Division of General Surgery, Peking University First Hospital, Peking University, 8, Beijing 100034, People's Republic of China; Department of Colorectal Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, People's Republic of China.
Division of General Surgery, Peking University First Hospital, Peking University, 8, Beijing 100034, People's Republic of China.
J Adv Res. 2023 Feb;44:201-212. doi: 10.1016/j.jare.2022.03.010. Epub 2022 Mar 17.
Lipopolysaccharide (LPS) causes lesions of the epithelial barrier, which allows translocation of pathogens from the intestinal lumen to the host's circulation. Hydrogen sulfide (HS) regulates multiple physiological and pathological processes in colonic epithelial tissue, and CBS-HS axis involved in multiple gastrointestinal disorder. However, the mechanism underlying the effect of the CBS-HS axis on the intestinal and systemic inflammation in colitis remains to be illustrated.
To investigate the effect of CBS-HS axis on the intestinal and systematic inflammation related injuries in LPS induced colitis and the underlying mechanisms.
Wild type and CBS mice were used to evaluate the effect of endogenous and exogenous HS on LPS-induced colitis in vivo. Cytokine quantitative antibody array, western blot and real-time PCR were applied to detect the key cytokines in the mechanism of action. Biotin switch of S-sulfhydration, CRISPR/Cas9 mediated knockout, immunofluorescence and ActD chase assay were used in the in vitro experiment to further clarify the molecular mechanisms.
HS significantly alleviated the symptoms of LPS-induced colitis in vivo and attenuated the increase of COX-2 expression. The sulfhydrated HuR increased when CBS express normally or GYY4137 was administered. While after knocking kown CBS, the expression of COX-2 in mice colon increased significantly, and the sulfhydration level of HuR decreased. The results in vitro illustrated that HuR can increase the stability of COX-2 mRNA, and the decrease of COX-2 were due to increased sulfhydration of HuR rather than the reduction of total HuR levels.
These results indicated that CBS-HS axis played an important role in protecting intestinal barrier function in colitis. CBS-HS axis increases the sulfhydration level of HuR, by which reduces the binding of HuR with COX-2 mRNA and inhibited the expression of COX-2.
脂多糖（LPS）可导致上皮屏障损伤，使病原体从肠腔易位至宿主循环。硫化氢（HS）调节结肠上皮组织中的多种生理和病理过程，CBS-HS 轴参与多种胃肠道疾病。然而，CBS-HS 轴对结肠炎中肠道和全身炎症的影响的机制仍有待阐明。
研究 CBS-HS 轴对 LPS 诱导的结肠炎中肠道和全身炎症相关损伤的影响及其潜在机制。
使用野生型和 CBS 小鼠在体内评估内源性和外源性 HS 对 LPS 诱导的结肠炎的影响。采用细胞因子定量抗体阵列、Western blot 和实时 PCR 检测作用机制中的关键细胞因子。采用生物素转换 S-巯基化、CRISPR/Cas9 介导的敲除、免疫荧光和 ActD 追踪实验在体外实验中进一步阐明分子机制。
HS 可显著缓解 LPS 诱导的结肠炎的症状，降低 COX-2 表达的增加。CBS 正常表达或给予 GYY4137 时，HuR 的 S-巯基化增加。而敲除 CBS 后，小鼠结肠中 COX-2 的表达明显增加，HuR 的巯基化水平降低。体外实验结果表明，HuR 可以增加 COX-2 mRNA 的稳定性，而 COX-2 的减少是由于 HuR 发生了增加的巯基化，而不是总 HuR 水平的降低。
这些结果表明，CBS-HS 轴在保护结肠炎中的肠道屏障功能方面发挥了重要作用。CBS-HS 轴增加 HuR 的巯基化水平，通过减少 HuR 与 COX-2 mRNA 的结合，抑制 COX-2 的表达。
