【摘要】：目的：通過(guò)培養(yǎng)人心肌細(xì)胞株，觀察高糖環(huán)境下及加入外源脂聯(lián)素干預(yù)后不同時(shí)間點(diǎn)，人心肌細(xì)胞的增殖、凋亡情況，及Bax\Bcl-2 mRNA的表達(dá)變化，從而揭示脂聯(lián)素對(duì)高糖環(huán)境下心肌細(xì)胞的保護(hù)作用，為臨床糖尿病心肌病的治療提供新的思路。 方法：①將培養(yǎng)的人心肌細(xì)胞（HCM）隨機(jī)分為正常對(duì)照組（D-葡萄糖濃度：5.5mmol/l）、高糖組（D-葡萄糖濃度：30.0mmol/l）、高糖+脂聯(lián)素組（D-葡萄糖濃度：30.0mmol/l+ADPN濃度：2.5μg/ml），按上述條件分別培養(yǎng)24h、48h、72h；②運(yùn)用四甲基偶氮唑藍(lán)（MTT）法測(cè)定不同時(shí)間點(diǎn)各組細(xì)胞的光吸收值（OD值），檢測(cè)高糖及脂聯(lián)素對(duì)人心肌細(xì)胞增殖的影響；③收集各個(gè)時(shí)間段培養(yǎng)的人心肌細(xì)胞，離心后加入脂結(jié)合蛋白Annexin V/碘化丙啶（PI）染色后，運(yùn)用流式細(xì)胞儀（FCM）測(cè)定各組細(xì)胞在不同時(shí)間段的凋亡率；④在各個(gè)時(shí)間段提取各組人心肌細(xì)胞的RNA，逆轉(zhuǎn)錄為cDNA，然后加入引物，通過(guò)實(shí)時(shí)熒光定量PCR（RT-PCR）技術(shù)測(cè)定各組人心肌細(xì)胞Bax、Bcl-2的表達(dá)情況。 結(jié)果：①與正常對(duì)照組相比，高糖的持續(xù)作用抑制人心肌細(xì)胞增殖，而脂聯(lián)素組人心肌細(xì)胞的增殖明顯增加，差異有統(tǒng)計(jì)學(xué)意義（P0.05）； ②高糖組與對(duì)照組相比，明顯增加了人心肌細(xì)胞的凋亡率，加入脂聯(lián)素干預(yù)后，人心肌細(xì)胞的凋亡率明顯下降，且差異有統(tǒng)計(jì)學(xué)意義（P0.05）； ③高糖組人心肌細(xì)胞Bax的表達(dá)上調(diào)、Bcl-2的表達(dá)下降；與對(duì)照組相比，差異有統(tǒng)計(jì)學(xué)意義。脂聯(lián)素組人心肌細(xì)胞Bax的表達(dá)減少、Bcl-2的表達(dá)增加，且該作用呈時(shí)間依賴性，與高糖組相比，差異有統(tǒng)計(jì)學(xué)意義（P0.05）。 結(jié)論：高糖環(huán)境下，人心肌細(xì)胞的增殖減少，凋亡率增加，Bax的表達(dá)增加，Bcl-2表達(dá)降低；而加入脂聯(lián)素干預(yù)后，通過(guò)下調(diào)促進(jìn)凋亡因子Bax的表達(dá)、上調(diào)抑制凋亡因子Bcl-2的表達(dá)，增加細(xì)胞的增殖，減少細(xì)胞的凋亡，從而保護(hù)高糖環(huán)境下人心肌細(xì)胞的生長(zhǎng)，，發(fā)揮對(duì)糖尿病心肌病的保護(hù)作用，為臨床治療糖尿病心肌病提供一條新的途徑。
[Abstract]:Objective: to observe the proliferation, apoptosis and Bax\ Bcl-2 mRNA expression of human cardiomyocytes in high glucose environment and at different time points after exogenous adiponectin (adiponectin). Thus, the protective effect of adiponectin on cardiomyocytes in high glucose environment was revealed, which provided a new idea for the treatment of diabetic cardiomyopathy. Methods: (1) cultured human cardiomyocytes (HCM) were randomly divided into normal control group (D- glucose concentration: 5.5mmol/l) and high glucose group (D- glucose concentration: 30.0mmol/l). High glucose group (D- glucose concentration: 30.0mmol/l ADPN concentration: 2.5 渭 g/ml) was cultured for 24 h and 48 h for 72 h. (2) the light absorption value (OD) of the cells at different time points was measured by (MTT) method, and the effects of high glucose and adiponectin on the proliferation of human cardiomyocytes were detected. 3The cultured human cardiomyocytes were collected. After centrifugation, lipid binding protein (Annexin V) / pyridine iodide (PI) was added, and the apoptosis rate of each group was measured by flow cytometry (FCM) (FCM). 4Recombinant RNA, of human cardiomyocytes was extracted from each group at each time and then primers were added. The expression of Bax,Bcl-2 in human cardiomyocytes was detected by real-time quantitative PCR (RT-PCR) technique. Results: 1Compared with the normal control group, the sustained effect of high glucose inhibited the proliferation of human cardiomyocytes, while the proliferation of human cardiomyocytes in adiponectin group increased significantly (P0.05). 2 compared with the control group, the high glucose group significantly increased the apoptosis rate of human cardiomyocytes. After adiponectin intervention, the apoptosis rate of human cardiomyocytes decreased significantly (P0.05). (3) the expression of Bax was up-regulated and the expression of Bcl-2 was decreased in high glucose group, compared with the control group, the difference was statistically significant. In adiponectin group, the expression of Bax was decreased and the expression of Bcl-2 was increased in a time dependent manner. The difference was statistically significant compared with high glucose group (P0.05). Conclusion: under high glucose condition, the proliferation of human cardiomyocytes decreased, the apoptosis rate increased, the expression of Bax increased and the expression of Bcl-2 decreased. After adiponectin was added, the expression of apoptotic factor Bax was down-regulated, the expression of apoptotic factor Bcl-2 was up-regulated, the proliferation of cells was increased, and the apoptosis was reduced, thus protecting the growth of human cardiomyocytes under high glucose environment. It provides a new way for the clinical treatment of diabetic cardiomyopathy.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R363

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 李舒帆;李興;;脂聯(lián)素對(duì)高糖環(huán)境下人心肌細(xì)胞增生及caspase-3、T-鈣黏蛋白表達(dá)的影響[J];醫(yī)學(xué)研究雜志;2010年06期

2 田臘梅;李興;;脂聯(lián)素及T-鈣黏蛋白與大鼠糖尿病心肌病的關(guān)系[J];中國(guó)糖尿病雜志;2008年08期

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