Background

It has been reported that ROCK1 participates in the progression of multiple diseases, including septic intestinal barrier, cardiac dysfunction and acute lung injury. However, its regulatory role and specific mechanism in sepsis-induced acute kidney injury (AKI) remain unclear.

Methods

Cecal ligation puncture (CLP) was conducted to establish sepsis mouse model, and in vitro model was achieved by lipopolysaccharide (LPS) stimulation. Genes expression was evaluated by qRT-PCR, western blot or ELISA was conducted to assess the levels of proteins. Hoechst staining was performed to evaluate cell pyroptosis. LDH activity assay was detected to assess cytotoxicity. Immunohistochemistry was conducted to detect Ly-6G expression and neutrophils distribution in kidney tissues of mice. H&E and TUNEL staining were carried to evaluate kidney injury of mice.

Results

Our findings illuminated that ROCK1 was highly expressed in sepsis-induced AKI, and ROCK1 knockdown inhibited NLRP3-mediated cell pyroptosis in LPS-induced HK-2 cells. Moreover, ROCK1 modulated HK-2 cell pyroptosis by regulating endoplasmic reticulum stress (ERS). TLR2 inhibitor could suppress ERS mediated cell pyroptosis under LPS treatment. Further, TLR2 activator partially reversed the effects of ROCK1 inhibition on ERS mediated pyroptosis in LPS-treated HK-2 cells and CLP mice.

Conclusion

In conclusion, ROCK1 may regulate sepsis-induced AKI via TLR2-mediated ERS/pyroptosis axis. Our data demonstrated the role and underlying mechanism of ROCK1 in septic AKI, providing theoretical basis for sepsis-induced AKI treatment.

中文翻译：

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ROCK1通过TLR2介导的内质网应激/细胞焦亡轴调节败血症诱导的急性肾损伤

背景

据报道，ROCK1 参与多种疾病的进展，包括败血性肠屏障、心脏功能障碍和急性肺损伤。然而，其在败血症诱导的急性肾损伤 (AKI) 中的调节作用和具体机制仍不清楚。

方法

盲肠结扎穿刺（CLP）建立脓毒症小鼠模型，并通过脂多糖（LPS）刺激建立体外模型。通过 qRT-PCR 评估基因表达，进行蛋白质印迹或 ELISA 以评估蛋白质水平。进行Hoechst染色以评估细胞焦亡。检测LDH活性测定以评估细胞毒性。进行免疫组化检测小鼠肾脏组织中Ly-6G的表达和中性粒细胞的分布。进行H＆E和TUNEL染色以评估小鼠的肾损伤。

结果

我们的研究结果表明 ROCK1 在败血症诱导的 AKI 中高度表达，并且 ROCK1 敲低抑制了 LPS 诱导的 HK-2 细胞中 NLRP3 介导的细胞焦亡。此外，ROCK1 通过调节内质网应激 (ERS) 来调节 HK-2 细胞焦亡。TLR2抑制剂在LPS处理下可以抑制ERS介导的细胞焦亡。此外，TLR2 激活剂部分逆转了 ROCK1 抑​​制对 LPS 处理的 HK-2 细胞和 CLP 小鼠中 ERS ​​介导的细胞焦亡的影响。

结论

总之，ROCK1 可能通过 TLR2 介导的 ERS/细胞焦亡轴调节败血症诱导的 AKI。我们的数据证明了 ROCK1 在脓毒症 AKI 中的作用和潜在机制，为脓毒症诱导的 AKI 治疗提供了理论基础。