التفاصيل البيبلوغرافية
| العنوان: |
FoxO1 在同型半胱氨酸诱导的肾脏足细胞损伤及 凋亡中的作用研究. (Chinese) |
| العنوان البديل: |
The role of FoxO1 in renal podocyte injury and apoptosis induced by homocysteine. (English) |
| المؤلفون: |
张宏红, 谢琳, 盛思琪, 卢冠军, 姜怡邓 1,2,3，, 杨安宁, 李桂忠 |
| المصدر: |
Tianjin Medical Journal; Jan2022, Vol. 50 Issue 1, p53-58, 6p |
| الملخص (بالإنجليزية): |
Objective To discuss the role of forkhead box O1 (FoxO1) in the injury and apoptosis of podocytes induced by homocysteine (Hcy). Methods Ten Cbs+/+ mice with normal cysteine β -synthase (Cbs) gene and 10 Cbs+/- mice with single gene knockout were fed a high methionine diet. After 8 weeks, the mice were sacrificed to collect kidney tissues. Morphological changes of glomerulus were observed by PAS staining. Podocytes were cultured in vitro and divided into the control group and the Hcy group (treated with cell culture medium containing 80 μmol/L Hcy for 48 h). Podocytes were transfected with Ad-FoxO1 overexpressed adenovirus and Sh-FoxO1 interfering adenovirus carrying green fluorescent protein (GFP), and they were divided into the control group, the Ad-GFP group, the Ad-FoxO1 group, the Sh-NC group, the Sh-FoxO1 group, the Ad-GFP+Hcy group, the Ad-FoxO1+Hcy group, the Sh-NC+Hcy group and the Sh-FoxO1+Hcy group. Western blot assay was used to detect the expression of Podocin and Nephrin, FoxO1 and apoptosis-related B lymphocytoma-2 (Bcl-2), Bcl-2 associated X protein (Bax) and cystine proteinase 12 (Caspase12) proteins in renal tissues and podocytes of mice. The expression of FoxO1 mRNA in mouse kidney tissue and podocytes was detected by qPCR. Results PAS staining results showed that the glomerular structure was normal, the basement membrane was clear and distributed evenly in Cbs+/+ mice, while the glomerular basement membrane presented intermittent thickening and the mesangial matrix increased in Cbs+/- mice. Compared with Cbs+/+ mice, the expression levels of Podocin, Nephrin and FoxO1 protein and FoxO1 mRNA were significantly decreased in Cbs+/- mice (P＜0.01). Compared with the control group, the expression of FoxO1 protein and mRNA in podocytes were significantly decreased in the Hcy group after treated with Hcy (P＜0.01). After overexpression of FoxO1 in podocytes, compared with the Ad-GFP+Hcy group, the protein expression of Podocin and Nephrin were significantly increased, both the ratio of Bax/Bcl-2 and the protein expression of Caspase12 were significantly decreased in the Ad-FoxO1+Hcy group (P＜0.05). After FoxO1 interference, compared with the Sh-NC+Hcy group, Podocin and Nephrin protein expressions were significantly decreased, Bax/Bcl-2 ratio and Caspase12 protein expression were significantly increased in the Sh-FoxO1+Hcy group (P＜0.05). Conclusion FoxO1 can reduce Hcy- induced renal podocyte injury and apoptosis in mice. [ABSTRACT FROM AUTHOR] |
| Abstract (Chinese): |
目的 探讨叉头状转录因子 O1（FoxO1）在同型半胱氨酸（Hcy）诱导的肾脏足细胞损伤及凋亡中的作用。 方法 采用高蛋氨酸饮食喂养胱硫醚 β-合成酶（Cbs）基因正常 Cbs+/+小鼠和单基因敲除 Cbs+/-小鼠各 10 只。8 周后处 死，收集肾组织，PAS 染色观察小鼠肾小球形态学变化。体外培养足细胞，分为 Control 组和 Hcy 组（用含 80 μmol/L Hcy 的细胞培养液干预 48 h）。将携带绿色荧光蛋白（GFP）的过表达 FoxO1 腺病毒（Ad-FoxO1）和干扰 FoxO1 腺病毒 （Sh-FoxO1）转 染 足 细 胞 ，分 为 对 照 组 、Ad-GFP 组 、Ad-FoxO1 组 、Sh-NC 组 、Sh-FoxO1 组 、Ad-GFP+Hcy 组 、Ad- FoxO1+Hcy 组、Sh-NC+Hcy 组、Sh-FoxO1+Hcy 组。Western blot 检测小鼠肾组织和足细胞裂隙膜蛋白（Podocin 和 Nephrin）、FoxO1 及凋亡相关蛋白 B 淋巴细胞瘤-2（Bcl-2）、Bcl-2 相关 X 蛋白（Bax）、半胱氨酸蛋白酶 12（Caspase12） 蛋白表达；qPCR 检测小鼠肾组织和足细胞中 FoxO1 mRNA 的表达。结果 PAS 染色结果显示，Cbs+/+组小鼠肾小球结 构正常，基底膜清晰且分布均匀，而 Cbs+/-组小鼠肾小球基底膜则呈现节段性增厚，系膜基质增多；与 Cbs+/+组小鼠比 较，Cbs+/-组小鼠肾组织中 Podocin、Nephrin 和 FoxO1 蛋白、FoxO1 mRNA 的表达明显降低（P＜0.01）。与 Control 组比 较，Hcy 组 FoxO1 mRNA 和蛋白表达显著降低（P＜0.01）；过表达 FoxO1 后，与 Ad-GFP+Hcy 组相比，Ad-FoxO1+Hcy 组 Podocin 和 Nephrin 蛋白表达均明显升高，Bax/Bcl-2 比值、Caspase12 蛋白表达均明显降低（P＜0.05）；而干扰 FoxO1 后，与 Sh-NC+Hcy 组相比，Sh-FoxO1+Hcy 组 Podocin 和 Nephrin 蛋白表达均明显降低，Bax/Bcl-2 比值、Caspase12 蛋白 表达均明显增高（P＜0.05）。结论 FoxO1 可减轻 Hcy 诱导的小鼠肾脏足细胞损伤及凋亡。 [ABSTRACT FROM AUTHOR] |
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| قاعدة البيانات: |
Complementary Index |
