The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Health, Qilu Hospital of Shandong University, Jinan, China; ICU, Anhui Provincial Hospital affiliated Anhui Medical University, Hefei, China.
J Surg Res. 2013 Nov;185(1):286-93. doi: 10.1016/j.jss.2013.05.106. Epub 2013 Jun 24.
The sepsis-induced acute liver injury majorly depends on the dysfunction of mitochondria and the loss of cellular energy. Aquaporin 8 (AQP8) can modulate water transport and osmotic swelling of mitochondria in the inner mitochondrial membrane of the liver. In this study, we explore the effects of tetramethylpyrazine (TMP) on protecting the structure of hepatocyte mitochondria and modulating the expression of AQP8.
Forty-eight rats were randomly allocated to four groups: control group receiving sham procedure, septic group receiving cecal ligation and puncture (CLP), therapeutic group receiving 60 mg/kg of ligustrazine (TMP) intravenously from caudal vein immediately after CLP, and preventive group receiving 60 mg/kg/d of ligustrazine intravenously from caudal vein for 7 d before CLP. The mitochondrial ultrastructure of rat liver was observed. The protein expression of AQP8 was assayed by Western blot. Analysis of AQP8 messenger RNA (mRNA) expression level was performed by the reverse transcription-polymerase chain reaction. The mean fluorescence intensity (MFI) of rhodamine 123 (Rh 123) was measured by flow cytometry. The serum tumor necrosis factor alpha (TNF-α) level was determined by the enzyme-linked immunosorbent assay.
The mitochondrial ultrastructure was markedly damaged in the septic group, whereas it was lightly damaged in the therapeutic and preventive groups. Compared with the control group, the AQP8 protein expression and MFI were significantly reduced, and the steady-state AQP8 mRNA and serum TNF-α levels were increased in the septic, therapeutic, and preventive groups. Compared with the septic group, the AQP8 protein expression and MFI were increased, and the steady-state AQP8 mRNA and serum TNF-α levels were decreased significantly in the therapeutic and preventive groups. There was no significant difference in morphologic characteristics, AQP8 protein level, AQP8 mRNA level, MFI, and serum TNF-α level between the therapeutic and the preventive groups. Linear positive correlation was observed between the AQP8 protein level and the MFI of Rh 123. Linear negative correlation was observed between the AQP8 protein level or the MFI of Rh 123 and serum TNF-α level.
TMP has protective effect on hepatocellular mitochondria from damage in sepsis by ameliorating the expression of AQP8 protein in liver mitochondria. The protective effect of TMP on the liver mitochondria might not have a difference between using TMP before or after the occurrence of sepsis.
脓毒症诱导的急性肝损伤主要取决于线粒体功能障碍和细胞能量丧失。水通道蛋白 8（AQP8）可调节肝线粒体内膜中线粒体的水转运和渗透肿胀。在这项研究中，我们探讨了川芎嗪（TMP）对保护肝细胞线粒体结构和调节 AQP8 表达的作用。
48 只大鼠随机分为 4 组：对照组接受假手术，脓毒症组接受盲肠结扎穿孔（CLP），治疗组在 CLP 后立即从尾静脉给予 60mg/kg 川芎嗪，预防组在 CLP 前 7 天每天从尾静脉给予 60mg/kg/d 川芎嗪。观察大鼠肝线粒体的超微结构。Western blot 法检测 AQP8 蛋白表达。逆转录-聚合酶链反应分析 AQP8 信使 RNA（mRNA）表达水平。通过流式细胞术测量罗丹明 123（Rh 123）的平均荧光强度（MFI）。酶联免疫吸附试验测定血清肿瘤坏死因子-α（TNF-α）水平。
脓毒症组线粒体超微结构明显受损，而治疗组和预防组受损较轻。与对照组相比，脓毒症组、治疗组和预防组 AQP8 蛋白表达和 MFI 均明显降低，稳定态 AQP8mRNA 和血清 TNF-α水平升高。与脓毒症组相比，治疗组和预防组 AQP8 蛋白表达和 MFI 升高，稳定态 AQP8mRNA 和血清 TNF-α水平显著降低。治疗组和预防组之间形态特征、AQP8 蛋白水平、AQP8mRNA 水平、MFI 和血清 TNF-α水平无显著差异。AQP8 蛋白水平与 Rh 123 的 MFI 呈线性正相关。AQP8 蛋白水平或 Rh 123 的 MFI 与血清 TNF-α水平呈线性负相关。
TMP 通过改善肝线粒体 AQP8 蛋白表达对脓毒症引起的肝细胞线粒体损伤具有保护作用。TMP 在脓毒症发生前后使用对肝线粒体的保护作用无差异。
