Heat stress triggers cell death through activating RIPK3 in vitro
热应激在体内通过激活RIPK3触发细胞死亡
We examined whether heat stress alone could activate RIPK3-dependent cell death.
我们测试了热应激能否独立激活RIPK3依赖性的细胞死亡
Exposure of cultured L929 mouse fibroblasts to 43°C for 2 hours or 42°C for 6 hours induced the phosphorylation of RIPK3 and MLKL within 2 hours and the cleavage of pro-casp8, pro-casp3, and GSDME by 6 hours after heat stressexposure (Fig. 2A).
将L929小鼠成纤维细胞置于43摄氏度2小时或42摄氏度6小时环境培养，在高温培养后，可诱导RIPK3和MLKL2小时内磷酸化，前casp-8蛋白，前casp-9蛋白和GSDME在六小时后裂解
Deletion of RIPK3 using CRISPR-CAS9 blocked heat stress–induced phosphorylation of RIPK3 and MLKL and the cleavage of pro-casp8, pro-casp3, and GSDME(Fig. 2A).
运用CRISPR-CAS9将RIPK3敲除，可阻断热应激诱导的RIPK3和MLKL的磷酸化，前casp-8蛋白，前casp-9蛋白和GSDME的裂解
These events were associated with a reduction of cell death as judged by lactate dehydrogenase (LDH) release, loss of cell membrane integrity, and increased cell membrane permeability .
这些现象同时伴随着细胞死亡的减少，可根据LDH释放量，细胞膜完整度和通透性加以判断
Heat stress promoted the interaction between RIPK3 and MLKL in WT but not Ripk3-deficient L929 cells (fig. S3, D and E).
在WT细胞中，热应激会促进RIPK3和MLKL的相互作用，但RIPK-3缺失的L929细胞中不会出现
Depletion of RIPK3 with short hairpin RNA (shRNA) inhibited heat stress–induced cell death (fig. S3F),whereas reconstitution of the expression of RIPK3 restored the capacity of Ripk3-deficient L929 cells to undergo cell death in response to heat stress (Fig. 2D).
用shRNA沉默RIPK3抑制了热应激诱导的细胞死亡，而RIPK3的表达的重建恢复了RIPK3缺陷的L929细胞的响应热应激激活细胞死亡的能力
Similar observations were made using bone marrow–derived macrophages (BMDMs) and peritoneal macrophages(PMs) from WT or Ripk3−/− mice .
WT小鼠和RIPK3敲除小鼠的骨髓来源的巨噬细胞和腹腔巨噬细胞中也有类似的发现
Although MLKL deficiency protected cells from heat stress–induced necroptosis within 12 hours, delayed cell death occurred 24 hours after heat stress exposure in Mlkl−/− but not Ripk3−/− BMDMs (Fig. 2E).
尽管MLKL的缺失保护细胞在12h内免于热应激诱导的坏死，但在mlkl缺失的细胞热应激24h后还是会发生细胞死亡，而在RIPK3缺失BMDM细胞中不会发生
Because RIPK3 can mediate cell death through its scaffolding function and casp8 (15), we thought heat stress might induce RIPK3-dependent cell death through both MLKL and casp8.
因为RIPK3可以通过其支架功能和casp8调控细胞死亡，我们认为热应激诱导RIPK3依赖性的细胞死亡通过MLKL和casp8 两个方式实现。
今天起得很晚，又啥也没学
再也不可以了！！！
