Clin Lab. 2022 May 1;68(5). doi: 10.7754/Clin.Lab.2021.211024.
The aim of the study was to evaluate the role of high-mobility group box 1 (HMGB1)-phosphatase and tensin homolog deleted on chromosome ten (PTEN) signaling in T lymphocytes and monocytes upon sepsis.
Thirty C57BL/6 male mice aged 8 - 10 weeks old were randomly divided into sham, model, and inhibitor groups (n = 10). The model of sepsis was established through cecal ligation and perforation. Sham group was only subjected to cecum exposure, and then the wound was sutured without cecal ligation. After the operation, the inhibitor group was intraperitoneally injected with HMGB1-specific inhibitor glycyrrhizic acid (dose: 10 mg/kg) every 6 hours for 4 consecutive times, while sham and model groups were intraperitoneally injected with an equal dose of normal saline. The histopathological changes, cell apoptosis in spleen and thymus tissues, proliferative activity and apoptosis of T lymphocytes, chemotactic activity of monocytes, expression levels of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and IL-10, and expressions of HMGB1 and PTEN in mice were detected using hematoxylineosin staining, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling staining, MTT assay, flow cytometry, transwell chemotaxis assay, enzyme-linked immunosorbent assay, and western blotting, respectively.
The cell apoptosis rate in spleen and thymus tissues, proliferative activity and apoptosis rate of T lymphocytes, chemotactic activity of monocytes, protein expressions of TNF-α, IL-6 and IL-10, and expressions of HMGB1 and PTEN significantly increased in the model group compared with those in the sham group (p < 0.05). However, the cell apoptosis rate in spleen and thymus tissues, T lymphocyte apoptosis rate, protein expressions of TNF-α and IL-6, and expressions of HMGB1 and PTEN were significantly lower, while the proliferative activity of T lymphocytes, chemotactic activity of monocytes and protein expression of IL-10 were significantly higher in the inhibitor group than those in the model group (p < 0.05).
Repressing HMGB1-PTEN signaling can effectively reduce the apoptosis rate of T cells, increase the proliferative activity of T cells, and enhance the function of monocytes in the case of sepsis.
本研究旨在评估高迁移率族蛋白 B1（HMGB1）-磷酸酶和张力蛋白同源物缺失的第十号染色体（PTEN）信号在脓毒症 T 淋巴细胞和单核细胞中的作用。
30 只 8-10 周龄雄性 C57BL/6 小鼠随机分为假手术组、模型组和抑制剂组（每组 10 只）。通过盲肠结扎穿孔术建立脓毒症模型。假手术组仅暴露盲肠，然后缝合切口，不结扎盲肠。手术后，抑制剂组每隔 6 小时腹腔内注射 HMGB1 特异性抑制剂甘草酸（剂量：10mg/kg），连续 4 次，而假手术组和模型组则腹腔内注射等量生理盐水。采用苏木精-伊红染色法、末端脱氧核苷酸转移酶介导的 dUTP 缺口末端标记染色法、MTT 法、流式细胞术、Transwell 趋化实验、酶联免疫吸附试验和 Western blot 法分别检测各组小鼠的组织病理学变化、脾和胸腺组织细胞凋亡、T 淋巴细胞增殖活性和凋亡、单核细胞趋化活性、肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）和白细胞介素-10（IL-10）的表达水平以及 HMGB1 和 PTEN 的表达水平。
与假手术组相比，模型组小鼠脾和胸腺组织细胞凋亡率、T 淋巴细胞增殖活性和凋亡率、单核细胞趋化活性、TNF-α、IL-6 和 IL-10 蛋白表达水平以及 HMGB1 和 PTEN 表达水平均显著升高（p<0.05）。然而，抑制剂组小鼠脾和胸腺组织细胞凋亡率、T 淋巴细胞凋亡率、TNF-α和 IL-6 蛋白表达水平以及 HMGB1 和 PTEN 表达水平均显著降低，T 淋巴细胞增殖活性、单核细胞趋化活性和 IL-10 蛋白表达水平均显著升高（p<0.05）。
抑制 HMGB1-PTEN 信号可有效降低脓毒症时 T 细胞的凋亡率，增加 T 细胞的增殖活性，增强单核细胞的功能。
