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課題案例系列之：
鐵死亡（Ferroptosis）
案例簡介>>
鐵死亡是一種由于細(xì)胞內(nèi)代謝通路紊亂導(dǎo)致脂質(zhì)過氧化物過 度積累引起的細(xì)胞死亡方式，與細(xì)胞內(nèi)的鐵代謝和脂質(zhì)穩(wěn)態(tài) 密切相關(guān)。
在形態(tài)學(xué)上，鐵死亡與之前發(fā)現(xiàn)的細(xì)胞凋亡和細(xì)胞自噬等存 在明顯的差異。鐵死亡過程中不會(huì)發(fā)生細(xì)胞破裂的情況。鐵 死亡細(xì)胞中線粒體膜的密度變大，體積縮小，同時(shí)線粒體內(nèi) 嵴的結(jié)構(gòu)消失，外膜出現(xiàn)破裂。但是鐵死亡細(xì)胞的細(xì)胞核體 積沒有明顯的變化，核內(nèi)的染色體結(jié)構(gòu)也不會(huì)消失。
|
|
鐵死亡 |
凋亡 |
自噬 |
|
特征 |
線粒體嵴消失 |
染色質(zhì)凝聚， 核仁消失 |
自噬溶酶體形成 |
|
細(xì)胞膜系統(tǒng) |
線粒體外膜破裂，線粒體顏色加深 |
細(xì)胞皺縮， 胞質(zhì)流出胞膜空泡化 |
無變化 |
|
細(xì)胞核 |
細(xì)胞核不破裂 |
細(xì)胞核碎裂， 核仁消失 |
無變化 |
技術(shù)路線>>
結(jié)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.
技術(shù)路線>>
結(jié)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.
???技術(shù)路線>>
結(jié)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
技術(shù)路線>>
結(jié)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;
技術(shù)路線>>
結(jié)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;