中华器官移植杂志
中華器官移植雜誌
중화기관이식잡지
CHINESE JOURNAL OF ORGAN TRANSPLANTATION
2010年
3期
157-161
,共5页
张松林%孙宗全%冯剑锷%吴龙%于利
張鬆林%孫宗全%馮劍鍔%吳龍%于利
장송림%손종전%풍검악%오룡%우리
一氧化碳%核因子出%κB%炎症%凋亡%缺血再灌注损伤%心脏移植
一氧化碳%覈因子齣%κB%炎癥%凋亡%缺血再灌註損傷%心髒移植
일양화탄%핵인자출%κB%염증%조망%결혈재관주손상%심장이식
Carbon monoxide%Nuclear factor kappa B%Inflammation%Apoptosis%Ischemia/reperfusion injury%Heart transplantation
目的 观察心脏移植前用二氯甲烷(MC)灌胃增加受者体内一氧化碳含量在减轻小鼠移植心缺血再灌注损伤中的作用及其机制.方法 以Balb/c小鼠作为供、受者,建立颈部异位心脏移植模型.实验共分为4组,分别为MC 100 mg组(n=10)、MC 500 mg组(n=12)、橄榄油组(n=10)和正常对照组(n=5).前3组在麻醉前3 h分别用经橄榄油稀释的MC 100 nag/kg、MC 500mg/kg及单纯橄榄油0.15 ml对受者进行灌胃处理,然后行颈部异位心脏移植;正常对照组小鼠仅作麻醉处理,不进行心脏移植.分别于灌胃后0、1、3、6、12和24 h剪尾采血,测定血液中CO的含量[用碳氧血红蛋白(COHb)占总血红蛋白的百分比表示],并于相应时间点取心肌组织,检测心肌组织中CO的含量;各组移植后3和24 h分别处死半数受者,检测移植后3和24 h血清中肌钙蛋白I(cT-nI)、心肌组织中肿瘤坏死因子α(TNF-α)、白细胞介素10(IL-10)、细胞凋亡与相关基因Bcl-2和Bax mRNA以及核因子κB(NF-κB)的表达,并观察移植心心肌的超微结构.结果 与橄榄油组比较,MC 100 mg和MC 500 mg组受者血液中COHb浓度与心肌组织中CO含量明显升高,均在灌胃后3h达到峰值;MC 100 mg组和MC 500mg组受者心脏移植后3和24h,能显著降低血清中cTnI水平(P<0.01),并以MC 500 mg组降低最为明显;可明显下调促炎症基因TNF-α mRNA水平(P<0.01),而抗炎症基因IL-10 mRNA上调不明显(P>0.05),同时可以显著上调抗凋亡基因Bcl-2 mRNA水平(P<0.01),并抑制促凋亡基因Bax mRNA的转录(P<0.01);心肌超微结构损伤明显减轻.正常对照组可见少量NF-κB表达,而橄榄油组、MC 100 mg组和MC 500 mg组NF-κB活性均显著增强(P<0.01),但后三组之间NF-κB活性的差异无统计学意义(P>0.05).结论 诱导受者体内增加CO含量能通过其抗炎症和抗凋亡功能而减轻移植心缺血再灌注损伤;但与抑制NF-κB信号转导通路无关.
目的 觀察心髒移植前用二氯甲烷(MC)灌胃增加受者體內一氧化碳含量在減輕小鼠移植心缺血再灌註損傷中的作用及其機製.方法 以Balb/c小鼠作為供、受者,建立頸部異位心髒移植模型.實驗共分為4組,分彆為MC 100 mg組(n=10)、MC 500 mg組(n=12)、橄欖油組(n=10)和正常對照組(n=5).前3組在痳醉前3 h分彆用經橄欖油稀釋的MC 100 nag/kg、MC 500mg/kg及單純橄欖油0.15 ml對受者進行灌胃處理,然後行頸部異位心髒移植;正常對照組小鼠僅作痳醉處理,不進行心髒移植.分彆于灌胃後0、1、3、6、12和24 h剪尾採血,測定血液中CO的含量[用碳氧血紅蛋白(COHb)佔總血紅蛋白的百分比錶示],併于相應時間點取心肌組織,檢測心肌組織中CO的含量;各組移植後3和24 h分彆處死半數受者,檢測移植後3和24 h血清中肌鈣蛋白I(cT-nI)、心肌組織中腫瘤壞死因子α(TNF-α)、白細胞介素10(IL-10)、細胞凋亡與相關基因Bcl-2和Bax mRNA以及覈因子κB(NF-κB)的錶達,併觀察移植心心肌的超微結構.結果 與橄欖油組比較,MC 100 mg和MC 500 mg組受者血液中COHb濃度與心肌組織中CO含量明顯升高,均在灌胃後3h達到峰值;MC 100 mg組和MC 500mg組受者心髒移植後3和24h,能顯著降低血清中cTnI水平(P<0.01),併以MC 500 mg組降低最為明顯;可明顯下調促炎癥基因TNF-α mRNA水平(P<0.01),而抗炎癥基因IL-10 mRNA上調不明顯(P>0.05),同時可以顯著上調抗凋亡基因Bcl-2 mRNA水平(P<0.01),併抑製促凋亡基因Bax mRNA的轉錄(P<0.01);心肌超微結構損傷明顯減輕.正常對照組可見少量NF-κB錶達,而橄欖油組、MC 100 mg組和MC 500 mg組NF-κB活性均顯著增彊(P<0.01),但後三組之間NF-κB活性的差異無統計學意義(P>0.05).結論 誘導受者體內增加CO含量能通過其抗炎癥和抗凋亡功能而減輕移植心缺血再灌註損傷;但與抑製NF-κB信號轉導通路無關.
목적 관찰심장이식전용이록갑완(MC)관위증가수자체내일양화탄함량재감경소서이식심결혈재관주손상중적작용급기궤제.방법 이Balb/c소서작위공、수자,건립경부이위심장이식모형.실험공분위4조,분별위MC 100 mg조(n=10)、MC 500 mg조(n=12)、감람유조(n=10)화정상대조조(n=5).전3조재마취전3 h분별용경감람유희석적MC 100 nag/kg、MC 500mg/kg급단순감람유0.15 ml대수자진행관위처리,연후행경부이위심장이식;정상대조조소서부작마취처리,불진행심장이식.분별우관위후0、1、3、6、12화24 h전미채혈,측정혈액중CO적함량[용탄양혈홍단백(COHb)점총혈홍단백적백분비표시],병우상응시간점취심기조직,검측심기조직중CO적함량;각조이식후3화24 h분별처사반수수자,검측이식후3화24 h혈청중기개단백I(cT-nI)、심기조직중종류배사인자α(TNF-α)、백세포개소10(IL-10)、세포조망여상관기인Bcl-2화Bax mRNA이급핵인자κB(NF-κB)적표체,병관찰이식심심기적초미결구.결과 여감람유조비교,MC 100 mg화MC 500 mg조수자혈액중COHb농도여심기조직중CO함량명현승고,균재관위후3h체도봉치;MC 100 mg조화MC 500mg조수자심장이식후3화24h,능현저강저혈청중cTnI수평(P<0.01),병이MC 500 mg조강저최위명현;가명현하조촉염증기인TNF-α mRNA수평(P<0.01),이항염증기인IL-10 mRNA상조불명현(P>0.05),동시가이현저상조항조망기인Bcl-2 mRNA수평(P<0.01),병억제촉조망기인Bax mRNA적전록(P<0.01);심기초미결구손상명현감경.정상대조조가견소량NF-κB표체,이감람유조、MC 100 mg조화MC 500 mg조NF-κB활성균현저증강(P<0.01),단후삼조지간NF-κB활성적차이무통계학의의(P>0.05).결론 유도수자체내증가CO함량능통과기항염증화항조망공능이감경이식심결혈재관주손상;단여억제NF-κB신호전도통로무관.
Objective To examine whether the increase of carbon monoxide (CO) induced by oral methylene chloride (MC) administration in recipients before heart transplantation would protect heart grafts against isehemia/reperfusion (I/R) injury associated with transplantation and to explore the possible mechanism.Methods Inbred male Balb/c mice were used as donors and recipients to establish cervical heart transplantation model Recipients were treated with either MC (100 mg/kg or 500 mg/kg,per os)(group MC 100 mg,n=10;group MC 500 mg,n=12) or olive oil(0.15 ml,per os.group olive,n=10) 3 h prior to anesthesia.Age-matched norwlal mice served as controls (group N,n=5).The serum COHb and the CO content of myocardial tissue were measured at 0,1,3,6,12,24 h after oral MC administration.Half of recipients were killed at 3 and 24h after transplantation for senum or cardiac graft samples.The serum cTnI levels,the mRNA levels of TNF-α,IL-10,Bcl-2,Bax.the protein levels of NF-κB and the ultrastructures of myocardium were examined.Results As tompared with group olive.the serum COHb and tissue CO were increased significantly and peaked within 3 h in group MC 100 mg and group MC 500 mg.The serum cTnI levels in group MC 100 mg and group MC 500 mg were significantly decreased (P<0. 01 ), especially in group MC 500 mg. The increase of CO in recipients of group MC100 mg and group MC 500 mg significantly inhibited the proinflammatory gene expression of TNF-α mRNA and the pro-apoptotic gene expression of Bax mRNA (P<0. 01), and increased the anti-apoptotic gene expression of Bcl-2 mRNA (P<0. 01), but did not increase the anti-inflammatory gene expression of IL-10 mRNA (P>0. 05) in the heart grafts. As compared with group N, the myocardial NF-κB activation was increased significantly in group olive,group MC 100 mg and group MC 500 mg (P<0. 01 ), but there was no significant difference among the later three groups (P>0. 05). The myocardial ultrastructure was also alleviated significantly in group MC 100 mg and group MC 500 mg as compared with group N. Conclusion The increase of CO induced by MC in recipients suppresses pro-inflammatory and pro-apoptotic gene expression and efficiently ameliorates transplant-induced heart I/R injury. The possible mechanism does not seem to be associated with down-regulation of the NF-κB signaling pathway.
