Liu Miaomiao, Zhang Ying, Cao Xiantong, Shi Tao, Yan Yang
Department of Cardiovascular Surgery, First Affiliated Hospital of Xi'an Jiaotong University, Yanta, China.
Cardiol Res Pract. 2022 Feb 17;2022:7687154. doi: 10.1155/2022/7687154. eCollection 2022.
Sepsis is a systemic inflammation and is capable of inducing myocarditis, which is a major leading cause of death in patients. Studies have found that miR-197 is correlated with the prognosis of patients with inflammatory heart disease, but its effect on sepsis-induced cardiomyocyte injury remains unclear. We treated H9c2 cells with lipopolysaccharide (LPS), then detected the cell viability via the cell counting kit-8 (CCK-8) assay and quantified miR-197 expression via quantitative real-time polymerase chain reaction (qRT-PCR). Then, we investigated the role of miR-197 in LPS-induced H9c2 cells by CCK-8 assay, flow cytometry, lactate dehydrogenase (LDH) measurement, enzyme-linked immunosorbent assay (ELISA), qRT-PCR, and western blot. Subsequently, silent information regulator 1 (SIRT1) was downregulated in H9c2 cells to explore its interaction with miR-197 under LPS induction. LPS induced miR-197 overexpression in H9c2 cells. LPS restrained viability, the expressions of B-cell lymphoma-2 (Bcl-2) and SIRT1, but promoted apoptosis, LDH release, and levels of interleukin-6 (IL-6), interleukin-1 (IL-1), acetyl (AC)-p53, BCL2-associated X (Bax), and cleaved caspase-3 in H9c2 cells. miR-197 inhibition reversed the effects of LPS on H9c2 cells. The protective role of miR-197 downregulation in LPS-induced H9c2 cells was reversed by SIRT1 silencing. miR-197 contributed to LPS-induced cardiomyocyte injury by modulating SIRT1, which might be used as a molecular marker in the management of sepsis.
脓毒症是一种全身炎症反应，能够诱发心肌炎，而心肌炎是患者死亡的主要原因。研究发现，miR-197与炎症性心脏病患者的预后相关，但其对脓毒症诱导的心肌细胞损伤的影响仍不清楚。我们用脂多糖（LPS）处理H9c2细胞，然后通过细胞计数试剂盒-8（CCK-8）法检测细胞活力，并通过定量实时聚合酶链反应（qRT-PCR）定量miR-197的表达。然后，我们通过CCK-8法、流式细胞术、乳酸脱氢酶（LDH）测定、酶联免疫吸附测定（ELISA）、qRT-PCR和蛋白质印迹法研究了miR-197在LPS诱导的H9c2细胞中的作用。随后，在H9c2细胞中下调沉默信息调节因子1（SIRT1），以探讨其在LPS诱导下与miR-197的相互作用。LPS诱导H9c2细胞中miR-197表达上调。LPS抑制H9c2细胞的活力、B细胞淋巴瘤-2（Bcl-2）和SIRT1的表达，但促进细胞凋亡、LDH释放以及白细胞介素-6（IL-6）、白细胞介素-1（IL-1）、乙酰化（AC）-p53、BCL2相关X蛋白（Bax）和裂解的半胱天冬酶-3的水平。抑制miR-197可逆转LPS对H9c2细胞的影响。沉默SIRT1可逆转下调miR-197对LPS诱导的H9c2细胞的保护作用。miR-197通过调节SIRT1促进LPS诱导的心肌细胞损伤，这可能作为脓毒症治疗中的一个分子标志物。
