添加精胺酸對腹膜炎引致敗血症老鼠腸道及全身性免疫反應之影響

本實驗探討在敗血症前後添加精胺酸（Arginine, Arg）對敗血症時營養素代謝及腸道免疫反應之影響。實驗以Wistar雄性老鼠為實驗對象，共分為四組，第一、二組給予一般飼料組成之飲食，第三、四組給予以總熱量2% Arg取代部分酪蛋白之飲食，兩組為等氮量，各給予十天後，進行盲腸結紮及穿刺手術（cecal ligation and puncture）以引致敗血症，同時進行頸靜脈插管手術，第一、三組老鼠輸入一般之TPN輸液，第二、四組輸入含2% Arg之營養輸液，四組分別為第一組：敗血致前後均無補充Arg組。第二組：敗血症後給予Arg補充組。第三組：敗血症前給予Arg補充組。第四組：敗血症前後均補充Arg組。在引致敗血症後第三天犧牲老鼠，取血液、腹腔灌洗液、脾臟及小腸做分析。結果顯示在營養素代謝方面，Arg之補充對於血糖並無影響，但第三組及第四組其血中三酸甘油酯及游離脂肪酸濃度明顯較第一組及第二組低，顯示在敗血症前補充Arg可使脂肪組織分解的情形獲得改善；在蛋白質代謝方面，Arg之補充可使血中Arg濃度顯著增高，但支鏈胺基酸和glutamine之濃度，在四組間並無顯著之差異，顯示補充Arg對肌肉組織異化作用影響並不明顯。在免疫反應方面，以第三組、第四組與第一組、第二組比較，其腹腔灌洗液中好氧菌或厭氧菌之總菌落數明顯降低，而巨噬細胞之吞噬能力明顯較強；第四組小腸上Peyer’s patches之總顆粒數和其中淋巴細胞數量、小腸灌洗液中total immunoglobulin A濃度，以及脾臟細胞經刺激後產生Interleukin-10之濃度均明顯高於第一組及第二組；不論添加的時間，補充Arg的3組其Interferon-γ濃度明顯較第四組為低。以上之結果顯示，在敗血症前補充Arg之組別可促進發炎局部之體液性及非特異性免疫反應，但Arg在敗血症後補充效果並不顯著，由於只在敗血症前補充Arg與敗血症前後均補充Arg的組別在各項分析結果均無統計上之差異，顯示在敗血症前後均補充Arg並無加成之效果。

關鍵字

敗血症；巨噬細胞；免疫球蛋白； Peyer’s patches ；精胺酸

並列摘要

This study investigated the effect of arginine (Arg) supplementation before or/and after sepsis on nutrient metabolism and mucosal immunity in rats. Male Wistar rats were assigned to four groups. Group 1 and 2 fed with semipurified diet, group 3 and 4 replaced part of casein with 2% of total calorie as Arg. After feeding the experimental diets for 10 days, sepsis was induced by cecal ligation and puncture (CLP), at the same time internal jugular vein was cannulated. All rats were maintained with total parenteral nutrition (TPN) for 3 days. Group 1 and 2 infused conventional TPN, while group 3 and 4 was supplemented with Arg, replacing 10% of total amino acid in TPN solution. The 4 groups were: group 1: without Arg supplementation before and after CLP, group 2: Arg supplemented after CLP, group 3: Arg supplemented before CLP, group 4: Arg supplemented before and after CLP. The experimental diets and TPN solutions were isonitogenous among the groups. All rats were sacrificed 3 days after CLP. The results showed that plasma glucose did not differ among groups, however, plasma triglyceride and free fatty acids were significantly lower in the group 3 and 4 than the other groups. Arg supplementation effectively increased plasma Arg levels, whereas there were no differences in branch chain amino acid and glutamine concentrations among the groups. These results suggest that Arg supplementation before CLP might have effect on reducing lipolysis induced by sepsis, but no beneficial effect was observed on attenuating muscle protein breakdown. Aerobic and anaerobic bacteria colony counts were significantly reduced and phagocytic activity of macrophage in peritoneal lavage fluid was enhanced in group 3 and 4 than the other 2 groups. Total number of Peyer’s patches on the intestine, lymphocyte counts in the Peyer’s patches, and total immunoglobulin A in intestinal lavage fluid were significantly higher in the group 4 than group 1 and 2. Also, interleukin-10 concentrations in mitogen-stimulated splenocytes were higher in the group 4 than the group 1 and 2. These results suggest that Arg supplementation before sepsis may enhance humoral and nonspecific immune response at the site of injury. Since there were no differences in various analytic measurements between group 3 and group 4, it seemed that synergistic effect was not obvious with Arg supplementation before and after CLP.

並列關鍵字

sepsis ； macrophage ； immunoglobulin A ； Peyer’s patches ； arginine

參考文獻

Akaki T, Sato K, Shimizu T, Sano C, Kajitani H, Dekio S and Tomioka H (1997) Effector molecules in expression of the antimicrobial activity of macrophages against Mycobacterium avium complex: roles of reactive nitrogen intermediates, reactive oxygen intermediates, and free fatty acids. J Leukocyte Biol 62:795-804

Barbul A.(1986) Arginine: biochemistry, physiology and therapeutic implications. J Parenter Eneter Nutr 10:227-238

Barbul A, Sisto DA, Wasserkrug HL and Efron G.(1981) Arginine stimulates lymphocyte immune response in healthy human beings. Surgery 90: 244-251

Barbul A, Wasserkrug HL, Seifter E, Rettura G, Levencon SM and Feron G.(1980)Immunostimulatory effects of arginine in normal and injured rats. J Surg Res 29:228-232

Barbul A, Wasserkrug HL, Yoshimura NN, Tao R and Efron G.(1984) High arginine levels in intravenous hyperalimentation abrogate post-traumatic immune suppression. J Surg Res 36:620-624

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添加精胺酸對腹膜炎引致敗血症老鼠腸道及全身性免疫反應之影響

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