.Results Compared with normal group,testicular structure showed different degrees of damage,cell apoptosis,serum E2 level,testicular p-AKT and p-AR expression increased,serum T level,T/E2 and testicular AR expression decreased in BPA group and KYD group(P<0.05,P<0.01).Compared with BPA group,T/E2 increased,testis p-AR and P-AKT expression decreased in BPA+ICA low-dose group(P<0.05,P<0.01).Testicular tissue structure was improved,cell apoptosis,serum E2 level,testicular p-Akt and P-AR expression decreased,serum T level,T/E2 and testicular AR expression increased in BPA+ICA medium-dose group(P<0.05,P<0.01).Cell apoptosis,serum T level and testicular AR expression increased,testicular p-AR and p-AKT expression decreased in BPA+ICA high-dose group(P<0.05,P<0.01).Compared with KYD group,serum E2 level,testicular p-AR and P-AKT expression decreased,testicular AR

关 键 词：淫羊藿苷 双酚A 肾阳虚 睾丸 细胞凋亡 雄激素受体磷酸化 

分 类 号：R285.5[医药卫生—中药学]