东南大学学报（医学版）
東南大學學報（醫學版）
동남대학학보（의학판）
JOURNAL OF SOUTHEAST UNIVERSITY(MEDICAL SCIENCE EDITION)
2015年
1期
17-21
,共5页
戚勋%苑永辉%钟红珊%多久和阳%徐克
慼勛%苑永輝%鐘紅珊%多久和暘%徐剋
척훈%원영휘%종홍산%다구화양%서극
1-磷酸鞘氨醇%受体%基因表达%血管新生%血流恢复%后肢缺血%小鼠
1-燐痠鞘氨醇%受體%基因錶達%血管新生%血流恢複%後肢缺血%小鼠
1-린산초안순%수체%기인표체%혈관신생%혈류회복%후지결혈%소서
sphingosine 1-phosphate%receptor%gene expression%angiogenesis%blood flow recovery%ischemic hindlimb%mice
目的：：观察不同时间点小鼠后肢缺血模型肌肉组织中S1 P受体的基因表达情况,探讨其在后肢缺血模型血管新生及血流恢复中的作用。方法：将18只8~12周龄C57 Bl/6 J小鼠随机分为3 d组,即术后3 d组、术后7 d组及术后14 d组,每组6只。采用戊巴比妥麻醉小鼠,取仰卧位,钝性分离皮肤肌肉等外周组织后,分别结扎小鼠左后肢股动静脉及其分支后缝合,建立小鼠后肢缺血模型。分别于术后第1、3、5、7、10和14天采用激光多普勒血流分析仪观察缺血后肢血流变化情况。于术后第3、7和14天处死动物取腓肠肌组织行CD31免疫组织化学染色,计数微脉管密度,并提取腓肠肌组织中RNA,应用实时定量荧光PCR( real-time PCR)技术测定腓肠肌组织中S1P受体的mRNA表达情况。结果：术后14 d内,缺血侧与未缺血侧血流比值及缺血侧CD31阳性染色微脉管密度随时间改变逐渐增加。术后第3天,S1P1、S1P2和S1P3受体mRNA表达水平显著性增高,差异有统计学意义(P<0.05)；术后第7天和第14天,S1P2受体mRNA表达水平均显著地增高,差异具有统计学意义(P<0.05),而S1P1和S1P3受体mRNA表达水平无明显改变(P>0.05)。结论：小鼠后肢缺血后的血管新生和血流恢复情况在早期可能与S1P1受体、S1P2受体和S1P3受体上调有关,随后与S1P2受体上调有关。
目的：：觀察不同時間點小鼠後肢缺血模型肌肉組織中S1 P受體的基因錶達情況,探討其在後肢缺血模型血管新生及血流恢複中的作用。方法：將18隻8~12週齡C57 Bl/6 J小鼠隨機分為3 d組,即術後3 d組、術後7 d組及術後14 d組,每組6隻。採用戊巴比妥痳醉小鼠,取仰臥位,鈍性分離皮膚肌肉等外週組織後,分彆結扎小鼠左後肢股動靜脈及其分支後縫閤,建立小鼠後肢缺血模型。分彆于術後第1、3、5、7、10和14天採用激光多普勒血流分析儀觀察缺血後肢血流變化情況。于術後第3、7和14天處死動物取腓腸肌組織行CD31免疫組織化學染色,計數微脈管密度,併提取腓腸肌組織中RNA,應用實時定量熒光PCR( real-time PCR)技術測定腓腸肌組織中S1P受體的mRNA錶達情況。結果：術後14 d內,缺血側與未缺血側血流比值及缺血側CD31暘性染色微脈管密度隨時間改變逐漸增加。術後第3天,S1P1、S1P2和S1P3受體mRNA錶達水平顯著性增高,差異有統計學意義(P<0.05)；術後第7天和第14天,S1P2受體mRNA錶達水平均顯著地增高,差異具有統計學意義(P<0.05),而S1P1和S1P3受體mRNA錶達水平無明顯改變(P>0.05)。結論：小鼠後肢缺血後的血管新生和血流恢複情況在早期可能與S1P1受體、S1P2受體和S1P3受體上調有關,隨後與S1P2受體上調有關。
목적：：관찰불동시간점소서후지결혈모형기육조직중S1 P수체적기인표체정황,탐토기재후지결혈모형혈관신생급혈류회복중적작용。방법：장18지8~12주령C57 Bl/6 J소서수궤분위3 d조,즉술후3 d조、술후7 d조급술후14 d조,매조6지。채용무파비타마취소서,취앙와위,둔성분리피부기육등외주조직후,분별결찰소서좌후지고동정맥급기분지후봉합,건립소서후지결혈모형。분별우술후제1、3、5、7、10화14천채용격광다보륵혈류분석의관찰결혈후지혈류변화정황。우술후제3、7화14천처사동물취비장기조직행CD31면역조직화학염색,계수미맥관밀도,병제취비장기조직중RNA,응용실시정량형광PCR( real-time PCR)기술측정비장기조직중S1P수체적mRNA표체정황。결과：술후14 d내,결혈측여미결혈측혈류비치급결혈측CD31양성염색미맥관밀도수시간개변축점증가。술후제3천,S1P1、S1P2화S1P3수체mRNA표체수평현저성증고,차이유통계학의의(P<0.05)；술후제7천화제14천,S1P2수체mRNA표체수평균현저지증고,차이구유통계학의의(P<0.05),이S1P1화S1P3수체mRNA표체수평무명현개변(P>0.05)。결론：소서후지결혈후적혈관신생화혈류회복정황재조기가능여S1P1수체、S1P2수체화S1P3수체상조유관,수후여S1P2수체상조유관。
Objective:To investigate the expression of S1P receptor mRNA level in ischemic hindlimbs of mice at different time points, in order to discuss possible effects on angiogenesis and blood flow recovery. Methods:8-12 weeks old male C57Bl/6J mice were randomized divided into 3 groups:post-operative day 3 group, post-operative day 7 group and post-operative day 14 group, 6 mice for each group. Mice were subjected to surgical procedures to achieve unilateral hindlimb ischaemia after intraperitoneal injection of pentobarbital. In brief, following a skin incision at the left paracenter of the lower abdomen, the femoral artery, which originated from the external iliac artery and terminated to bifurcate into the saphenous and the popliteal arteries, was excised. The blood flow was observed at post-operative day 1, 3, 5, 7, 10 and 14 by laser Doppler blood flow analyzer. Ischemic tissue of calf muscle was excised at different time points for immunohistochemistry and real-time PCR to detect the capillary density and the expression of S1P receptor mRNA level. Results:The blood flow and capillary density in ischemic hindlimb were increased during the 14 days after operation. The mRNA expression of S1P1 and S1P3 was increased in ischemic limb muscle compared with non-ischemic limb muscle on post-operative day 3 ( P<0 . 05 ) , and there was no significant difference of S1P1 and S1P3 between ischemic hind limbs and non-ischemic hindlimbs on the post-operative day 7 and post-operative day 14(P>0. 05). There was significant difference of S1P2 mRNA level between ischemic hind limbs and non-ischemic hindlimbs on the post-operative day 3 , 7 and 14 ( P <0 . 05 ) . Conclusion:The mRNA expressions of S1P1 , S1P2 and S1P3 increase in the early time phase, and only S1P2 later in ischemic hindlimbs for angiogenesis and blood flow recovery.
