2019-08-012024-05-18https://scholars.lib.ntu.edu.tw/handle/123456789/701223摘要：纖維母細胞生長因子23 (Fibroblast growth factor 23, FGF23)，是由骨細胞所分泌的荷爾蒙。其主要的生理功能為(1) 抑制腎臟的磷酸鹽再吸收，降低血磷濃度; (2) 抑制腎臟的活性維生素D3合成以及促進活性維生素D3的分解; (3) 使血中的副甲狀腺素濃度下降。最近許多的證據顯示，在急性腎損傷(acute kidney injury, AKI)的患者身上可以看到有大量的FGF23表現，並發現FGF23可以當作一個急性腎損傷的早期生物標記。臨床的研究結果也顯示，FGF23與急性腎損傷的發生機率有顯著的正相關。另一方面，FGF-23也被證實是腎功能惡化及發生不良預後的預測因子。然而，到目前為止，FGF23促使急性腎損傷的惡化過程，其具體原因仍不清楚，因此需要更多的實驗來證明。為了瞭解上升的FGF23在急性腎損傷的過程中所扮演的角色，我們將FGF23的重組蛋白注射進腎臟缺血再灌流的小鼠中，實驗的結果顯示FGF23重組蛋白可以有效的改善急性腎損傷所造成的傷害。後續的實驗結果顯示，FGF23似乎可以透過活化Akt細胞存活訊息傳遞路徑，在急性腎損傷的小鼠腎臟中，促進腎小管表皮細胞進行大量的再生。另外，FGF23同時也可以減少急性腎損傷的細胞凋亡。這樣令人意外的結果顯示FGF23似乎在急性腎損傷中扮演的是一個保護的作用。 由於我們目前初步的結果只是從單一的實驗動物模式，來探討外源性FGF23與急性腎損傷之間的關聯性。因此，在本研究中，我們將（1）建立三種不同的FGF23功能喪失小鼠之缺血再灌流急性腎損傷動物模式以及另外一種FGF23功能增加小鼠的葉酸注射急性腎損傷動物模式，來提供更多的證據證實FGF23的確在急性腎損傷的過程中是扮演保護的角色。(2) 研究FGF23保護腎臟之分子機制。(3) 找出受到FGF23直接調控之細胞與其中之機制。(4) 在臨床病人上分析FGF23與急性腎損傷之間的關聯性。本研究計畫的成果將會打破我們以往對於FGF23與急性腎損傷之間的認知。<br> Abstract: Fibroblast growth factor 23 (FGF23) is a hormone which is predominately produced in bone. The main physiological functions of FGF23 are to prevent renal phosphate reabsorption, active vitamin D3 production by the kidney, and decrease the parathyroid hormone (PTH) concentration in the blood. Recent evidences indicated that FGF23 rapidly increases after acute kidney injury (AKI), and can serve as a biomarker for subsequent AKI as well as adverse outcomes post-AKI. However, the biologic role for this finding is still unclear and requires further investigation.To explore the effect of increased FGF23 during AKI, we injected FGF23 recombinant protein into ischemia-reperfusion induced acute kidney injury (IR-AKI) mice. The preliminary results show that FGF23 could ameliorates kidney injury which induced by ischemia-reperfusion injury. Western blot results showed that activated Akt survival pathway was much greater in the kidney of FGF23-treated IR-AKI group than that in IR-AKI group. Proliferation marker PCNA and differentiation marker E-cadherin were also upregulated in IR-AKI mouse kidney after FGF23 pretreated. Activated Akt and PCNA overexpression were found in renal tubular epithelium by immunohistochemistry analysis. These results showed that FGF23 accelerates the process of tubular regeneration. Furthermore, FGF23 could prevent IR-induced cell death in the AKI mouse kidney by TUNEL assay. Taken together, our incredible results suggested that FGF23 seems to play a critical role in protection from IR-AKI.However, our preliminary results were obtained from a single exogenous FGF23-treated mouse model. Therefore, in this project, we will shed light on (1) The protective role of FGF23 in AKI using another three different FGF23 loss-of-function animal models and one FGF23 gain-of-function folic acid-induced AKI model. (2) The molecular mechanism underlying the protection of renal tubules by FGF23 using two in vivo models. (3) The identification of critical cell types and molecular mechanisms directly involved in the protection against AKI by FGF23. (4) The association between FGF23 and the prognosis of AKI in critical patients. This project will lead to breakthrough in our understanding of the link between FGF23 and AKI.纖維母細胞生長因子23 (Fibroblast growth factor 23, FGF23)，是由骨細胞所分泌的荷爾蒙。其主要的生理功能為(1) 抑制腎臟的磷酸鹽再吸收，降低血磷濃度; (2) 抑制腎臟的活性維生素D3合成以及促進活性維生素D3的分解; (3) 使血中的副甲狀腺素濃度下降。最近許多的證據顯示，在急性腎損傷(acute kidney injury, AKI)的患者身上可以看到有大量的FGF23表現，並發現FGF23可以當作一個急性腎損傷的早期生物標記。臨床的研究結果也顯示，FGF23與急性腎損傷的發生機率有顯著的正相關。另一方面，FGF-23也被證實是腎功能惡化及發生不良預後的預測因子。然而，到目前為止，FGF23促使急性腎損傷的惡化過程，其具體原因仍不清楚，因此需要更多的實驗來證明。 為了瞭解上升的FGF23在急性腎損傷的過程中所扮演的角色，我們將FGF23的重組蛋白注射進腎臟缺血再灌流的小鼠中，實驗的結果顯示FGF23重組蛋白可以有效的改善急性腎損傷所造成的傷害。後續的實驗結果顯示，FGF23似乎可以透過活化Akt細胞存活訊息傳遞路徑，在急性腎損傷的小鼠腎臟中，促進腎小管表皮細胞進行大量的再生。另外，FGF23同時也可以減少急性腎損傷的細胞凋亡。這樣令人意外的結果顯示FGF23似乎在急性腎損傷中扮演的是一個保護的作用。 由於我們目前初步的結果只是從單一的實驗動物模式，來探討外源性FGF23與急性腎損傷之間的關聯性。因此，在本研究中，我們將（1）建立三種不同的FGF23功能喪失小鼠之缺血再灌流急性腎損傷動物模式以及另外一種FGF23功能增加小鼠的葉酸注射急性腎損傷動物模式，來提供更多的證據證實FGF23的確在急性腎損傷的過程中是扮演保護的角色。(2) 研究FGF23保護腎臟之分子機制。(3) 找出受到FGF23直接調控之細胞與其中之機制。(4) 在臨床病人上分析FGF23與急性腎損傷之間的關聯性。本研究計畫的成果將會打破我們以往對於FGF23與急性腎損傷之間的認知。Fibroblast growth factor 23 (FGF23) is a hormone which is predominately produced in bone. The main physiological functions of FGF23 are to prevent renal phosphate reabsorption, active vitamin D3 production by the kidney, and decrease the parathyroid hormone (PTH) concentration in the blood. Recent evidences indicated that FGF23 rapidly increases after acute kidney injury (AKI), and can serve as a biomarker for subsequent AKI as well as adverse outcomes post-AKI. However, the biologic role for this finding is still unclear and requires further investigation. To explore the effect of increased FGF23 during AKI, we injected FGF23 recombinant protein into ischemia-reperfusion induced acute kidney injury (IR-AKI) mice. The preliminary results show that FGF23 could ameliorates kidney injury which induced by ischemia-reperfusion injury. Western blot results showed that activated Akt survival pathway was much greater in the kidney of FGF23-treated IR-AKI group than that in IR-AKI group. Proliferation marker PCNA and differentiation marker E-cadherin were also upregulated in IR-AKI mouse kidney after FGF23 pretreated. Activated Akt and PCNA overexpression were found in renal tubular epithelium by immunohistochemistry analysis. These results showed that FGF23 accelerates the process of tubular regeneration. Furthermore, FGF23 could prevent IR-induced cell death in the AKI mouse kidney by TUNEL assay. Taken together, our incredible results suggested that FGF23 seems to play a critical role in protection from IR-AKI. However, our preliminary results were obtained from a single exogenous FGF23-treated mouse model. Therefore, in this project, we will shed light on (1) The protective role of FGF23 in AKI using another three different FGF23 loss-of-function animal models and one FGF23 gain-of-function folic acid-induced AKI model. (2) The molecular mechanism underlying the protection of renal tubules by FGF23 using two in vivo models. (3) The identification of critical cell types and molecular mechanisms directly involved in the protection against AKI by FGF23. (4) The association between FGF23 and the prognosis of AKI in critical patients. This project will lead to breakthrough in our understanding of the link between FGF23 and AKI.急性腎損傷纖維母細胞生長因子23缺血/再灌注損傷Acute kidney injuryFibroblast growth factor 23Ischemia-reperfusion injury