何首乌丸加味干预后衰老模型大鼠睾丸生精细胞p53基因表达的变化  

Changes in p53 expression of spermatogenic cells in testis of aging rats after intervention with modified heshouwu pill

作　　者：陈志宏[1] 龚秀云[1] 李健[2] 薛景凤[1] 

机构地区：[1]承德医学院基础医学研究所,河北省承德市067000 [2]承德市中心医院耳鼻喉科,河北省承德市067000

出　　处：《中国组织工程研究与临床康复》2007年第17期3379-3381,F0003,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：目的:观察中药何首乌丸加味对衰老模型大鼠睾丸生精细胞凋亡、血清睾酮含量和抗氧化能力的影响。方法:实验于2004-04/07在承德医学院基础医学研究所完成。①选用40只雄性SD大鼠,随机分为正常组、模型组、何首乌丸加味组和阴性对照组,每组均为10只。模型组、何首乌丸加味组和阴性对照组均采用D-半乳糖200mg/(kg·d)连续腹腔注射40d制作亚急性衰老大鼠模型。模型组大鼠于造模成功后不再作任何处理;何首乌丸加味用药组大鼠于造模成功后用何首乌丸加味(由何首乌15g,怀牛膝15g,肉苁蓉10g,淫羊藿10g,丹参25g,茯苓15g组成,每副含生药90g,按传统方法加水煎成溶液)8.1g/kg灌胃给药,1次/d,共30d;阴性对照组大鼠于造模成功后每天灌胃同等剂量的生理盐水,时间同何首乌丸加味用药组。正常组大鼠自由摄食、饮水,未加任何干预措施。②所有大鼠经内眦取血2mL,采用酶联免疫吸附法检测各组大鼠血清睾酮含量、黄嘌呤氧化酶法检测血清超氧化物歧化酶的活性;大鼠处死后取睾丸组织、常规石蜡包埋,SP免疫组化法检测睾丸生精细胞p53基因的表达。结果:大鼠40只全部进入结果分析。①血清睾酮含量:模型组明显低于正常组,分别为(0.52±0.14),(1.26±0.32)μg/L(t=3.004,P<0.05);何首乌丸加味组为(1.16±0.32)μg/L,高于模型组(t=2.321,P<0.05)。②血清超氧化物歧化酶活性:模型组明显低于正常组,分别为(106.22±9.63),(148.73±12.93)kNU/L(t=13.339,P<0.01);何首乌丸加味组为(140.05±15.57)kNU/L,高于模型组(t=9.970,P<0.01)。③p53阳性细胞率:模型组明显高于正常组,分别为(59.13±10.53)%和(36.99±9.45)%,(χ2=9.696,P<0.01);何首乌丸加味组为(43.03±12.65)%,低于模型组(χ2=5.122,P<0.05)。结论:何首乌丸加味可通过提高睾酮含量、增强抗氧化能力、降低生精细胞的凋亡改善衰老大鼠睾丸的功能,起到延缓衰老的作用。AIM： To observe the effects of modified heshouwu pill on spermatogenic calls apoptosis in testis, content of serum testosterone and anti-oxidation ability of aging rats. METHODS： The experiment was carded out in the Institute of Basic Medical Sciences of Chengde Medical College from Apdl to July 2004. ①Forty male SD rats were selected and randomly divided into normal group, model group, modified heshouwu pill group and negative control group with 10 rats in each group. The rats except the normal group ware intraperitoneally injected D-gel 200 mg/kg per day continually for 40 days to establish the sub-acute aging rat models. After modeling, no treatment was given to the rats in the model group; rats in the modified heshouwu pill group ware intragastrically perfused of 8.1 g/kg modified heshouwu pill （consisted of 15 g tuber fleecaflower root, 15 g achyranthis root, 10 g desertliving cistanche, 10 g epimedium, 25 g danshen root and 15 g tuckahoe; each dose contained 90 g crude drug, and decocted into solution with water with traditional method）, once a day for 30 days; and those in the negative control group were intragastdcally perfused with saline of the same volume and at the same time. Rats in the normal group took food and drank freely and not received any intervention. ②2 mL blood sample was drawn from the medial canthus of all rats, then the testosterone content in serum was detected by ELISA, the superoxide dismutase （SOD） activity in serum was detected by xanthine oxidase method. After the rats ware killed, the testis tissue was taken out and embedded in paraffin; SP immunohistochemistry was used to observe the p53 expression of spermategenic calls in testis. RESULTS： All the 40 rats ware involved in the result analysis. ①Testosterone content in serum： The model group was obviously lower than the normal group （0.52±0.14）, （1.26±0.32） μg/L, （t =3.004, P〈 0.05）; the content in the modified heshouwu pill group was （1.16±0.32） μg/l_, which was higher than the model

关 键 词：何首乌/药理学 衰老 睾丸 基因 p53 

分 类 号：R286.7[医药卫生—中药学]