發(fā)布時間：2018-05-15 23:37

  本文選題：低溫保存 + 心肌細(xì)胞�。� 參考：《中國醫(yī)科大學(xué)》2007年博士論文

【摘要】： 低溫下心肌細(xì)胞骨架微管的變化與冷缺血損傷相關(guān)性的研究 目的 心臟移植是公認(rèn)的治療終末期心臟病唯一有效的手段,但由于供體心臟的低溫安全保存期目前僅有4～6小時,極大地限制了心臟移植的開展。因此,如何有效地延長離體心臟安全保存期是心臟移植中亟待解決的問題,而闡明心肌冷缺血損傷的病理生理學(xué)機(jī)制,尋找更有效的抗心肌冷缺血/再灌注損傷措施則是一個重大的研究課題。細(xì)胞骨架是細(xì)胞內(nèi)重要的保守結(jié)構(gòu),在維持細(xì)胞內(nèi)外的有序性空間結(jié)構(gòu)和細(xì)胞的生命活動中起重要作用。由于細(xì)胞骨架的完整性與溫度有密切關(guān)系,因此,研究低溫環(huán)境下心肌細(xì)胞骨架的變化對細(xì)胞結(jié)構(gòu)和功能的影響十分必要。該課題依據(jù)近年來國內(nèi)外在細(xì)胞骨架研究方面所取得的新進(jìn)展,從細(xì)胞的結(jié)構(gòu)與功能關(guān)系的角度出發(fā),探討低溫環(huán)境下心肌細(xì)胞骨架微管與心肌冷缺血損傷的關(guān)系,以及將保護(hù)細(xì)胞骨架作為新的心肌保護(hù)措施的可能性。該研究對于解決器官移植中供體心臟的安全保存難題有重要意義。 本研究分三部分進(jìn)行。第一部分進(jìn)行成年大鼠離體心臟低溫保存和復(fù)溫再灌注實驗,在保存液中加入微管穩(wěn)定劑和解聚劑,經(jīng)過6小時低溫保存后,復(fù)溫再灌注,檢測冠脈漏出液中心肌酶的含量,同時進(jìn)行形態(tài)學(xué)觀察,探討微管的解聚與穩(wěn)定對離體心臟低溫保存的影響。 第二部分,進(jìn)行成年大鼠分離心肌細(xì)胞低溫保存實驗,分別利用組化免疫熒光和生物化學(xué)比色分析的方法,觀察心肌細(xì)胞經(jīng)歷低溫保存后微管的變化和心肌酶釋放率以及心肌細(xì)胞存活率,探討微管的改變與心肌細(xì)胞損傷之間的關(guān)系。 第三部分,觀察微管穩(wěn)定劑對低溫保存和復(fù)溫再灌注大鼠心肌細(xì)胞凋亡發(fā)生的影響;進(jìn)一步說明細(xì)胞骨架與心肌細(xì)胞凋亡發(fā)生的關(guān)系。 實驗方法 1、離體鼠心灌流和低溫保存 SD大鼠隨機(jī)分成3組,包括對照組(HTK液保存),紫杉酚組(HTK液中含紫杉酚,濃度為10~(-6)M)和秋水仙素組(HTK液中含秋水仙素,濃度為10~(-6)M)。動物麻醉,氣管插管,呼吸機(jī)輔助呼吸,主動脈插入灌注管并迅速取下心臟,在37℃恒溫條件下利用離體鼠心Langendorff灌注模式裝置,進(jìn)行常規(guī)逆行恒壓有氧灌注,待心肌血流動力學(xué)穩(wěn)定后,留取冠脈漏出液,改為經(jīng)主動脈灌注不同的心肌保存液,心臟停跳,放入4℃不同保存液中保存6h,復(fù)溫再灌時留取冠脈漏出液,并切取心肌組織做光鏡和電鏡觀察。 2、成年SD大鼠心肌細(xì)胞分離 灌流方式同前,但是分別用有鈣臺氏液和無鈣臺氏液灌流。再用含Ⅱ型膠原酶和小牛血清白蛋白(BSA)的無鈣臺氏液灌注并消化心臟。然后再用KB液沖洗殘酶。灌流過程持續(xù)充氧飽和。剪碎心室部分,輕輕振蕩,使細(xì)胞分離,然后濾過、低速離心、傾出上清液(含部分死亡破碎的細(xì)胞),即得到實驗用細(xì)胞,換KB液保存。 3、分離心肌細(xì)胞處理方法 將每一次分離得到的心肌細(xì)胞隨機(jī)分為對照組和實驗組,各組設(shè)4℃保存0.5h和3h。實驗組添加紫杉酚(最終濃度為10~(-6)M)。分別在相應(yīng)的保存時間點(diǎn)取出,吸取細(xì)胞懸液涂片,進(jìn)行微管免疫熒光檢測和臺盼藍(lán)染色,余下懸液經(jīng)過高速離心后,移出上清液,做LDH活力分析,沉淀細(xì)胞用同體積的含1%Triton X-100(Sigma)的PBS 37℃下孵育30min后做LDH活力分析。媒介中(釋放出的)和細(xì)胞溶解(保留的)的LDH活力,通過分光光度計和試劑盒檢測出來。LDH釋放以占總LDH活力(釋放+保留)的百分比表示。 4、免疫熒光染色方法 分離的單細(xì)胞涂片,涼干,然后在冷(-20℃)甲醇中固定3min,再用冷丙酮浸濕三次。放入冰箱,4℃保存,第二日做微管免疫熒光檢測。熒光顯微鏡下觀察拍照,圖像經(jīng)過Image-Pro Plus圖象分析軟件(Media cybernetics公司)分析檢測。 5、臺盼藍(lán)染色 涂片后用0.1%臺盼藍(lán)滴染,高倍鏡下(×400)計數(shù)200個細(xì)胞,計算拒染的桿形細(xì)胞所占的比率。 6、凋亡檢測 采用Roche公司檢測試劑盒,運(yùn)用原位末端脫氧核苷酸轉(zhuǎn)移酶介導(dǎo)的熒光素脫氧尿嘧啶核苷酸缺口末端標(biāo)記法(TUNEL)進(jìn)行凋亡細(xì)胞檢測,結(jié)果判定;TUNEL陽性細(xì)胞核呈現(xiàn)紫色或紫黑色,陰性對照和正常細(xì)胞核呈淡粉色。 7、漏出心肌酶檢測 采用試劑盒對離體心臟保存前和再灌注后的冠脈漏出液以及分離心肌細(xì)胞低溫保存后釋放出的和細(xì)胞內(nèi)殘留的心肌酶進(jìn)行生物化學(xué)比色分析。 8、光鏡和電鏡觀察 對心臟低溫保存和復(fù)溫再灌注后的心肌進(jìn)行常規(guī)形態(tài)和凋亡觀察。 實驗結(jié)果 1、分離的成年大鼠心肌細(xì)胞低溫下微管的改變 低溫保存30min后,細(xì)胞微管結(jié)構(gòu)即遭到解聚,表現(xiàn)為免疫熒光減弱,微管結(jié)構(gòu)的連續(xù)性開始喪失,變得粗糙,不光滑,微管結(jié)構(gòu)不清晰。對照組(單純KB液保存)變化明顯突出,保存05.h后微管平均光密度為20.20±1.49,3h后為12.46±1.66;而實驗組(含紫杉酚)變化輕于對照組,保存0.5h后為25.49±1.52,3h后為20.41±1.52,在相同保存時間點(diǎn)實驗組與對照組比較P＜0.05。隨著保存時間延長,兩組微管熒光逐漸減弱,微管網(wǎng)絡(luò)進(jìn)行性丟失。 2、分離的成年大鼠心肌細(xì)胞低溫下LDH釋放率 乳酸脫氫酶(LDH)從保存的心肌細(xì)胞中釋放的情況,在經(jīng)過不同保存時間后被檢測。實驗結(jié)果表明實驗組(保存液含微管穩(wěn)定劑)低溫保存0.5h后LDH釋放率為51.88%±1.06,3h后為58.87%±1.09,明顯少于對照組(保存液不含微管穩(wěn)定劑);對照組低溫保存0.5h后LDH釋放率為56.44%±1.45,3h后為65.68%±1.37。在相同保存時間點(diǎn)實驗組與對照組比較P＜0.05。 3、分離的成年大鼠心肌細(xì)胞低溫下心肌細(xì)胞存活率 細(xì)胞存活率是通過臺盼藍(lán)染色的方法觀測到的,實驗結(jié)果表明在低溫保存3h后兩組差距明顯,對照組細(xì)胞存活率為27.2%±5.05,實驗組為38.80%±2.66。實驗組與對照組比較P＜0.05。 4、成年大鼠心臟低溫保存復(fù)溫再灌注后心肌光鏡下改變 對照組(HTK保存液)心內(nèi)膜下、心肌層、心外膜下均有水腫,表現(xiàn)為細(xì)胞間隙增寬、血管周圍間隙增大,心肌細(xì)胞明顯變細(xì)、皺縮,但心肌細(xì)胞邊緣整齊。紫杉酚組與對照組比較,水腫明顯減輕,心肌細(xì)胞周圍間隙變小,心肌細(xì)胞邊緣整齊、形態(tài)基本正常;秋水仙素組心壁各層明顯水腫,心肌細(xì)胞邊緣不整,細(xì)胞周圍間隙較大,部分心肌細(xì)胞呈結(jié)節(jié)狀溶解。 5、成年大鼠心臟低溫保存復(fù)溫再灌注后心肌超微結(jié)構(gòu)改變 對照組線粒體腫脹,嵴結(jié)構(gòu)部分溶解,肌質(zhì)管擴(kuò)張,但肌節(jié)結(jié)構(gòu)仍然清楚,肌絲結(jié)構(gòu)清晰、無溶解。紫杉酚組無線粒體腫脹,嵴結(jié)構(gòu)正常,肌原纖維排列整齊,肌節(jié)各帶清晰可見,肌質(zhì)管擴(kuò)張不明顯。秋水仙素組肌原纖維結(jié)構(gòu)紊亂,肌節(jié)各帶結(jié)構(gòu)不完整,線粒體明顯擴(kuò)張,線粒體嵴溶解、空化現(xiàn)象嚴(yán)重,肌質(zhì)管明顯擴(kuò)張,肌膜腫脹,呈指狀突起。 6、成年大鼠心臟低溫保存復(fù)溫再灌注后冠脈漏出液中心肌酶的檢測 保存后復(fù)灌4min時各組心肌酶(LDH,GOT,CK)漏出量以秋水仙素組最多,紫杉酚組最少,對照組次之,三組間差異顯著。 7、成年大鼠心臟低溫保存和復(fù)溫再灌注后心肌細(xì)胞凋亡的發(fā)生 在各對照組和實驗組內(nèi),電鏡下觀察都可見到心肌細(xì)胞凋亡的核像改變,部分核的染色質(zhì)向核膜集聚,部分核的染色質(zhì)積聚成塊,不均勻分布。 原位末端標(biāo)記(TUNEL法)的切片鏡下觀察可見,發(fā)生凋亡的細(xì)胞主要是心肌細(xì)胞,少數(shù)為浸潤的淋巴細(xì)胞和血管內(nèi)皮細(xì)胞。組內(nèi)比較和組間比較,結(jié)果;(1)在對照組和實驗組內(nèi),離體鼠心低溫保存時間與凋亡發(fā)生指數(shù)成正比,低溫保存時間越長凋亡細(xì)胞越多(P＜0.01),6h鼠心低溫保存的心肌凋亡細(xì)胞數(shù)量明顯多于4h低溫保存的心肌凋亡細(xì)胞數(shù)量;(2)在對照組和實驗組內(nèi),低溫保存時間越長,對復(fù)溫再灌注后的凋亡細(xì)胞發(fā)生影響也越大,凋亡細(xì)胞數(shù)量越多,即6h低溫保存復(fù)溫再灌注后的凋亡細(xì)胞數(shù)量明顯多于4h低溫保存復(fù)溫再灌注后的心肌凋亡細(xì)胞數(shù)量(P＜0.01);(3)在實驗組和對照組內(nèi),復(fù)溫再灌注后的凋亡細(xì)胞指數(shù)明顯高于再灌注前的單純低溫保存的凋亡細(xì)胞指數(shù)(P＜0.01),可見復(fù)溫再灌注加重了細(xì)胞損傷,增加了心肌細(xì)胞凋亡數(shù)量;(4)實驗組與對照組相比,在各個保存和再灌注時間點(diǎn)上,凋亡指數(shù)都明顯低于對照組(P＜0.01),含有紫杉酚(10~(-6)M)的供心保存液THK液具有明顯的低溫供心保護(hù)作用。 結(jié)論 1、4℃低溫下,分離心肌細(xì)胞保存0.5h心肌細(xì)胞微管即發(fā)生解聚,保存3h微管解聚加重,解聚程度與低溫缺血時間成正比。紫杉酚可以穩(wěn)定微管,減輕微管的解聚。 2、4℃低溫下離體心臟心肌細(xì)胞損傷、心肌酶(LDH,GOT,CK)釋放,與微管解聚成正相關(guān),穩(wěn)定微管可以減輕心肌細(xì)胞釋放LDH、GOT和CK。 3、4℃低溫下可以導(dǎo)致離體心臟線粒體結(jié)構(gòu)受損,微管解聚劑可以加重線粒體損害,微管穩(wěn)定劑可以減輕這種損害。 4、4℃低溫保存可以引起離體心臟心肌細(xì)胞凋亡發(fā)生,復(fù)溫再灌注能加重凋亡發(fā)生,低溫保存時間越長,復(fù)溫再灌注后心肌細(xì)胞凋亡發(fā)生的越多。微管穩(wěn)定劑可以減輕低溫保存下心肌細(xì)胞凋亡的發(fā)生。
[Abstract]:Relationship between changes of myocardial cytoskeleton microtubules and cold ischemic injury at low temperature
objective
Heart transplantation is recognized as the only effective means to treat end-stage heart disease. However, the safe preservation period of the donor heart is only 4~6 hours, which greatly restricts the development of heart transplantation. Therefore, how to effectively prolong the safe preservation period of the isolated heart is an urgent problem in the heart transplantation, and to clarify the cold ischemia of the heart. The pathophysiological mechanism of injury and the search for more effective measures against myocardial cold ischemia / reperfusion injury are a major research topic. The cytoskeleton is an important conservative structure within the cell. It plays an important role in maintaining the orderly spatial structure of cells inside and outside the cell and the life activities of the cells. Therefore, it is necessary to study the influence of the changes of the cytoskeleton on the structure and function of the cardiomyocytes in the low temperature environment. This topic is based on the recent progress in the study of cytoskeleton at home and abroad. From the perspective of the relationship between the structure and function of the cells, the framework of cardiac myocyte microtubules and myocardium under low temperature environment is discussed. The relationship between cold ischemic injury and the possibility of protecting the cytoskeleton as a new myocardial protective measure. This study is of great importance to solving the problem of the safe preservation of the donor heart in organ transplantation.
The study is divided into three parts. The first part is the cryopreservation and rewarming reperfusion experiment of the isolated adult rat heart, the microtubule stabilizer and the depolymerization agent are added in the preservation solution. After 6 hours of cryopreservation, the content of the central muscle enzyme in the leakage of the coronary artery is detected, and the morphological observation is carried out to explore the depolymerization and stability of the microtubule. Influence on cryopreservation of isolated heart.
In the second part, the cryopreservation experiments of isolated adult rat cardiomyocytes were carried out. The changes in microtubule, the rate of myocardial enzyme release and the survival rate of myocardial cells after cryopreservation were observed by the histochemical immunofluorescence and biochemical colorimetric analysis. The relationship between the change of microtubule and the injury of cardiac myocytes was investigated.
The third part was to observe the effect of microtubule stabilizers on the apoptosis of myocardial cells in rats with cryopreservation and rewarming reperfusion, and further explain the relationship between the cytoskeleton and the occurrence of cardiomyocyte apoptosis.
Experimental method
1, isolated rat heart perfusion and cryopreservation
SD rats were randomly divided into 3 groups, including the control group (HTK solution), paclitaxol group (paclitaxol in HTK solution, concentration of 10~ (-6) M) and colchicine group (HTK solution containing colchicine and 10~ (-6) M). Animal anesthesia, tracheal intubation, ventilator assisted respiration, aorta inserted into the perfusion tube and quickly removed the heart, and were used at the constant temperature of 37 degrees. The body rat heart Langendorff perfusion model was used to perform conventional retrograde constant pressure aerobic perfusion. After the myocardial hemodynamic stability was stable, the coronary leaks were retained and changed into the aorta perfusion of different myocardial preservation solution, the heart was stopped, the 6h was stored in the different preservation solution at 4 degrees C, and the coronary leaks were retained in the rewarm reperfusion, and the myocardial tissue was cut into the light microscope. And electron microscope observation.
2, isolation of cardiac myocytes from adult SD rats
The perfusion method was in the same way, but was perfused with calcium table and no calcium table. Then the heart was perfused with a calcium free group of collagenase and calf serum albumin (BSA). Then KB solution was used to rinse the residual enzyme. The perfusion process continued to be saturated with oxygen. Heart, pour out the supernatant (containing some dead and broken cells), that is, get the experimental cells and save it with KB solution.
3, isolation of cardiac myocytes
The isolated myocardial cells were randomly divided into the control group and the experimental group. Each group was set up at 4 C for 0.5h and 3h. experimental group to add paclitaxel (the final concentration of 10~ (-6) M). The cells were removed at the corresponding storage time, and the cell suspension smears were taken respectively. The microtubule immunofluorescence detection and trypan blue staining were carried out. The remaining suspension was centrifuged after high speed centrifugation. When the supernatant was removed and the LDH activity was analyzed, the precipitated cells were incubated with the same volume of 1%Triton X-100 (Sigma) at PBS 37 C for LDH activity analysis. The LDH activity in the medium (released) and the cell dissolution (retained) was detected by the spectrophotometer and the reagent box to determine the percentage of.LDH release to account for the percentage of total LDH activity (release + retention). Express.
4, immunofluorescence staining method
Separate single cell smear, cool dry, then immobilized 3min in cold (-20) methanol, and soak three times with cold acetone. Put it in the refrigerator, save at 4, and do microtubule immunofluorescence for second days. A fluorescence microscope is used to observe and take pictures. The image is analyzed by Image-Pro Plus image analysis software (Media cybernetics company).
5, trypan blue staining
After smear, 0.1% trypan blue drops were stained, and 200 cells were counted at high magnification (x 400), and the percentage of rod cells was calculated.
6, apoptosis detection
The Roche detection kit was used to detect the apoptotic cells using in situ terminal deoxynucleotidyl transferase mediated DD Nick Nick nick end labeling (TUNEL). The results were determined, and the TUNEL positive nuclei were purple or purple black, and the negative control and normal nucleus were pale pink.
7, leakage of myocardial enzyme detection
Biochemical colorimetric analysis was performed on the myocardial enzymes that were released before and after the cryopreservation of isolated myocardial cells and after the cryopreservation of isolated cardiac cells, and the residual myocardial enzymes were released before and after the preservation of the isolated heart.
8, observation of light and electron microscope
The morphology and apoptosis of myocardium after cryopreservation and rewarming were observed.
experimental result
1, changes in microtubules of isolated adult rat cardiomyocytes at low temperatures.
After cryopreservation of 30min, the cell microtubule structure was depolymerization, which showed that the immunofluorescence was weakened, the continuity of microtubule structure began to lose, became rough, not smooth, and the microtubule structure was not clear. The change of the control group (pure KB solution) was obviously prominent, and the average optical density of microtubule after 05.h was 20.20 + 1.49,3h after 12.46 + 1.66; and the experimental group (including the experimental group) The change of paclitaxel was lighter than that of the control group. After the preservation of 0.5h, it was 25.49 + 1.52,3h after 20.41 + 1.52. In the same storage time, the experimental group was compared with the control group. The P < 0.05. decreased with the prolongation of the preservation time. The microtubule fluorescence in the two groups gradually weakened, and the microtubule network was lost.
2, the release rate of LDH from isolated adult rat cardiomyocytes at low temperature.
The release of lactate dehydrogenase (LDH) from preserved cardiac myocytes was detected after different preservation time. The experimental results showed that the experimental group (preservation solution containing microtubule stabilizer) was 58.87% + 1.09 after the cryopreservation of 0.5h, and the LDH release rate was 51.88% + 1.06,3h, obviously less than that in the control group (the preservative without microtubule stabilizer); the control group was preserved at low temperature. The release rate of LDH after 0.5h storage was 56.44% + 1.45,3h and 65.68% + 1.37. at the same storage time. The experimental group was compared with the control group P < 0.05.
3, the viability of isolated adult rat cardiomyocytes at low temperatures.
The cell survival rate was observed by trypan blue staining. The experimental results showed that the two groups in the two groups were significantly different after cryopreservation, and the cell survival rate of the control group was 27.2% + 5.05, the experimental group was 38.80% + 2.66. and the control group was P < 0.05.
4, the myocardium of adult rats was changed by cryopreservation and reperfusion.
In the control group (HTK preservation solution), there was edema under the endocardium, myocardial layer and epicardium, showing the widening of the intercellular space, the enlargement of the perivascular space, the obvious thinning and crinkling of cardiac myocytes, but the edge of the cardiac myocytes was neat. In normal colchicine group, the wall of heart wall was obviously edema, the edge of cardiac myocytes was irregular, the surrounding cells were larger, and some myocardial cells were nodular dissolved.
5, the ultrastructural changes of myocardium in adult rats after hypothermic preservation and rewarming and reperfusion
In the control group, the mitochondria were swollen, the ridge structure was partially dissolved, the muscle tube expanded, but the myofibrillar structure was clear, the myofibrous structure was clear and insoluble. The paclitaxel group was swollen, the ridge structure was normal, the myofibrils were arranged neatly, the myofibrils were clearly visible and the myofibrillar expansion was not obvious. The colchicine group myofibrillar structure disorder, the myofibrils were all band knot. The structure is incomplete, mitochondria are obviously dilated, mitochondria cristae are dissolved, cavitation is serious, muscular tube is obviously dilated, muscular membrane is swollen, and finger like protuberance.
6, the detection of myocardial enzymes in the coronary leakage of adult rats after hypothermic preservation and rewarming and reperfusion
The amount of myocardial enzyme (LDH, GOT, CK) leakage in each group was the largest in the colchicine group after storage and reperfusion at 4min. The paclitaxel group was the least and the control group was the second. There was a significant difference between the three groups.
7, cardiomyocyte apoptosis occurred after cryopreservation and rewarming in adult rats.
In the control group and the experimental group, the nuclear image of the cardiomyocyte apoptosis could be observed under the electron microscope, the chromatin of some nuclei gathered into the nuclear membrane, and the chromatin of some nuclei accumulated into block and uneven distribution.
In situ end labeling (TUNEL) microscopic examination revealed that the apoptotic cells were mainly cardiac myocytes, few of which were infiltrated lymphocytes and vascular endothelial cells. Comparison and comparison between groups were compared, and the results were compared. (1) in the control group and the experimental group, the cryopreservation time of the isolated rat heart was proportional to the apoptotic index, and the longer the cryopreservation time was. The more long apoptotic cells (P < 0.01), the number of apoptotic cells preserved at low temperature in 6h rat heart is more than that of 4H cryopreserved apoptotic cells. (2) the longer the cryopreservation time in the control group and the experimental group, the greater the effect on the apoptotic cells after rewarming and reperfusion, the more the number of apoptotic cells, that is, the cryopreservation and rewarming of the 6h at low temperature. The number of apoptotic cells after perfusion was significantly more than that of 4H at low temperature and reperfusion (P < 0.01). (3) in the experimental and control groups, the apoptotic cell index after rewarming and reperfusion was significantly higher than that of the apoptotic cell index (P < 0.01), which was preserved at low temperature before reperfusion (P < 0.01). The number of cardiomyocytes apoptosis increased. (4) compared with the control group, the apoptosis index of the experimental group was significantly lower than the control group (P < 0.01), and the THK solution containing paclitaxel (10~ (-6) M) had a significant protective effect on the cryogenic donor heart.
conclusion
At 1,4 C, the microtubule of 0.5h cardiomyocytes was depolymerization, and the depolymerization of 3H microtubules was aggravated. The degree of depolymerization was proportional to the time of low temperature ischemia. Taxol could stabilize microtubules and reduce the depolymerization of microtubules.
The release of myocardial enzymes (LDH, GOT, CK) in isolated cardiac myocytes at 2,4 C at low temperature is positively related to the depolymerization of microtubules. Stable microtubules can reduce the release of LDH, GOT and CK. in cardiac myocytes.
Low temperature at 3,4 C can lead to impaired mitochondrial structure in isolated heart. Microtubule depolymerization agent can aggravate mitochondrial damage. Microtubule stabilizer can alleviate this damage.
The cryopreservation of 4,4 C can cause apoptosis in cardiac myocyte in vitro. Rewarming reperfusion can aggravate apoptosis. The longer the cryopreservation time is, the more apoptosis occurs in myocardial cells after rewarming and reperfusion. Microtubule stabilizer can reduce the occurrence of cardiomyocyte apoptosis under cryopreservation.

【學(xué)位授予單位】：中國醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2007
【分類號】：R363

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 張晶;黃芪甲苷對后負(fù)荷過載型心肌肥厚的抑制作用與機(jī)制研究[D];遼寧醫(yī)學(xué)院;2012年

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本文編號：1894472