大蒜精油及其有機含硫組成分對於內毒素誘發大鼠腸道細菌移轉之影響及相關機制之探討(III)

中山醫學大學機構典藏 CSMUIR > 健康管理學院 > 營養學系暨碩士班 > 研究計劃 > Item 310902500/3214

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题名:	大蒜精油及其有機含硫組成分對於內毒素誘發大鼠腸道細菌移轉之影響及相關機制之探討(III) Studies of the Effects of Garlic Oil and Its Sulful-Containing Composition on Endotoxin-Induced Intestinal Bacteria Translocation and the Related Mechanisms (III)
作者:	劉承慈 Liu, Cheng-Tzu
贡献者:	中山醫學大學營養科學研究所
关键词:	大蒜精油;有機含硫成分;脂多糖;嗜中性球;自由基 Garlic oil;OSCs;Lipopolysaccharide;Neutrophil;Free radical
日期:	2003
上传时间:	2010-12-16T03:31:24Z (UTC)
摘要:	系統性發炎 (SIRS)時嗜中性球產生大量自由基，造成腸黏膜受損，增加腸道菌伺機感染的機會，已知是加重重症病患系統性發炎反應的重要原因之一，且此現象受到促發炎細胞激素之調節。本研究用雄性 Wistar 大鼠以載劑玉米油（CO， 2ml / kg BW）或大蒜精油（GO，50 及 200 mg / kg BW）及其有機含硫成分（0.5 mmol / kg BW DAS、DADS 或 DATS）灌食兩週後，由腹腔注射 5mg/kg 脂多糖（ LPS）誘發系統性發炎，同時禁食直到犧牲。在誘發後 2 或 3h 抽尾靜脈血，測定 TNF- α, IL-1β, IL-6 濃度, 且在 18h 後犧牲分析周邊血總白血球數及嗜中性球數，並分離嗜中性球，測量 PMA 刺激細胞釋放超氧自由基能力及趨化指數，另外也評估腸道組織嗜中性球浸潤及受損情形。此外, 亦觀察這些大蒜成分在活體外對於促發炎細胞激素刺激下的大鼠小腸上皮細胞 (IEC)存活率之影響. 結果顯示，未灌食大蒜下， LPS 刺激組相較於控制組之血中細胞激素濃度、總白血球數、嗜中性球數、自由基釋放能力、穿透能力、趨化指數、腸道嗜中性球浸潤及黏膜受損情形皆顯著上升（ p<0.05）。且在活體外促發炎細胞激素使 IEC 存活率下降（ p<0.05）。在 LPS 刺激條件下，DAS 可顯著抑制 IL-1 β濃度, 但是 200 mg/kg GO 及 DATS 顯著提高三種細胞激素濃度（p<0.05）。50mg/kg GO 顯著降低血中嗜中性球數、自由基釋放能力、腸道嗜中性球浸潤及腸道細胞 apoptosis （ p<0.05），而趨化指數及黏膜受損情形則有減緩的趨勢。DAS 及 DADS 不影響嗜中性球生理功能及腸道受損情形，但 DATS 顯著增加血中總白血球數、嗜中性球數、穿透能力、超氧自由基釋放能力、趨化指數、腸道嗜中性球浸潤及腸道細胞 apoptosis（p<0.05）。且活體外實驗亦發現 IEC 在促發炎性細胞激素存在下對於 DATS 細胞毒性的感受性增加。由本研究結果可知，在 LPS 誘發系統性發炎反應時，血中促發炎激素濃度、總白血球數及嗜中性球數、自由基釋放能力及趨化反應的上升導致腸道嗜中性球浸潤及黏膜受損。而低劑量 GO 可藉由降低嗜中性球數及其超氧自由基釋放能力而減緩發炎組織傷害程度，然而， DATS 則會增加血中促發炎激素濃度、嗜中性球數、嗜中性球超氧自由基釋放能力及其趨化反應，而使得腸道損傷加劇。 It is known that in SIRS, increased free radical from neutrophils regulated by proinflammatory cytokines raises the risk of opportunistic infection that deteriorates the clinical condition in such patients. The present study was carried out with male Wistar rats. All animals were were gavaged 50 or 200 mg/kg body wt garlic oil, or 0.5mmol/kg body weight OSCs of GO (DAS, DADS, or DATS), control animals were gavaged vehicle (corn oil, 2 mL/kg body wt) alone every other day for 2 weeks fallowed by the induction of SIRS by the injection of LPS (5mg/kg; i.p.). Food was removed at the time when the injection carried out until animals been killed.The control rat was injected with the same volume of vehicle. Blood was withdrawn from a lateral tail vein at 2 or 3 hours after the injection for the determination of TNF-.alpha., IL-1.beta., IL-6. All animals were killed at 18h after the injection and determined for the total white count and neutrophil count in peripheral blood. Nutrophils were isolated and determined for PMA-stimulated production of superoxide free radical and transmigration. Histological investigation was carried out with small intestine for neutrophil perfiltration and intestinal damage. The effects of these garlic compounds on the viability of IEC from rats in the absence or presence of inflammatory cytokines were also investigated in vitro. We found that LPS injection significantly elevated the concentration of inflammatory cytokines, total white count, neutrophil count, the capability of free radical production and transmigration in PMA-stimulated neutrophil, neutrophil perfiltration and apoptosis in intestine (P<0.05). In addition, the presence of proinflammatory cytokines decreases the viability of IEC in vitro (P<0.05). DAS significantly suppressed LPS-induced increase of IL-1 ？] level, however, 200 mg/kg GO and DATS significantly elevated the levels of all three cytokines in blood (P<0.05). GO fed at the dose of 50mg/kg significantly lowered neutrophil count and the capability of free radical production and transmigration of neutrophil, and significantly ameliorated neutrophil perfiltration and apoptosis in the intestine (P<0.05). Although DAS and DADS did not show any effect on the function of neutrophil and on intestine, DATS significantly elevated total white count and neutrophil count in blood and improved the capability of free radical production and transmigration of neutrophil, and significantly deteriorated neutrophil perfiltration and apoptosis in the intestine (P<0.05). In vitro study also found that the sensitivity of IEC to the cytotoxicity of DATS was increased in the presence of proinflammatory cytokines. In conclusion, LPS-injection elevated the concentration of serum proinflammatory cytokines, total white count and neutrophil count in blood and the capability of free radical production and transmigration of neutrophil in association with neutrophil perfiltration and apoptosis in intestinal mucosa. GO fed at the dose of 50mg/kg GO ameliorated LPS-induced neutrophil perfiltration and apoptosis in intestinal mucosa in associated with lowered neutrophil count and the capability of free radical production of neutrophil, however, DATS deteriorated LPS-induced neutrophil perfiltration and apoptosis in intestinal mucosa in association with elevated serum concentration of inflammatory cytokines, neutrophil count and the capability of free radical production and transmigration of neutrophil.
URI:	https://ir.csmu.edu.tw:8080/handle/310902500/3214
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