香椿葉萃取物對於盲腸結紮穿刺誘發C57BL/6小鼠敗血症之氧化性傷害的影響評估

Effects of Toona sinensis leaf extracts on the oxidative stress in the C57BL/6 mice with cecal ligation and puncture-induced sepsis

研究背景與目的：敗血症為患者受到感染造成發炎反應並導致組織損傷之疾病，也是住院病患常見的致死病因，先前的研究發現：敗血症的病程發展與氧化壓力增加有關。香椿為一種可以當中藥材服用的植物，研究指出，香椿具有抗氧化的特性，因此本篇研究擬探討香椿葉萃取物對於敗血症小鼠血液及肝組織中在氧化傷害方面的影響。實驗方法：本篇研究以C57BL/6小鼠進行盲腸結紮穿刺手術 (CLP)來誘發敗血症，小鼠分成三組：假手術組 (Sham)、盲腸結紮穿刺組(CLP)、盲腸結紮穿刺補充香椿葉萃取物組(CLP + TS)，手術前先給予小鼠香椿葉萃取物(100mg/kg/day)4週後再進行CLP，分析血液的常規檢查(CBC)、肝功能指標(AST、ALT、ALP)、抗氧化指標(GSH、SOD)；肝組織的抗氧化指標(GSH)、氧化傷害指標(NO)以及與氧化傷害有相關的蛋白質表現(iNOS、HMGB1)。實驗結果：Sham組和CLP組兩者於手術後，相同時間點的比較發現CLP組顯著大於Sham組的有：術後0小時的血液及肝組織GSSG濃度；術後12、15小時的AST及ALT活性，CLP組顯著小於Sham組的有：術後0小時的血液GSH和GSH/GSSG比例；術後3、6、12、15小時的肝組織GSH和total GSH濃度；術後12、15小時的WBC、PLT、單核球比例。CLP組和CLP+TS組兩者於手術後，相同時間點的比較，發現CLP+TS組顯著大於CLP組的有：術後0小時的肝組織GSH濃度；術後15小時的血液GSH、total GSH、GSH/GSSG比例，CLP+TS組顯著小於CLP組的有：術後6小時的iNOS和HMGB1表現量；術後15小時的AST和ALT活性、肝組織NO_2濃度。結論：補充香椿葉萃取物不僅能夠減少肝組織損傷，減少抗氧化物的消耗以及氧化傷害物質的形成，更進一步發現香椿葉萃取物之抗氧化效果與HMGB1的調控有關，可藉由減少iNOS的表現達到控制氧化傷害物質形成的效果。

Background: Sepsis is a leading cause of mortality in hospitals, leading to inflammatory response and tissue injury due to infection. Previous studies have indicated that sepsis is accompanied by increased oxidative stress. The study has discovered that Toona sinensis, a type of traditional Chinese medicine, has an antioxidant effect. Toona sinensis leaf extracts were selected to investigate the effect of oxidative injury on blood and liver tissue in septic mice. Methods: Male C57BL/6 mice were treated with cecal ligation and punctures (CLP) to induce sepsis. Mice were separated into 3 groups: the sham group, CLP group, and CLP+TS group. Toona sinensis leaf extracts (100mg/kg/day) had been pretreated for 4 weeks before CLP. Blood routine (CBC), liver functions (AST, ALT, ALP), antioxidants (SOD, GSH) were included in the analysis of blood. GSH, NO (an oxidative injury indicator), and iNOS and HMGB1 (oxidative injury related proteins) were also included in the analysis of the liver tissue. Results: After CLP (shown as numbers of hours in brackets), comparisons of the sham group and CLP group at the same time point indicated that levels of the blood and liver tissue GSSG (0 hour), and activity of AST and ALT (12, and 15 hours) were significantly higher in the CLP group, whereas levels of the blood GSH and GSH/GSSG ratio (0 hour), levels of the liver tissue GSH and total GSH (3, 6, 12, and 15 hours), levels of the WBC, PLT, and monocyte ratio (12 and 15 hours) were significantly lower in the CLP group. Comparisons of the CLP group and CLP+TS group at the same time point indicated that levels of the liver tissue GSH (0 hour), levels of the blood GSH, total GSH, and GSH/GSSG ratio (15 hours) were significantly higher in the CLP+TS group, whereas expression of iNOS and HMGB1 (6 hours), activity of AST and ALT, level of the liver tissue NO2 (15 hours) were significantly lower in the CLP+TS group. Conclusion: Pretreatment of Toona sinensis leaf extracts could reduce liver tissue injury and oxidative stress, and decrease antioxidant consumption in septic mice. The antioxidant ability of Toona sinensis leaf extracts in septic mice was associated with regulation of the HMGB 1 expression through down-regulation of iNOS to inhibit oxidative stress.