Truong Luan D, Trostel Jessica, Roncal Carlos, Cara-Fuentes Gabriel, Miyazaki Makoto, Miyazaki-Anzai Shinobu, Andres-Hernando Ana, Sasai Fumihiko, Lanaspa Miguel, Johnson Richard J, Garcia Gabriela E
Department of Pathology, Baylor College of Medicine, The Houston Methodist Hospital, Houston, Texas.
Division of Renal Diseases and Hypertension, Department of Medicine, University of Colorado Anschutz Medical Campus, Aurora, Colorado.
J Am Soc Nephrol. 2025 Feb 1;36(2):205-218. doi: 10.1681/ASN.0000000000000492. Epub 2024 Sep 20.
Our study demonstrated the sole enzyme responsible for acetylcholine production, choline acetyltransferase, was expressed in podocytes. Acetylcholine decreased glomerular injury in GN by reducing inflammation and protecting endothelium. Choline acetyltransferase/acetylcholine production was induced in podocytes with drugs already available.
One of the most important factors modulating endothelial health is acetylcholine; and while it is associated as a cholinergic neurotransmitter, it is also expressed by non-neuronal cells. However, its role in the kidney, which does not receive cholinergic innervation, remains unknown.
To determine whether acetylcholine is produced in the kidney, we used choline acetyltransferase (ChAT) (BAC)–enhanced green fluorescent protein (ChAT mice) transgenic mice in which enhanced green fluorescent protein is expressed under the control of the endogenous ChAT transcriptional regulatory elements. We then investigated the role of acetylcholine in kidney disease by inducing antiglomerular basement membrane GN (anti-GBM GN) in ChAT transgenic mice.
We demonstrate ChAT, the sole enzyme responsible for acetylcholine production, was expressed in glomerular podocytes and produced acetylcholine. We also show during anti-GBM GN in ChAT transgenic mice, ChAT expression was induced in the glomeruli, mainly in podocytes, and protects mice from kidney injury with marked reduction of glomerular proliferation/fibrinoid necrosis (by 71%), crescent formation (by 98%), and tubular injury (by 78%). By contrast, specific knockout of podocyte ChAT worsened the severity of the disease. The mechanism of protection included reduction of inflammation, attenuation of angiogenic factors reduction, and increase of endothelial nitric oxide synthase expression. and studies demonstrated available drugs such as cholinesterase inhibitors and ChAT inducers increased the expression of podocyte-ChAT and acetylcholine production.
These findings suggest synthesis of acetylcholine by podocytes protected against inflammation and glomerular endothelium damage in anti-GBM GN.
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我们的研究表明，负责乙酰胆碱生成的唯一酶——胆碱乙酰转移酶，在足细胞中表达。乙酰胆碱通过减轻炎症和保护内皮细胞来减轻肾小球肾炎中的肾小球损伤。使用现有药物可诱导足细胞中胆碱乙酰转移酶/乙酰胆碱的生成。
调节内皮细胞健康的最重要因素之一是乙酰胆碱；虽然它作为一种胆碱能神经递质存在关联，但它也由非神经元细胞表达。然而，它在未接受胆碱能神经支配的肾脏中的作用仍不清楚。
为了确定肾脏中是否产生乙酰胆碱，我们使用了胆碱乙酰转移酶（ChAT）（BAC）-增强型绿色荧光蛋白（ChAT小鼠）转基因小鼠，其中增强型绿色荧光蛋白在内源性ChAT转录调控元件的控制下表达。然后，我们通过在ChAT转基因小鼠中诱导抗肾小球基底膜肾小球肾炎（抗GBM GN）来研究乙酰胆碱在肾脏疾病中的作用。
我们证明，负责乙酰胆碱生成的唯一酶ChAT在肾小球足细胞中表达并产生乙酰胆碱。我们还表明，在ChAT转基因小鼠的抗GBM GN期间，肾小球中ChAT表达被诱导，主要在足细胞中，保护小鼠免受肾脏损伤，肾小球增殖/纤维蛋白样坏死显著减少（71%）、新月体形成显著减少（98%）和肾小管损伤显著减少（78%）。相比之下，足细胞ChAT的特异性敲除会加重疾病的严重程度。保护机制包括减轻炎症、减少血管生成因子、增加内皮型一氧化氮合酶表达。 和 研究表明，胆碱酯酶抑制剂和ChAT诱导剂等现有药物可增加足细胞ChAT的表达和乙酰胆碱的生成。
这些发现表明，足细胞合成乙酰胆碱可预防抗GBM GN中的炎症和肾小球内皮细胞损伤。
