CAJ | 학술논문

背景:鱼油中的大量n-3多不饱和脂肪酸能抑制心脏移植物血管病变,延长移植物存活时间,但机制尚不完全清楚.新近研究提示,在体外实验中n-3多不饱和脂肪酸可通过活化过氧化物酶增殖子活化受体γ(peroxisome proliferator-activated receptor-γ,PPARγ)抑制炎性递质释放.目的:拟证实活化PPARγ否为n-3多不饱和脂肪酸缓解心脏移植物血管病变的新机制之一.方法:随机选取6只Lewis大鼠及18只Fisher344作为心脏供体.抽签法将另外24只Lewis大鼠随机分为4组作为受体,每组6只:同系移植组:Lewis大鼠之间心脏移植,不给予任何药物:低剂量鱼油组:行F344→Lewis异系移植,术后第1天开始按0.03 mL/kg给予鱼油灌胃8周;高剂量鱼油组:行F344→Lewis异系移植,术后第1天开始按0.06 mL/kg给予鱼油灌胃8周:对照组:行F344→Lewis异系移植,给予等体积环孢素A灌胃8周.低、高剂量鱼油组均在移植后开始环孢素A1.5mg/(kg·d)肌注2周.组织学检查评价心脏移植物血管病变,组织匀浆测定核因子kB及PPARγ活性,ELISA测定单核趋化蛋白1、干扰素诱导蛋白10含量,实时定量PT-PCR检测趋化因子受体CCR2及CXCR3表达.结果与结论:所有24只受体Lewis大鼠术后均顺利存活,供心在移植后8周仍有规律搏动.与同系移植组相比,移植8周后对照组心脏移植物发生了严重心脏移植物血管病变.与对照组相比,高、低剂量鱼油组心脏移植物血管病变均显著缓解,PPARγ性显著提高,核因子KB活性显著下降,组织匀浆单核趋化蛋白1及干扰素诱导蛋白10含量显著降低,趋化因子受体CCR2表达亦显著下调(P<0.001,P<0.05),高剂量鱼油组作用更明显(P<0.05).高、低剂量鱼油组及对照组CXCR3表达差异无显著性意义.实验证实,n-3多不饱和脂肪酸可通过活化核蛋白PPARγ制核因子kB活性与趋化因子及其受体表达,呈剂量依赖性缓解近交系大鼠模型心脏移植物血管病变的发生及发展. 揹景:魚油中的大量n-3多不飽和脂肪痠能抑製心髒移植物血管病變,延長移植物存活時間,但機製尚不完全清楚.新近研究提示,在體外實驗中n-3多不飽和脂肪痠可通過活化過氧化物酶增殖子活化受體γ(peroxisome proliferator-activated receptor-γ,PPARγ)抑製炎性遞質釋放.目的:擬證實活化PPARγ否為n-3多不飽和脂肪痠緩解心髒移植物血管病變的新機製之一.方法:隨機選取6隻Lewis大鼠及18隻Fisher344作為心髒供體.抽籤法將另外24隻Lewis大鼠隨機分為4組作為受體,每組6隻:同繫移植組:Lewis大鼠之間心髒移植,不給予任何藥物:低劑量魚油組:行F344→Lewis異繫移植,術後第1天開始按0.03 mL/kg給予魚油灌胃8週;高劑量魚油組:行F344→Lewis異繫移植,術後第1天開始按0.06 mL/kg給予魚油灌胃8週:對照組:行F344→Lewis異繫移植,給予等體積環孢素A灌胃8週.低、高劑量魚油組均在移植後開始環孢素A1.5mg/(kg·d)肌註2週.組織學檢查評價心髒移植物血管病變,組織勻漿測定覈因子kB及PPARγ活性,ELISA測定單覈趨化蛋白1、榦擾素誘導蛋白10含量,實時定量PT-PCR檢測趨化因子受體CCR2及CXCR3錶達.結果與結論:所有24隻受體Lewis大鼠術後均順利存活,供心在移植後8週仍有規律搏動.與同繫移植組相比,移植8週後對照組心髒移植物髮生瞭嚴重心髒移植物血管病變.與對照組相比,高、低劑量魚油組心髒移植物血管病變均顯著緩解,PPARγ性顯著提高,覈因子KB活性顯著下降,組織勻漿單覈趨化蛋白1及榦擾素誘導蛋白10含量顯著降低,趨化因子受體CCR2錶達亦顯著下調(P<0.001,P<0.05),高劑量魚油組作用更明顯(P<0.05).高、低劑量魚油組及對照組CXCR3錶達差異無顯著性意義.實驗證實,n-3多不飽和脂肪痠可通過活化覈蛋白PPARγ製覈因子kB活性與趨化因子及其受體錶達,呈劑量依賴性緩解近交繫大鼠模型心髒移植物血管病變的髮生及髮展.
배경:어유중적대량n-3다불포화지방산능억제심장이식물혈관병변,연장이식물존활시간,단궤제상불완전청초.신근연구제시,재체외실험중n-3다불포화지방산가통과활화과양화물매증식자활화수체γ(peroxisome proliferator-activated receptor-γ,PPARγ)억제염성체질석방.목적:의증실활화PPARγ부위n-3다불포화지방산완해심장이식물혈관병변적신궤제지일.방법:수궤선취6지Lewis대서급18지Fisher344작위심장공체.추첨법장령외24지Lewis대서수궤분위4조작위수체,매조6지:동계이식조:Lewis대서지간심장이식,불급여임하약물:저제량어유조:행F344→Lewis이계이식,술후제1천개시안0.03 mL/kg급여어유관위8주;고제량어유조:행F344→Lewis이계이식,술후제1천개시안0.06 mL/kg급여어유관위8주:대조조:행F344→Lewis이계이식,급여등체적배포소A관위8주.저、고제량어유조균재이식후개시배포소A1.5mg/(kg·d)기주2주.조직학검사평개심장이식물혈관병변,조직균장측정핵인자kB급PPARγ활성,ELISA측정단핵추화단백1、간우소유도단백10함량,실시정량PT-PCR검측추화인자수체CCR2급CXCR3표체.결과여결론:소유24지수체Lewis대서술후균순리존활,공심재이식후8주잉유규률박동.여동계이식조상비,이식8주후대조조심장이식물발생료엄중심장이식물혈관병변.여대조조상비,고、저제량어유조심장이식물혈관병변균현저완해,PPARγ성현저제고,핵인자KB활성현저하강,조직균장단핵추화단백1급간우소유도단백10함량현저강저,추화인자수체CCR2표체역현저하조(P<0.001,P<0.05),고제량어유조작용경명현(P<0.05).고、저제량어유조급대조조CXCR3표체차이무현저성의의.실험증실,n-3다불포화지방산가통과활화핵단백PPARγ제핵인자kB활성여추화인자급기수체표체,정제량의뢰성완해근교계대서모형심장이식물혈관병변적발생급발전.
BACKGROUND: Fish oil is one of mainly natural resources of n-3 fatty acids, which can inhibit cardiac allograft vasculopathy (CAV) and prolong the survival of cardiac allograft. But, the mechanism is unclear. Recent in vitro data suggested that n-3 fatty acids could inhibit the release of inflammatory transmitter by the activation of peroxisome proliferator-activated receptor-y (PPARy). OBJECTIVE: To test the hypothesis that n-3 fatty acids from fish oil ameliorates CAV development via activating PPARy. METHODS: A total of 6 Lewis rats and 18 Fisher344 rats were randomly selected as heart donors. An additional 24 Lewis rats were randomly and equally divided into 4 groups. In isograft group, heart transplantation was performed among Lewis rats, without any drug. In low-dose fish oil-treated group, F344→Lewis transplantation was performed. At 1 day following surgery, 0.03 mL/kg fish oil was treated by gavage for 8 weeks. In high-dose fish oil-treated group, F344→Lewis transplantation was conducted. At 1 day following surgery, 0.06 mL/kg fish oil was treated by gavage for 8 weeks. In control group, F344→Lawis transplantation was conducted. Cyclosporine A was administrated by gavage for 8 weeks. In the low-dose and high-dose fish oil-treated groups, cyclosporine A (1.5 mg/kg) was given daily by intramuscular injection for 2 weeks following surgery. CAV was evaluated by histological examination. Activity of nuclear factor (NF) k-B and PPARy was assessed in homogenate. Contents of monocyte chemoattractant protein-1 and interferon-inducible protein 10 were measured by enzyme-labeled immunosorbent assay (ELISA). Chemokine receptor CCR2 and CXCR3 expression was determined by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR). RESULTS AND CONCLUSION: All 24 receptor Lewis rats were survived following surgery. The donor heart could regularly beat at 8 weeks following transplantation. Compared with the isograft group, severe CAV was detected in the control group al 8 weeks. Compared with the control group, CAV was significantly relieved, the activity of PPARy was significantly elevated, the activity of NF k-B was significantly decreased, levels of intragraft monocyte chemoattractant protein-1 and interferon-inducible protein-10 were significantly reduced in the low-dose and high-dose fish oil-treated groups (P < 0.001, P < 0.05), especially in the high-dose fish oil-treated group (P < 0.05). There was no significant difference in expression of chemokine receptors CXCR3 in the low-dose and high-dose fish oil-treated groups and control group. Our results demonstrated that n-3 fatty acids from fish oil can attenuate CAV development, possibly through activating PPARy and subsequently inhibiting the NF-kB activation, the chemokines secretion and its receptor expression in a dose-dependent fashion in rat models.