中国临床医学
中國臨床醫學
중국림상의학
Chinese Journal of Clinical Medicine
2015年
4期
479-481
,共3页
张亚平%颜彦%宋振举%童朝阳
張亞平%顏彥%宋振舉%童朝暘
장아평%안언%송진거%동조양
PRKAG2 心脏综合征%R302Q 突变%糖原%钙超载
PRKAG2 心髒綜閤徵%R302Q 突變%糖原%鈣超載
PRKAG2 심장종합정%R302Q 돌변%당원%개초재
PRKAG2 cardiac syndrome%R302Q mutation%Glycogen%Calcium overload
目的::探讨 PRKAG2心脏综合征中 R302Q 突变对心肌细胞糖原和钙离子稳态的影响。方法:用表达增强型绿色荧光蛋白(enhanced green fluorescent protein,EGFP)、PRKAG2-WT、PRKAG2-R302Q 的腺病毒感染大鼠 H9C2心肌细胞,同时设正常对照组。感染24 h 和48 h 后,采用过碘酸–雪夫(Periodic acid-Schiff,PAS)染色法进行糖原染色,观察心肌细胞的糖原贮积;采用酶联免疫吸附试验(ELISA)法检测细胞内糖原含量;同时用 Rohd-2/AM 活体染料进行钙离子染色,然后采用流式细胞术分析钙离子库信号差异。结果:EGFP 组与对照组糖原染色和含量结果无明显差别;PRKAG2-WT 组糖原染色和含量略微增强;PRKAG2-R302Q 组糖原染色和含量明显增强。各组的钙离子稳态并未受到影响。结论:PRKAG2心脏综合征中R302Q 突变导致心肌细胞内糖原贮积,而钙离子稳态并未受到影响,没有钙超载现象发生。
目的::探討 PRKAG2心髒綜閤徵中 R302Q 突變對心肌細胞糖原和鈣離子穩態的影響。方法:用錶達增彊型綠色熒光蛋白(enhanced green fluorescent protein,EGFP)、PRKAG2-WT、PRKAG2-R302Q 的腺病毒感染大鼠 H9C2心肌細胞,同時設正常對照組。感染24 h 和48 h 後,採用過碘痠–雪伕(Periodic acid-Schiff,PAS)染色法進行糖原染色,觀察心肌細胞的糖原貯積;採用酶聯免疫吸附試驗(ELISA)法檢測細胞內糖原含量;同時用 Rohd-2/AM 活體染料進行鈣離子染色,然後採用流式細胞術分析鈣離子庫信號差異。結果:EGFP 組與對照組糖原染色和含量結果無明顯差彆;PRKAG2-WT 組糖原染色和含量略微增彊;PRKAG2-R302Q 組糖原染色和含量明顯增彊。各組的鈣離子穩態併未受到影響。結論:PRKAG2心髒綜閤徵中R302Q 突變導緻心肌細胞內糖原貯積,而鈣離子穩態併未受到影響,沒有鈣超載現象髮生。
목적::탐토 PRKAG2심장종합정중 R302Q 돌변대심기세포당원화개리자은태적영향。방법:용표체증강형록색형광단백(enhanced green fluorescent protein,EGFP)、PRKAG2-WT、PRKAG2-R302Q 적선병독감염대서 H9C2심기세포,동시설정상대조조。감염24 h 화48 h 후,채용과전산–설부(Periodic acid-Schiff,PAS)염색법진행당원염색,관찰심기세포적당원저적;채용매련면역흡부시험(ELISA)법검측세포내당원함량;동시용 Rohd-2/AM 활체염료진행개리자염색,연후채용류식세포술분석개리자고신호차이。결과:EGFP 조여대조조당원염색화함량결과무명현차별;PRKAG2-WT 조당원염색화함량략미증강;PRKAG2-R302Q 조당원염색화함량명현증강。각조적개리자은태병미수도영향。결론:PRKAG2심장종합정중R302Q 돌변도치심기세포내당원저적,이개리자은태병미수도영향,몰유개초재현상발생。
Objective:To investigate the effect of R302Q mutation on glycogen and calcium ion homeostasis of cardiomyocytes in PRKAG2 cardiac syndrome.Methods:The rat H9C2 cardiomyocytes were infected with adenovirus containing enhanced green fluorescent protein(EGFP),PRKAG2-WT,and PRKAG2-R302Q,meanwhile,normal control group was set up.After 24 h and 48 h,Periodic acid-Schiff(PAS)staining was used to observe cardiomyocyte glycogen storage,and the glycogen con-tent in cells was detected by enzyme-linked immunosorbent assay (ELISA).Meanwhile,Rohd-2/AM vital dye was used to car-ry out calcium ion staining,and then the signal differences among calcium stores were analyzed by flow cytometry.Results:There was no obvious difference regarding glycogen staining and content between EGFP group and control group.Glycogen staining and content was slightly enhanced in PRKAG2-WT group,while glycogen staining and content was obviously enhanced in PRKAG2-R302Q group.However,the calcium ion homeostasis in each group was not affected.Conclusions:R302Q muta-tion leads to intra cardiomyocyte glycogen accumulation in PRKAG2 cardiac syndrome.However,calcium ion homeostasis is not affected and there is no calcium overload.