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课题案例系列之：
铁死亡（Ferroptosis）
案例简介>>
铁死亡是一种由于细胞内代谢通路紊乱导致脂质过氧化物过 度积累引起的细胞死亡方式，与细胞内的铁代谢和脂质稳态 密切相关。
在形态学上，铁死亡与之前发现的细胞凋亡和细胞自噬等存 在明显的差异。铁死亡过程中不会发生细胞破裂的情况。铁 死亡细胞中线粒体膜的密度变大，体积缩小，同时线粒体内 嵴的结构消失，外膜出现破裂。但是铁死亡细胞的细胞核体 积没有明显的变化，核内的染色体结构也不会消失。
|
|
铁死亡 |
凋亡 |
自噬 |
|
特征 |
线粒体嵴消失 |
染色质凝聚， 核仁消失 |
自噬溶酶体形成 |
|
细胞膜系统 |
线粒体外膜破裂，线粒体颜色加深 |
细胞皱缩， 胞质流出胞膜空泡化 |
无变化 |
|
细胞核 |
细胞核不破裂 |
细胞核碎裂， 核仁消失 |
无变化 |
技术路线>>
结果展示>> Fig1
A 
Cell+Cel Cell+LPS Cell + LPS + Drug-A
B C
A. The expression of target molecules was detected by cell immunofluorescence assay, which showed that the expression of target factors significantly decreased after LPS stimulation, and rapidly rebounded under the action of drug A;
B. The Western blot Assay detected the expression of target molecules and showed that the expression of target factors in cells significantly decreased after LPS stimulation, and rapidly rebounded under the action of drug A;
C. The Elisa Assay detected the expression of related inflammatory factors and showed that inflammation increased after cells were stimulated by LPS, and drug A could reverse this process.
技术路线>>
结果展示>> Fig2
A B C D
E
A. RT-qPCR Assay show that after LPS stimulated cells, the expression of key proteins in Ferroptosis related pathways changed. Drug A can reverse this process, which verifies that this model is directly related to Ferroptosis.
B. The key biochemical indicators of Ferroptosis pathways changed after cells were stimulated by LPS. Drug A can reverse this process, which verifies that this model is directly related to Ferroptosis. This experiment detects the expression of this molecule through biochemical Assay;
C. The accumulation of Ferroptosis signaling molecules Fe2+and t-Fe in cells stimulated by LPS can be reversed by drug A, which verifies that this model is directly related to Ferroptosis. This experiment detects the expression of this molecule through biochemical detection.
D. After LPS stimulated cells, the expression of key proteins in Ferroptosis pathways changed. Drug A can reverse this process, which verifies that this model is directly related to Ferroptosis. This experiment use Western Blot assay to detect the expression of molecules.
E. When cells were stimulated by LPS, the mitochondrial crest membrane, which is a marker of Ferroptosis, was ablated or even destroyed. Drug A can reverse this process, verifying that this model is directly related to Ferroptosis. This experiment detected this process through transmission electron microscopy.
技术路线>>
结果展示>> Fig3
A 
B 
C D E
A. After being stimulated by Ferroptosis agonists, cells undergo Apoptosis, and the target protein validation also changes during the Ferroptosis process. Drug A can reverse this process. This experiment detected the expression of this molecule through cellular immunofluorescence Assay;
B. After being stimulated by Ferroptosis agonists, cells undergo Ferroptosis, and the target protein and Ferroptosis related proteins also undergo changes during the Ferroptosis process. Drug A can reverse this process. This experiment detected the expression of related molecules through Western Blot detection;
C. After cells were stimulated by Ferroptosis agonists, iron accumulation occurs in the Ferroptosis related pathways, and the expression of key biochemical indicators changes. Drug A can reverse this process. This experiment is tested through biochemical testing.
D. After cells are stimulated by Ferroptosis agonists, the expression of specific indicators related to Ferroptosis pathways changes, and drug A can reverse this process. This experiment detected the expression of this molecule through Elisa detection method;
E. After being stimulated by Ferroptosis agonists, cells undergo mitochondrial cristae membrane ablation, even destruction, which is a hallmark of Ferroptosis. Drug A can reverse this process. This experiment was validated through transmission electron microscopy detection
技术路线>>
结果展示>> Fig4
A
B C D E
A. After interfering with the target protein, the expression level of the target protein decreased sharply. This experiment detected through cellular immunofluorescence detection;
B. After being stimulated by Ferroptosis agonists, cells undergo Ferroptosis, and the target protein and Ferroptosis related indicators also undergo changes during the Ferroptosis process. Drug A can reverse this process, and after interfering with the target protein, the expression of Ferroptosis related indicators is restored. This experiment detected the expression of this molecule through Western Blot assay;
C. After being stimulated by Ferroptosis agonists, cells undergo Ferroptosis, and drug A can reverse this process. After interfering with the target protein, the expression of biochemical indicators related to Ferroptosis is restored, and the effect of drug A is blocked. This experiment detects through biochemical detection;
D. Drug A can reverse Ferroptosis induced by Ferroptosis inducers, but this process loses its effect with the silencing of the target protein, and the characteristic iron accumulation of Ferroptosis occurs again, blocking the action of drug A. This experiment detects through biochemical detection;
E. Drug A can reverse Ferroptosis induced by Ferroptosis inducers, but this process loses its effect with the silencing of the target protein, and Ferroptosis related indicators are re elevated, and the effect of drug A is blocked. This experiment through biochemical detection;
技术路线>>
结果展示>> Fig5
A B C D
A. Cell ferroptosis can be reversed by overexpression of the target protein, and the blocker of the target protein can block this process, restoring the indicators related to ferroptosis. This experiment detected the inhibition of cell proliferation through CCK8 assay;
B. Cell ferroptosis can be reversed by overexpression of the target protein, and the blocker of the target protein can block this process, restoring the indicators related to ferroptosis. This experiment detected the expression of related proteins in cells through Western Blot;
C. Cell ferroptosis can be reversed by overexpression of the target protein, and the blocker of the target protein can block this process, resulting in the recovery of ferroptosis related protein factors. This experiment detected the expression of related proteins in cells through Elisa's method;
D. Cell ferroptosis can be reversed by overexpression of the target protein, and the blocking agent of the target protein can block this process. The key iron accumulation in ferroptosis is restored, and other related biochemical indicators are corresponding to the table. This experiment detects the accumulation of related factors in cells through biochemical methods;