中国药物与临床
中國藥物與臨床
중국약물여림상
CHINESE REMEDIES & CLINICS
2015年
8期
1064-1067
,共4页
沉默信息调节蛋白质类%细胞衰老%半乳糖
沉默信息調節蛋白質類%細胞衰老%半乳糖
침묵신식조절단백질류%세포쇠로%반유당
Silent information regulator proteins%Cell aging%Galactose
目的：通过观察SIRT1全长（SIRT1-FL）及缺失第8外显子的SIRT1选择性剪接变异体（SIRT1-ΔExon8）在D-半乳糖诱导的293T细胞衰老模型中的表达变化，初步探讨SIRT1全长及SIRT1-ΔExon8对细胞衰老进程的影响。方法①细胞计数kit-8（CCK8）法测定不同浓度D-半乳糖（0，5，10，15，20 g/L）对293T细胞活力的影响。②用含10 g/L的D-半乳糖达氏修正伊氏培养基作用于第3代293T细胞，继续培养至第6代，建立细胞衰老模型，β-半乳糖苷酶（SA-β-gal）染色观察衰老细胞阳性率。③实时聚合酶链反应（RT-PCR）检测诱导组与对照组细胞中SIRT1-FL及SIRT1-ΔExon8 mRNA的表达水平。结果①CCK-8结果显示10 g/L的D-半乳糖能诱导细胞衰老且不引起细胞过度损伤。②诱导组衰老细胞阳性率（74.2±3.6）％高于对照组（7.7±1.4）％（P＜0.05）。③RT-PCR结果显示诱导组SIRT1-FL和SIRT1-ΔExon8 mRNA表达水平分别较对照组降低了（78.26±0.05）％和（88.03±0.03）％（P＜0.05）。结论①SA-β-gal染色结果提示D-半乳糖诱导293T细胞衰老模型建立成功。②衰老细胞组的SIRT1-FL和SIRT1-ΔExon8mRNA表达水平较对照组均显著降低，提示SIRT1-FL及SIRT1-ΔExon8可能参与调控细胞衰老的进程。
目的：通過觀察SIRT1全長（SIRT1-FL）及缺失第8外顯子的SIRT1選擇性剪接變異體（SIRT1-ΔExon8）在D-半乳糖誘導的293T細胞衰老模型中的錶達變化，初步探討SIRT1全長及SIRT1-ΔExon8對細胞衰老進程的影響。方法①細胞計數kit-8（CCK8）法測定不同濃度D-半乳糖（0，5，10，15，20 g/L）對293T細胞活力的影響。②用含10 g/L的D-半乳糖達氏脩正伊氏培養基作用于第3代293T細胞，繼續培養至第6代，建立細胞衰老模型，β-半乳糖苷酶（SA-β-gal）染色觀察衰老細胞暘性率。③實時聚閤酶鏈反應（RT-PCR）檢測誘導組與對照組細胞中SIRT1-FL及SIRT1-ΔExon8 mRNA的錶達水平。結果①CCK-8結果顯示10 g/L的D-半乳糖能誘導細胞衰老且不引起細胞過度損傷。②誘導組衰老細胞暘性率（74.2±3.6）％高于對照組（7.7±1.4）％（P＜0.05）。③RT-PCR結果顯示誘導組SIRT1-FL和SIRT1-ΔExon8 mRNA錶達水平分彆較對照組降低瞭（78.26±0.05）％和（88.03±0.03）％（P＜0.05）。結論①SA-β-gal染色結果提示D-半乳糖誘導293T細胞衰老模型建立成功。②衰老細胞組的SIRT1-FL和SIRT1-ΔExon8mRNA錶達水平較對照組均顯著降低，提示SIRT1-FL及SIRT1-ΔExon8可能參與調控細胞衰老的進程。
목적：통과관찰SIRT1전장（SIRT1-FL）급결실제8외현자적SIRT1선택성전접변이체（SIRT1-ΔExon8）재D-반유당유도적293T세포쇠로모형중적표체변화，초보탐토SIRT1전장급SIRT1-ΔExon8대세포쇠로진정적영향。방법①세포계수kit-8（CCK8）법측정불동농도D-반유당（0，5，10，15，20 g/L）대293T세포활력적영향。②용함10 g/L적D-반유당체씨수정이씨배양기작용우제3대293T세포，계속배양지제6대，건립세포쇠로모형，β-반유당감매（SA-β-gal）염색관찰쇠로세포양성솔。③실시취합매련반응（RT-PCR）검측유도조여대조조세포중SIRT1-FL급SIRT1-ΔExon8 mRNA적표체수평。결과①CCK-8결과현시10 g/L적D-반유당능유도세포쇠로차불인기세포과도손상。②유도조쇠로세포양성솔（74.2±3.6）％고우대조조（7.7±1.4）％（P＜0.05）。③RT-PCR결과현시유도조SIRT1-FL화SIRT1-ΔExon8 mRNA표체수평분별교대조조강저료（78.26±0.05）％화（88.03±0.03）％（P＜0.05）。결론①SA-β-gal염색결과제시D-반유당유도293T세포쇠로모형건립성공。②쇠로세포조적SIRT1-FL화SIRT1-ΔExon8mRNA표체수평교대조조균현저강저，제시SIRT1-FL급SIRT1-ΔExon8가능삼여조공세포쇠로적진정。
Objective To investigate the effect of silent mating type information regulation 2 homolog 1-full long (SIRT1-FL) and its alternative splicing variant SIRT1-ΔExon8 on the aging process of 293T cells by observing the expression changes of SIRT1 and SIRT1-ΔExon8 in the D-galactose (D-gal) induced senescence model of 293T cells. Methods ①Cell Counting Kit-8 (CCK8) was used to determine the effect of D-gal on 293T cells viability at different concentrations (0, 5, 10, 15, 20 g/L). ②The 293T cells were cultured in DMEM high-glucose medium con-taining 10 g/L D-gal at the third passage and amplified to the sixth passage, and the senescence model of the 293T cells was established. β-galactosidase (SA-β-Gal) staining was used to determine the positive rate of senescent cells.③The mRNA expression levels of SIRT1 and SIRT1-ΔExon8 were measured in the 293T cells of the D-gal induced group and the control group by RT-PCR assay. Results ①The CCK-8 assay showed that the 10 g/L D-gal could in-duce 293T cells senescence and avoid extensive injuries. ②The positive rate of senescent cells in the D-gal induced group (74.2±3.6)%was higher than that in the control group (7.7±1.4)% (P<0.05). ③The findings of RT-PCR showed that the mRNA expression levels of SIRT1-FL and SIRT1-ΔExon8 in the 293T cells in the D-gal induced group were decreased by (78.26±0.05)%and (88.03±0.03)%compared with those in the control group, respectively (P<0.05). Conclusion ①SA-β-gal staining shows that the D-gal can induce the senescence model of 293T cells successfully.②The mRNA expression levels of SIRT1-FL and SIRT1-ΔExon8 in the senescence cells group are significantly de-creased compared with those in the control group, suggesting SIRT1-FL and SIRT1-ΔExon8 may be involved in the regulation of 293T cells senescence.
