中国药理学与毒理学杂志
中國藥理學與毒理學雜誌
중국약이학여독이학잡지
CHINESE JOURNAL OF PHARMACOLOGY AND TOXICOLOGY
2011年
1期
1-6
,共6页
徐道华%杨玮%周晨慧%刘钰瑜%许碧连
徐道華%楊瑋%週晨慧%劉鈺瑜%許碧連
서도화%양위%주신혜%류옥유%허벽련
间质干细胞%脂细胞%骨质疏松%小檗碱%细胞分化
間質榦細胞%脂細胞%骨質疏鬆%小檗堿%細胞分化
간질간세포%지세포%골질소송%소벽감%세포분화
mesenchymal stem cell%adipocyte%osteoporosis%berberine%cell differentiation
目的 探讨小檗碱对大鼠骨髓间质干细胞成脂分化的影响及其机制.方法 经分离纯化的大鼠骨髓间质干细胞,分为正常对照组,成脂分化诱导液(AIM)模型组及AIM+小檗碱0.1,0.3,1和3 μmol·L-1组.倒置显微镜下观察细胞的形态特征,油红O染色检测脂肪细胞,以对硝基苯磷酸为底物检测碱性磷酸酶(ALP)活性,MTT法检测细胞存活率,RT-PCR检测过氧化物酶体增殖物激活受体γ(PPARγ),脂肪酸结合蛋白(aP2)和CCAAT增强子结合蛋白α(C/EBPα) mRNA表达.结果 与正常对照组比较,AIM模型组细胞形成的脂肪细胞明显增加(P<0.01),ALP活性明显降低(P<0.01),PPARγ,aP2和C/EBPα mRNA表达明显增高(P<0.01).与AIM组比较,AIM+小檗碱显著抑制骨髓间质干细胞成脂分化,小檗碱0.1,0.3,1和3 μmol·L-1显著升高ALP活性,分别增加26%,54%,81%和122%;小檗碱3 μmol·L-1显著下调PPARγ mRNA(0.91±0.10 vs 1.34±0.06),(P<0.01),aP2 mRNA(1.05±0.10 vs 1.53±0.09) (P<0.01)和 C/EBPαmRNA表达(1.24±0.06 vs 1.54±0.09) (P<0.01).小檗碱对骨髓间质干细胞增殖无显著影响.结论 小檗碱能够抑制骨髓间质干细胞成脂分化,该作用可能与其下调PPARγ mRNA,aP2 mRNA和C/EBPα mRNA表达有关.
目的 探討小檗堿對大鼠骨髓間質榦細胞成脂分化的影響及其機製.方法 經分離純化的大鼠骨髓間質榦細胞,分為正常對照組,成脂分化誘導液(AIM)模型組及AIM+小檗堿0.1,0.3,1和3 μmol·L-1組.倒置顯微鏡下觀察細胞的形態特徵,油紅O染色檢測脂肪細胞,以對硝基苯燐痠為底物檢測堿性燐痠酶(ALP)活性,MTT法檢測細胞存活率,RT-PCR檢測過氧化物酶體增殖物激活受體γ(PPARγ),脂肪痠結閤蛋白(aP2)和CCAAT增彊子結閤蛋白α(C/EBPα) mRNA錶達.結果 與正常對照組比較,AIM模型組細胞形成的脂肪細胞明顯增加(P<0.01),ALP活性明顯降低(P<0.01),PPARγ,aP2和C/EBPα mRNA錶達明顯增高(P<0.01).與AIM組比較,AIM+小檗堿顯著抑製骨髓間質榦細胞成脂分化,小檗堿0.1,0.3,1和3 μmol·L-1顯著升高ALP活性,分彆增加26%,54%,81%和122%;小檗堿3 μmol·L-1顯著下調PPARγ mRNA(0.91±0.10 vs 1.34±0.06),(P<0.01),aP2 mRNA(1.05±0.10 vs 1.53±0.09) (P<0.01)和 C/EBPαmRNA錶達(1.24±0.06 vs 1.54±0.09) (P<0.01).小檗堿對骨髓間質榦細胞增殖無顯著影響.結論 小檗堿能夠抑製骨髓間質榦細胞成脂分化,該作用可能與其下調PPARγ mRNA,aP2 mRNA和C/EBPα mRNA錶達有關.
목적 탐토소벽감대대서골수간질간세포성지분화적영향급기궤제.방법 경분리순화적대서골수간질간세포,분위정상대조조,성지분화유도액(AIM)모형조급AIM+소벽감0.1,0.3,1화3 μmol·L-1조.도치현미경하관찰세포적형태특정,유홍O염색검측지방세포,이대초기분린산위저물검측감성린산매(ALP)활성,MTT법검측세포존활솔,RT-PCR검측과양화물매체증식물격활수체γ(PPARγ),지방산결합단백(aP2)화CCAAT증강자결합단백α(C/EBPα) mRNA표체.결과 여정상대조조비교,AIM모형조세포형성적지방세포명현증가(P<0.01),ALP활성명현강저(P<0.01),PPARγ,aP2화C/EBPα mRNA표체명현증고(P<0.01).여AIM조비교,AIM+소벽감현저억제골수간질간세포성지분화,소벽감0.1,0.3,1화3 μmol·L-1현저승고ALP활성,분별증가26%,54%,81%화122%;소벽감3 μmol·L-1현저하조PPARγ mRNA(0.91±0.10 vs 1.34±0.06),(P<0.01),aP2 mRNA(1.05±0.10 vs 1.53±0.09) (P<0.01)화 C/EBPαmRNA표체(1.24±0.06 vs 1.54±0.09) (P<0.01).소벽감대골수간질간세포증식무현저영향.결론 소벽감능구억제골수간질간세포성지분화,해작용가능여기하조PPARγ mRNA,aP2 mRNA화C/EBPα mRNA표체유관.
OBJECTIVE To investigate the effect of berberine on differentiation of rat bone marrow mesenchymal stem cells (MSCs) to adipocytes and its mechanism. METHODS Rat MSCs were isolated and cultured, adipocytic differentiation was induced with adipogenesis-inducing medium (AIM). Cells were assigned into 6 groups:normal control, AIM group, AIM+berberine 0.1, 0.3, 1 and 3 μmol·L-1 groups, respectively. Morphology characteristics of mesenchymal stem cells were observed under an inverted microscope and adipocyte levels were analyzed by oil O staining. Alkaline phosphatase (ALP) activity was detected using p-nitrophenyl phosphate as a substrate. The cell survival was determined by MTT assay. Expressions of peroxisome proliferator activated receptor γ (PPARγ), fatty acid binding protein (aP2) and CCAAT enhancer-binding protein α (C/EBPα) mRNA were detected by semiquantitative RT-PCR. RESULTS Compared with normal control group, MSCs adipogenic differentiation, PPARγ, aP2 and C/EBPα mRNA expression significantly increased in AIM group (P<0.01), ALP activity in AIM group significantly decreased (P<0.01). Compared with AIM group, berberine inhibited MSCs adipogenic differentiation (P<0.01) and berberine 0.1, 0.3, 1 and 3 μmol·L-1 increased ALP activity by 26%, 54%, 81% and 122%, respectively. Berberine 3 μmol·L-1 significantly downregulated PPARγ expression (0.91±0.10 vs 1.34±0.06) (P<0.01), aP2 (1.05±0.10 vs 1.53±0.09) (P<0.01) and C/EBPα mRNA (1.24±0.06 vs 1.54±0.09) (P<0.01). Berberine had no effect on proliferation of MSCs. CONCLUSION Berberine inhibits differentiation of MSCs into adipocytes, which might be closely related to the downregulation of PPARγ, aP2 and C/EBPα mRNA.
