Organ Transplantation Center, the First Affiliated Hospital of Kunming Medical University, 650032 Kunming, Yunnan, China.
Gastrointestinal and hernia surgery, the First Affiliated Hospital of Kunming Medical University, 650032 Kunming, Yunnan, China.
Front Biosci (Landmark Ed). 2023 Oct 31;28(10):275. doi: 10.31083/j.fbl2810275.
Heme oxygenase-1 (HO-1) is a protein involved in the inflammatory response following ischemia-reperfusion injury (IRI). Evidence suggests that pyroptosis plays an important role in IRI. However, the underlying mechanism between HO-1 and pyroptosis in IRI requires further investigation.
Using the "two-cuff" method, a Sprague Dawley rat model of liver transplantation (LT) was established using livers from donors after circulatory death. An automatic biochemical analyzer was used to detect serum alanine transaminase (ALT) and aspartate aminotransferase (AST) levels and evaluate liver function. Paraffin sections of the rat liver were stained with hematoxylin-eosin (HE) to observe the degree of pathological damage. An enzyme-linked immunosorbent assay was used to detect serum levels of interleukin (IL)-1β and IL-18. Moreover, western blotting was used to analyze the expression of HO-1, pro-caspase-1, p22, full-gasdermin D (GSDMD), and cleaved-N-GSDMD in the liver. Immunohistochemistry was used to detect NLRP3 expression.
HO-1 expression was time-dependent with IRI. HE staining and Suzuki score showed that necrosis was more severe at 6 h after IRI than in controls. Reactive oxygen species (ROS), ALT, and AST levels in the reperfusion were significantly higher at 6 h after IRI. Similar to HO-1 expression, pro-caspase-1, p22, and GSDMD expression in the reperfusion was time-dependent and was significantly higher at 6 h. Compared with the HO-1-shRNA (short hairpin RNA) group, the HO-1 overexpression group significantly inhibited ROS, p22, GSDMD, IL-1β, IL-18, ALT, and AST. Immunohistochemistry revealed that NLRP3 levels were the highest in the HO-1 overexpression group.
HO-1 improved the survival rate and IRI recovery after LT in rats. This study demonstrates that HO-1 inhibits hepatocyte pyroptosis, thereby reducing IRI after LT.
血红素加氧酶-1（HO-1）是一种参与缺血再灌注损伤（IRI）后炎症反应的蛋白质。有证据表明，细胞焦亡在 IRI 中起着重要作用。然而，HO-1 和 IRI 中细胞焦亡之间的潜在机制需要进一步研究。
使用“双袖套”法，建立了来自心脏死亡供体的肝脏进行肝移植（LT）的 Sprague Dawley 大鼠模型。使用自动生化分析仪检测血清丙氨酸转氨酶（ALT）和天冬氨酸转氨酶（AST）水平，并评估肝功能。对大鼠肝组织进行苏木精-伊红（HE）染色，观察病理损伤程度。酶联免疫吸附试验检测血清白细胞介素（IL）-1β和 IL-18 水平。此外，采用 Western blot 分析肝组织中 HO-1、前半胱天冬酶-1、p22、全长 Gasdermin D（GSDMD）和 cleaved-N-GSDMD 的表达。免疫组化检测 NLRP3 表达。
HO-1 的表达与 IRI 呈时间依赖性。HE 染色和 Suzuki 评分显示，IRI 后 6 小时的坏死程度比对照组更严重。IRI 后 6 小时再灌注中的活性氧（ROS）、ALT 和 AST 水平明显升高。与 HO-1 表达相似，再灌注中的前半胱天冬酶-1、p22 和 GSDMD 的表达也呈时间依赖性，在 6 小时时明显升高。与 HO-1-shRNA（短发夹 RNA）组相比，HO-1 过表达组显著抑制了 ROS、p22、GSDMD、IL-1β、IL-18、ALT 和 AST。免疫组化显示，HO-1 过表达组 NLRP3 水平最高。
HO-1 提高了大鼠 LT 后的存活率和 IRI 恢复。本研究表明，HO-1 抑制了肝细胞焦亡，从而减轻了 LT 后的 IRI。
