Impact of Puberty on the Kidney in Diabetes

青春期对糖尿病肾脏的影响

• 批准号: 6603780
• 负责人: PASCALE H LANE
• 金额: $ 33.08万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6603780
• 关键词: androgen inhibitor; androgen receptor; androgens; angiotensin II; animal puberty; diabetic nephropathy; enzyme linked immunosorbent assay; gender difference; hormone regulation /control mechanism; insulin dependent diabetes mellitus; laboratory rat; male castration; nitric oxide synthase; oxidative stress; pathologic process; protein kinase C; sex hormones; tissue /cell culture; transforming growth factors

DESCRIPTION (provided by applicant): The prepubertal years of type 1 diabetes (DM) appear to be protected from expression of nephropathy and other microvascular complications. Only post-pubertal male rats given the diabetogenic agent streptozocin (STZ) develop renal and glomerular hypertrophy associated with increased expression and activity of transforming growth factor b (TGFb). Prepubertal rats do not develop hypertrophy or upregulation of the TGF system. Given clinical differences in the prevalence and rate of progression between the sexes, gonadal steroids seem likely to be involved in these processes. Overall hypothesis: Androgen synthesis that accompanies puberty contributes to the development of diabetic nephropathy via changes in the renal transforming growth factor (TGF(3) system. Specific Aims: I) What are the roles of androgens in diabetic kidney disease? 1)Examine sex differences in the renal reponse to STZ DM; 2)Examine the effects of gonadectomy on the post-pubertal renal response to STZ DM; 3)Determine the effect of testosterone treatment on the renal response to STZ DM; 4)Determine the role of the androgen receptor in the renal response to DM; and 5)Determine whether conversion to dihydrotestosterone is necessary for the post-pubertal renal response to STZ DM. II) What is the mechanism through which puberty promotes TGFfi expression/activation? 1 )Examine the renin-angiotensin system in response to pre- and post-pubertal states and hormonal manipulation; 2)Examine the protein kinase C system in response to pre- and post-pubertal states and hormonal manipulation; 3)Examine the oxidative stress system in response to pre- and post-pubertal states and hormonal manipulation; 4)Define the direct effects of sex steroids in vitro on the oxidative stress pathway; and 5)Define the direct effects of sex steroids in vitro on the PKC pathway. Methods: Rats will be given STZ DM pre- or post-puberty for 6 weeks, a duration of DM which increases TGFI3 expression and renal weight in adults. Groups will include males and females with and without earlier gonadectomy. Some groups will also receive treatment with testosterone, flutamide, an androgen receptor blocker, or finasteride, which blocks conversion of testosterone to dihydrotestosterone. in vitro studies will involve kidney slice cultures from 10 week old castrated male rats, with or without prior induction of OM. Media will include normal or high glucose conditions, as well as variable amounts of testosterone or estrogen. Measurements will include TGFJ3 proteins by ELISA and nitric oxide synthase isoforms, angiotensin II receptor, and protein kinase C isoforms by immunoblotting; superoxide generation; nitric oxide synthase activity; protein kinase C activity; mRNA for TGFb, nitric oxide synthases, and TGFb inducible gene-H3 by RT-PCR; plasma and renal levels of angiotensin II; and blood levels of sex steroids by RIA. Health implications: New treatments to prevent diabetic kidney disease, the most important cause of kidney failure in the US, may emerge from a better understanding of a naturally protected state such as the prepubertal animal.

描述（由申请人提供）：1型糖尿病的青春期前几年似乎受到保护免受肾病和其他表达的保护 微血管并发症。仅考虑 糖尿病剂链霉菌素（STZ）发展肾脏和肾小球肥大 与转化生长因子的表达增加和活性有关 B（TGFB）。青春期大鼠不会形成肥大或上调 TGF系统。给定临床差异 性别，性腺类固醇之间的进展似乎可能参与 这些过程。总体假设：伴随的雄激素合成 青春 肾转化生长因子（TGF（3）系统。 具体目的：i）雄激素在糖尿病肾脏疾病中的作用是什么？ 1）检查肾脏对STZ DM的性别差异； 2）检查效果 肾上腺切除术对静脉后肾脏对STZ DM的反应； 3）确定 睾丸激素治疗对STZ DM肾脏反应的影响； 4）确定 雄激素受体在肾脏对DM的反应中的作用； 5）确定 在赛后是否需要转换为二氢睾丸激素 对STZ DM的肾脏反应。 ii）青春期的机制是什么 促进TGFFI表达/激活？ 1）检查肾素 - 血管紧张素系统 响应前和后状态和激素操纵； 2）检查蛋白质激酶C系统，以响应前和后伯伯尔 国家和荷尔蒙操纵； 3）检查中的氧化应激系统 对前后状态和荷尔蒙操纵的反应； 4）定义 性类固醇在体外的直接影响对氧化应激途径的直接影响； 5）定义性类固醇在体外对PKC途径的直接影响。 方法：将使大鼠在pe骨前或puberty后6周，持续时间 DM的DM增加了成人TGFI3表达和肾脏重量。小组会 包括有或没有早期性腺切除术的男性和女性。一些小组 还将接受睾丸激素，氟他胺，雄激素受体的治疗 阻断器或非那雄胺，将睾丸激素的转化为阻塞 二氢睾丸激素。体外研究将涉及肾脏切片培养 有或没有事先诱导OM的10周cast割的雄性大鼠。媒体 将包括正常或高葡萄糖条件，以及可变量 睾丸激素或雌激素。测量将包括ELISA和 一氧化氮合酶同工型，血管紧张素II受体和蛋白激酶C 通过免疫印迹的同工型；超氧化物产生；一氧化氮合酶 活动;蛋白激酶C活性； TGFB，一氧化氮合酶和 RT-PCR的TGFB诱导基因-H3；血管紧张素II的血浆和肾脏水平； RIA的性类固醇的血液水平。健康影响：新的治疗方法 预防糖尿病肾脏疾病，这是肾衰竭的最重要原因 美国可能从对自然保护状态的更好理解中脱颖而出 例如前动物。