| 摘要: |
| ［摘要］ 目的 探讨精氨酸脱亚胺酶4（PAD4）介导的瓜氨酸化组蛋白H3（H3cit）在脓毒症相关急性肾损伤（AKI）中的作用机制。方法 将30只C57BL/6J小鼠随机分为对照组、模型组和给药组，每组10只。给药组连续7 d灌胃PAD4抑制剂BB-Cl-amidine，其余两组予同体积生理盐水。7 d后对模型组和给药组小鼠腹腔注射脂多糖(LPS)构建AKI模型。24 h后处死小鼠，检测各组小鼠血清肌酐（SCr）、尿素氮（BUN）、H3cit水平，尿液中尿微量白蛋白（UALb）/肌酐（Cr）比值水平，以及肾组织丙二醛（MDA）和超氧化物歧化酶（SOD）水平，并分析SCr、BUN、UALb/Cr比值水平与H3cit水平的相关性。应用HE染色法检测各组小鼠肾脏损伤病理情况。采用Western blot法检测肾脏PAD4和H3cit蛋白表达水平。采用HEK293T细胞进行实验，设置空白组、LPS组、LPS+BB-Cl-amidine组，通过Western blot法检测各组PAD4和H3cit蛋白表达水平，通过CCK-8法检测各组细胞增殖水平。结果 在动物实验中，与对照组比较，模型组小鼠血清SCr、BUN和H3cit水平升高，尿UALb/Cr比值升高，肾组织MDA水平升高、SOD水平降低，肾脏病理损伤加重，PAD4和H3cit蛋白表达升高，差异有统计学意义（P<0.05）。与模型组比较，给药组小鼠血清中SCr、BUN和H3cit水平降低，尿UALb/Cr比值降低，肾组织MDA水平降低、SOD水平升高，肾脏病理损伤缓解，H3cit蛋白表达降低，差异有统计学意义（P<0.05）。H3cit水平与SCr、BUN、UALb/Cr比值水平呈正相关（P<0.05）。在细胞实验中，与空白组比较，LPS组细胞PAD4和H3cit蛋白表达升高，细胞增殖水平降低，差异有统计学意义（P<0.05）。与LPS组比较，LPS+BB-Cl-amidine组细胞H3cit蛋白表达降低，细胞增殖水平升高，差异有统计学意义（P<0.05）。结论 PAD4介导的H3cit可促进脓毒症相关AKI的发展，抑制PAD4活性可以有效减少肾脏细胞损伤。 |
| 关键词: 脓毒症 急性肾损伤 精氨酸脱亚胺酶4 瓜氨酸化组蛋白H3 |
| DOI：10.3969/j.issn.1674-3806.2023.11.12 |
| 分类号:R 692 |
| 基金项目: |
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| A study on the mechanism of action of PAD4-mediated H3cit in sepsis-associated acute kidney injury |
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WEI Ze-feng, WANG Zi-qiang, ZHENG Jin-hua, et al.
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Department of Nephrology, the First Affiliated Hospital of Hainan Medical College, Haikou 570102, China
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| Abstract: |
| ［Abstract］ Objective To study the mechanism of action of peptidylarginine deiminase 4(PAD4)-mediated citrullinated histone H3(H3cit) in sepsis-associated acute kidney injury(AKI). Methods Thirty C57BL/6J mice were randomly divided into control group, model group and drug administration group, with 10 mice in each group. The drug administration group was intragastric administration of PAD4 inhibitor BB-Cl-amidine for 7 days, and the other two groups were given the same volume of normal saline. The AKI model was constructed by intraperitoneal injection of lipopolysaccharide(LPS) in the mice in the model group and the drug administration group 7 days later. The mice were sacrificed 24 hours later, and the levels of serum creatinine(SCr), blood urea nitrogen(BUN) and H3cit, urinary microalbumin(UALb)/creatinine(/Cr) ratio, and the levels of malondialdehyde(MDA) and superoxide dismutase(SOD) in the renal tissues were detected in each group. The correlation of the levels of SCr, BUN and UALb/Cr ratio with H3cit level was analyzed. Hematoxylin and eosin(HE) staining was used to detect the pathological status of kidney injury in mice of each group. The expression levels of PAD4 and H3cit in kidney were detected by Western blot. HEK293T cells were used for the experiment, and the blank group, LPS group and LPS+BB-Cl-amidine group were set up. The expression levels of PAD4 and H3cit protein in each group were detected by Western blot method, and the cell proliferation levels in each group were detected by CCK-8 method. Results In the animal experiments, compared with those in the control group, the levels of serum SCr, BUN and H3cit in the model group were increased, and the urinary UALb/Cr ratio was increased, and the MDA level in the renal tissues was increased, and the SOD level in the renal tissues was decreased, and the renal pathological injury was aggravated, and the expressions of PAD4 and H3cit protein were increased, and the differences were statistically significant(P<0.05). Compared with those in the mice of the model group, the levels of serum SCr, BUN and H3cit were decreased, and the urinary UALb/Cr ratio was decreased, and the MDA level in the renal tissues was decreased, and the SOD level in the renal tissues was increased, and the renal pathological injury was relieved, and the expression of H3cit protein was decreased in the mice of the drug administration group, and the differences were statistically significant(P<0.05). The level of H3cit was positively correlated with the levels of SCr, BUN and UALb/Cr ratio(P<0.05). In the cell experiment, compared with those in the blank group, the expressions of PAD4 and H3cit protein in the LPS group were increased, and the level of cell proliferation in the LPS group was decreased, and the differences were statistically significant(P<0.05). Compared with those in the LPS group, the expression of H3cit protein was decreased and the level of cell proliferation was increased in the LPS+BB-Cl-amidine group, and the differences were statistically significant(P<0.05). Conclusion PAD4-mediated H3cit can promote the development of sepsis-associated AKI, and inhibition of PAD4 activity can effectively reduce kidney cell injury. |
| Key words: Sepsis Acute kidney injury(AKI) Peptidylarginine deiminase 4(PAD4) Citrullinated histone H3(H3cit) |
