脂聯(lián)素對高糖環(huán)境下人心肌細胞凋亡及Bax、Bcl-2表達的影響
發(fā)布時間：2018-11-27 18:16
【摘要】：目的：通過培養(yǎng)人心肌細胞株，觀察高糖環(huán)境下及加入外源脂聯(lián)素干預后不同時間點，人心肌細胞的增殖、凋亡情況，及Bax\Bcl-2 mRNA的表達變化，從而揭示脂聯(lián)素對高糖環(huán)境下心肌細胞的保護作用，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)的人心肌細胞（HCM）隨機分為正常對照組（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運用四甲基偶氮唑藍（MTT）法測定不同時間點各組細胞的光吸收值（OD值），檢測高糖及脂聯(lián)素對人心肌細胞增殖的影響；③收集各個時間段培養(yǎng)的人心肌細胞，離心后加入脂結(jié)合蛋白Annexin V/碘化丙啶（PI）染色后，運用流式細胞儀（FCM）測定各組細胞在不同時間段的凋亡率；④在各個時間段提取各組人心肌細胞的RNA，逆轉(zhuǎn)錄為cDNA，然后加入引物，通過實時熒光定量PCR（RT-PCR）技術(shù)測定各組人心肌細胞Bax、Bcl-2的表達情況。 結(jié)果：①與正常對照組相比，高糖的持續(xù)作用抑制人心肌細胞增殖，而脂聯(lián)素組人心肌細胞的增殖明顯增加，差異有統(tǒng)計學意義（P0.05）； ②高糖組與對照組相比，明顯增加了人心肌細胞的凋亡率，加入脂聯(lián)素干預后，人心肌細胞的凋亡率明顯下降，且差異有統(tǒng)計學意義（P0.05）； ③高糖組人心肌細胞Bax的表達上調(diào)、Bcl-2的表達下降；與對照組相比，差異有統(tǒng)計學意義。脂聯(lián)素組人心肌細胞Bax的表達減少、Bcl-2的表達增加，且該作用呈時間依賴性，與高糖組相比，差異有統(tǒng)計學意義（P0.05）。 結(jié)論：高糖環(huán)境下，人心肌細胞的增殖減少，凋亡率增加，Bax的表達增加，Bcl-2表達降低；而加入脂聯(lián)素干預后，通過下調(diào)促進凋亡因子Bax的表達、上調(diào)抑制凋亡因子Bcl-2的表達，增加細胞的增殖，減少細胞的凋亡，從而保護高糖環(huán)境下人心肌細胞的生長，，發(fā)揮對糖尿病心肌病的保護作用，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.
【學位授予單位】：山西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R363
本文編號：2361655
[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.
【學位授予單位】：山西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R363
【參考文獻】
相關期刊論文 前2條
1 李舒帆;李興;;脂聯(lián)素對高糖環(huán)境下人心肌細胞增生及caspase-3、T-鈣黏蛋白表達的影響[J];醫(yī)學研究雜志;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系[J];中國糖尿病雜志;2008年08期
本文編號：2361655
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