不同缺血預(yù)處理方案對(duì)大鼠肢體缺血再灌注損傷保護(hù)效應(yīng)的實(shí)驗(yàn)研究
發(fā)布時(shí)間：2018-07-20 14:14
【摘要】：目的缺血再灌注損傷在臨床較為常見(jiàn)，防治措施眾多，但多數(shù)效果不確切，本實(shí)驗(yàn)旨在探討缺血預(yù)處理不同時(shí)間方案對(duì)肢體缺血再灌注損傷保護(hù)作用的差異性，選擇合理的缺血預(yù)處理時(shí)間，為臨床開(kāi)展缺血預(yù)處理技術(shù)提供充足的理論依據(jù)。 方法取40只健康SPF級(jí)SD大鼠，隨機(jī)分成5組（n=8）。術(shù)前禁食12h，禁水2h。腹腔10%水合氯醛麻醉后分離顯露腹主動(dòng)脈下段和下腔靜脈，在腸系膜下動(dòng)脈與雙髂動(dòng)脈分叉間用微血管夾阻斷腹主動(dòng)脈血流2h、復(fù)通血流2h為缺血再灌注損傷模型。假手術(shù)組（A組，僅行開(kāi)腹，分離腹主動(dòng)脈不阻斷血流）；缺血再灌注組（B組，，夾閉腹主動(dòng)脈缺血2h后再灌注2h）；C、D、E組：分別阻斷腹主動(dòng)脈1、5和10min，再灌注1、5和10min，如此重復(fù)3個(gè)循環(huán)后再進(jìn)行2h缺血2h再灌注。各組均在實(shí)驗(yàn)程序結(jié)束復(fù)通血流后經(jīng)下腔靜脈采血34ml，3000r/min離心15min后-80℃保存。采用硫代巴比妥酸(TBA)法測(cè)定MDA濃度，黃嘌呤氧化酶法測(cè)定SOD活性，使用雙抗體一步夾心酶聯(lián)免疫吸附試驗(yàn)法（ELISA）測(cè)定血清超氧化物歧化酶（SOD）、丙二醛（MDA）、白細(xì)胞介素-6（IL-6）、腫瘤壞死因子-α（TNF-α）、白細(xì)胞介素-10（IL-10）水平，觀察各組氧化/抗氧化指標(biāo)及炎癥因子表達(dá)的差異性。 實(shí)驗(yàn)結(jié)果采用SPSS19.0統(tǒng)計(jì)軟件進(jìn)行分析，計(jì)量資料以x_±s表示，所有分組均進(jìn)行K-S檢驗(yàn)、方差齊性檢驗(yàn)，組間兩兩比較采用LSD法單因素方差分析。 結(jié)果 1.與A組比較，B組的SOD活力下降，MDA含量升高，差異有統(tǒng)計(jì)學(xué)意義（P＜0.05）；與B組比較，C、D、E組的SOD活力升高，MDA含量下降，其中C組與B組比較的差異無(wú)統(tǒng)計(jì)學(xué)意義（P＞0.05），D、E組與B組比較差異有統(tǒng)計(jì)學(xué)意義（P＜0.05）；C、D、E進(jìn)行組間比較，E組的SOD活力最高、MDA含量最低，C組SOD活力最低、MDA含量最高，各組間差異有統(tǒng)計(jì)學(xué)意義（P＜0.05）。 2.與A組比較，B組的IL-6、TNF-α、IL-10含量升高，差異均有統(tǒng)計(jì)學(xué)意義（P＜0.05）；與B組比較，C、D、E組的IL-6、TNF-α含量明顯升高，差異均有統(tǒng)計(jì)學(xué)意義（P＜0.05）；C、D、E進(jìn)行組間比較，各數(shù)值均呈遞增趨勢(shì)，IL-6的含量D組和C組比較、E組和D組比較差異均無(wú)統(tǒng)計(jì)學(xué)意義（P＞0.05）。TNF-α含量D組和C組差異無(wú)統(tǒng)計(jì)學(xué)意義（P＞0.05），E組和D組差異有統(tǒng)計(jì)學(xué)意義（P＜0.05）。IL-10含量各組間進(jìn)行比較差異均有統(tǒng)計(jì)學(xué)意義（P＜0.05）。 結(jié)論 1.缺血預(yù)處理對(duì)缺血再灌注誘發(fā)的氧化損傷存在保護(hù)作用，在IprC1、5、10min/3個(gè)循環(huán)分組內(nèi)顯示IprC時(shí)間與其保護(hù)作用呈平行關(guān)系。 2.機(jī)體在缺血預(yù)處理時(shí)會(huì)發(fā)生炎癥反應(yīng)，IprC時(shí)間的增加促進(jìn)了炎癥因子的表達(dá)和蓄積，顯示IprC在肢體缺血所致的炎癥反應(yīng)上無(wú)明顯保護(hù)作用，甚至可增加肢體缺血最終造成全身性炎癥反應(yīng)。 3.本實(shí)驗(yàn)顯示，1min/3個(gè)循環(huán)缺血預(yù)處理方案發(fā)揮的抗氧化損傷作用無(wú)統(tǒng)計(jì)學(xué)意義，誘導(dǎo)的炎癥反應(yīng)輕微。10min/3個(gè)循環(huán)方案抗氧化損傷作用最強(qiáng)，但是炎癥反應(yīng)最重，不適宜選用。5min/3個(gè)循環(huán)方案有明顯的抗氧化損傷作用，炎癥反應(yīng)適中，綜合來(lái)看臨床選作缺血預(yù)處理策略較為適宜。
[Abstract]:Objective ischemia reperfusion injury is more common in clinical practice, and the prevention and treatment measures are numerous, but most of the effects are not accurate. The purpose of this experiment is to explore the difference in the protective effect of ischemic preconditioning on the protection of ischemia reperfusion injury of limb, and choose the reasonable time of ischemic preconditioning to provide sufficient theoretical basis for the development of ischemic preconditioning in bed. According to it.
Methods 40 healthy SPF grade SD rats were randomly divided into 5 groups (n=8). Before operation, 12h was fasted and the lower abdominal aorta and inferior vena cava were exposed after 2h. abdominal 10% hydrated chloral anaesthesia. The blood flow 2H was blocked by microvascular clamp between the inferior mesenteric artery and the double iliac artery, and the revascularised 2H was the model of ischemia reperfusion injury. The sham operation was performed. Group A (group B, isolated abdominal aorta without blocking blood flow), ischemia reperfusion group (group B, occlusion of abdominal aorta ischemia 2H reperfusion 2H); C, D, E group: block the abdominal aorta 1,5 and 10min, reperfusion 1,5 and 10min, then repeat the 3 cycles and repeat the 2H ischemia-reperfusion. The blood sampling of the inferior vena cava was 34ml, 3000r/min was preserved at -80 C after 15min. The concentration of MDA was measured by thiobarbituric acid (TBA), the activity of SOD was measured by xanthine oxidase method, and the serum superoxide dismutase (SOD), malondialdehyde (MDA), -6 (IL-6), and the cause of the tumor's bad death were determined by the double antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sub - alpha (TNF- alpha) and interleukin -10 (IL-10) levels were observed to observe the difference of oxidation / antioxidant index and inflammatory factors expression in each group.
The experimental results were analyzed with SPSS19.0 statistical software, and the measurement data were expressed in x_ + s. All the groups were tested with K-S test, variance homogeneity test, and 22 of groups were compared by LSD single factor analysis of variance.
Result
1. compared with the A group, the activity of SOD in the B group decreased and the MDA content increased, and the difference was statistically significant (P < 0.05). Compared with the B group, the SOD activity of C, D and E decreased, and the difference between the C group and the group was not statistically significant (0.05). D activity was the highest, MDA content was the lowest, C group SOD activity was lowest, MDA content was the highest, the difference was statistically significant among all groups (P < 0.05).
2. compared with the A group, the content of IL-6, TNF- a, IL-10 in group B increased, the difference was statistically significant (P < 0.05). Compared with the B group, C, D, E group were significantly higher, the difference was statistically significant (0.05). There was no statistical significance (P > 0.05).TNF- alpha content in D group and C group (P > 0.05), and there was significant difference between group E and D group (P < 0.05), and there was significant difference between each group (P < 0.05).
conclusion
1. ischemic preconditioning has a protective effect on the oxidative damage induced by ischemia-reperfusion, and the IprC time in the IprC1,5,10min/3 cycle grouping is parallel to its protective effect.
The 2. body will have an inflammatory response during ischemic preconditioning. The increase of IprC time promotes the expression and accumulation of inflammatory factors. It shows that IprC has no obvious protective effect on the inflammatory response to limb ischemia, and may even increase limb ischemia and eventually cause systemic inflammatory response.
3. the experiment showed that the anti oxidative damage effect of 1min/3 cyclic ischemic preconditioning regimen was not statistically significant. The mild.10min/3 cycle regimen induced by the induced inflammatory reaction was the strongest, but the inflammatory reaction was the heaviest. The.5min/3 cycle scheme was not suitable for anti oxidative damage. The inflammatory reaction was moderate, and the inflammatory reaction was moderate. In conclusion, the ischemic preconditioning strategy is more suitable.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R54
本文編號(hào)：2133802
[Abstract]:Objective ischemia reperfusion injury is more common in clinical practice, and the prevention and treatment measures are numerous, but most of the effects are not accurate. The purpose of this experiment is to explore the difference in the protective effect of ischemic preconditioning on the protection of ischemia reperfusion injury of limb, and choose the reasonable time of ischemic preconditioning to provide sufficient theoretical basis for the development of ischemic preconditioning in bed. According to it.
Methods 40 healthy SPF grade SD rats were randomly divided into 5 groups (n=8). Before operation, 12h was fasted and the lower abdominal aorta and inferior vena cava were exposed after 2h. abdominal 10% hydrated chloral anaesthesia. The blood flow 2H was blocked by microvascular clamp between the inferior mesenteric artery and the double iliac artery, and the revascularised 2H was the model of ischemia reperfusion injury. The sham operation was performed. Group A (group B, isolated abdominal aorta without blocking blood flow), ischemia reperfusion group (group B, occlusion of abdominal aorta ischemia 2H reperfusion 2H); C, D, E group: block the abdominal aorta 1,5 and 10min, reperfusion 1,5 and 10min, then repeat the 3 cycles and repeat the 2H ischemia-reperfusion. The blood sampling of the inferior vena cava was 34ml, 3000r/min was preserved at -80 C after 15min. The concentration of MDA was measured by thiobarbituric acid (TBA), the activity of SOD was measured by xanthine oxidase method, and the serum superoxide dismutase (SOD), malondialdehyde (MDA), -6 (IL-6), and the cause of the tumor's bad death were determined by the double antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sub - alpha (TNF- alpha) and interleukin -10 (IL-10) levels were observed to observe the difference of oxidation / antioxidant index and inflammatory factors expression in each group.
The experimental results were analyzed with SPSS19.0 statistical software, and the measurement data were expressed in x_ + s. All the groups were tested with K-S test, variance homogeneity test, and 22 of groups were compared by LSD single factor analysis of variance.
Result
1. compared with the A group, the activity of SOD in the B group decreased and the MDA content increased, and the difference was statistically significant (P < 0.05). Compared with the B group, the SOD activity of C, D and E decreased, and the difference between the C group and the group was not statistically significant (0.05). D activity was the highest, MDA content was the lowest, C group SOD activity was lowest, MDA content was the highest, the difference was statistically significant among all groups (P < 0.05).
2. compared with the A group, the content of IL-6, TNF- a, IL-10 in group B increased, the difference was statistically significant (P < 0.05). Compared with the B group, C, D, E group were significantly higher, the difference was statistically significant (0.05). There was no statistical significance (P > 0.05).TNF- alpha content in D group and C group (P > 0.05), and there was significant difference between group E and D group (P < 0.05), and there was significant difference between each group (P < 0.05).
conclusion
1. ischemic preconditioning has a protective effect on the oxidative damage induced by ischemia-reperfusion, and the IprC time in the IprC1,5,10min/3 cycle grouping is parallel to its protective effect.
The 2. body will have an inflammatory response during ischemic preconditioning. The increase of IprC time promotes the expression and accumulation of inflammatory factors. It shows that IprC has no obvious protective effect on the inflammatory response to limb ischemia, and may even increase limb ischemia and eventually cause systemic inflammatory response.
3. the experiment showed that the anti oxidative damage effect of 1min/3 cyclic ischemic preconditioning regimen was not statistically significant. The mild.10min/3 cycle regimen induced by the induced inflammatory reaction was the strongest, but the inflammatory reaction was the heaviest. The.5min/3 cycle scheme was not suitable for anti oxidative damage. The inflammatory reaction was moderate, and the inflammatory reaction was moderate. In conclusion, the ischemic preconditioning strategy is more suitable.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R54
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