Department of Physiology and Biophysics, Institute of Biomedical Sciences, University of Sao Paulo, Brazil.
Am J Physiol Regul Integr Comp Physiol. 2011 Jan;300(1):R92-100. doi: 10.1152/ajpregu.00169.2010. Epub 2010 Nov 10.
Endocrine pancreas from pregnant rats undergoes several adaptations that comprise increase in β-cell number, mass and insulin secretion, and reduction of apoptosis. Lactogens are the main hormones that account for these changes. Maternal pancreas, however, returns to a nonpregnant state just after the delivery. The precise mechanism by which this reversal occurs is not settled but, in spite of high lactogen levels, a transient increase in apoptosis was already reported as early as the 3rd day of lactation (L3). Our results revealed that maternal islets displayed a transient increase in DNA fragmentation at L3, in parallel with decreased RAC-alpha serine/threonine-protein kinase (AKT) phosphorylation (pAKT), a known prosurvival kinase. Wortmannin completely abolished the prosurvival action of prolactin (PRL) in cultured islets. Decreased pAKT in L3-islets correlated with increased Tribble 3 (TRB3) expression, a pseudokinase inhibitor of AKT. PERK and eIF2α phosphorylation transiently increased in islets from rats at the first day after delivery, followed by an increase in immunoglobulin heavy chain-binding protein (BiP), activating transcription factor 4 (ATF4), and C/EBP homologous protein (CHOP) in islets from L3 rats. Chromatin immunoprecipitation (ChIP) and Re-ChIP experiments further confirmed increased binding of the heterodimer ATF4/CHOP to the TRB3 promoter in L3 islets. Treatment with PBA, a chemical chaperone that inhibits UPR, restored pAKT levels and inhibited the increase in apoptosis found in L3. Moreover, PBA reduced CHOP and TRB3 levels in β-cell from L3 rats. Altogether, our study collects compelling evidence that UPR underlies the physiological and transient increase in β-cell apoptosis after delivery. The UPR is likely to counteract prosurvival actions of PRL by reducing pAKT through ATF4/CHOP-induced TRB3 expression.
妊娠大鼠的内分泌胰腺经历了几种适应性变化,包括β细胞数量、质量和胰岛素分泌增加,以及细胞凋亡减少。催乳素是导致这些变化的主要激素。然而,母体胰腺在分娩后立即恢复到非妊娠状态。尽管催乳素水平很高,但产后第 3 天(L3)就已经有报道称细胞凋亡短暂增加,确切的逆转机制尚未确定。我们的研究结果表明,母体胰岛在 L3 时显示出 DNA 片段化的短暂增加,同时 RAC-α丝氨酸/苏氨酸蛋白激酶(AKT)磷酸化(pAKT)减少,AKT 是一种已知的促生存激酶。wortmannin 完全消除了催乳素(PRL)在培养胰岛中的促生存作用。L3 胰岛中的 pAKT 减少与 Tribble 3(TRB3)表达增加相关,TRB3 是 AKT 的假激酶抑制剂。产后第 1 天,大鼠胰岛中 PERK 和 eIF2α 磷酸化短暂增加,随后 L3 大鼠胰岛中免疫球蛋白重链结合蛋白(BiP)、激活转录因子 4(ATF4)和 C/EBP 同源蛋白(CHOP)增加。染色质免疫沉淀(ChIP)和再 ChIP 实验进一步证实,ATF4/CHOP 异二聚体在 L3 胰岛中的 TRB3 启动子上的结合增加。用 PBA(一种抑制 UPR 的化学伴侣)处理可恢复 pAKT 水平并抑制 L3 中发现的细胞凋亡增加。此外,PBA 降低了 L3 大鼠胰岛中的 CHOP 和 TRB3 水平。总之,我们的研究提供了令人信服的证据,证明 UPR 是产后β细胞凋亡生理性和短暂增加的基础。UPR 可能通过 ATF4/CHOP 诱导的 TRB3 表达减少 pAKT,从而抵消 PRL 的促生存作用。