CAJ | 학술논문

目的：探讨1,25-二羟基维生素D3[1,25(OH)2D3]对人肾小球系膜细胞增殖的影响及其在肾小球系膜细胞调控中对mTOR/p70s6K信号通路的作用。方法体外培养人肾小球系膜细胞，取传代培养至3~7代细胞分为4组：正常对照组（加含5%胎牛血清DMEM培养基），VD组[加1,25(OH)2D310-8 mol/L]，R组（加雷帕霉素5 mg/L），R+VD组[加雷帕霉素5 mg/L及1,25(OH)2D310-8 mol/L]，干预48 h。采用细胞增殖与活性检测试剂盒CCK-8检测细胞增殖情况，流式细胞术检测细胞周期时相分布，免疫荧光法检测细胞中mTOR、p70s6K的表达情况。结果正常对照组、VD组、R组、R+VD组：（1）吸光度值（A450）依次降低，组间多重比较差异均有统计学意义（均P<0.05）；抑制率（IR）依次升高。（2）G1期细胞比例依次增加，S期、G2/M期依次减少，增殖指数（PI）依次降低，除R组与VD组比较差异无统计学意义外，其余组间多重比较差异均有统计学意义。（3）系膜细胞中mTOR、p70s6K蛋白表达强度均依次降低，除R组与VD组比较差异无统计学意义外，其余组间多重比较差异均有统计学意义。结论1,25(OH)2D3可显著抑制人系膜细胞增殖，且其可能通过抑制mTOR/p70s6K信号通路调控肾小球系膜细胞增殖。
목적：탐토1,25-이간기유생소D3[1,25(OH)2D3]대인신소구계막세포증식적영향급기재신소구계막세포조공중대mTOR/p70s6K신호통로적작용。방법체외배양인신소구계막세포，취전대배양지3~7대세포분위4조：정상대조조（가함5%태우혈청DMEM배양기），VD조[가1,25(OH)2D310-8 mol/L]，R조（가뢰파매소5 mg/L），R+VD조[가뢰파매소5 mg/L급1,25(OH)2D310-8 mol/L]，간예48 h。채용세포증식여활성검측시제합CCK-8검측세포증식정황，류식세포술검측세포주기시상분포，면역형광법검측세포중mTOR、p70s6K적표체정황。결과정상대조조、VD조、R조、R+VD조：（1）흡광도치（A450）의차강저，조간다중비교차이균유통계학의의（균P<0.05）；억제솔（IR）의차승고。（2）G1기세포비례의차증가，S기、G2/M기의차감소，증식지수（PI）의차강저，제R조여VD조비교차이무통계학의의외，기여조간다중비교차이균유통계학의의。（3）계막세포중mTOR、p70s6K단백표체강도균의차강저，제R조여VD조비교차이무통계학의의외，기여조간다중비교차이균유통계학의의。결론1,25(OH)2D3가현저억제인계막세포증식，차기가능통과억제mTOR/p70s6K신호통로조공신소구계막세포증식。
Objective To investigate the effects of 1,25-dihydroxyvitamin D3[1,25 (OH)2D3] on cell proliferation in hu?man glomerular mesangial cells and it′s effects on the regulation of mTOR/p70s6K signaling pathway in this cell line. Meth?ods The cultured human mesangial cells at passage 3-7 were divided into four groups:control group,VD group (addition of 10-8 mol/L of 1,25-dihydroxyvitamin D3 ),R group (addition of 5 mg/L of rapamycin) and R+VD group(addition of 5 mg/L ra?pamycin combined with 10-8 mol/L of 1,25-dihydroxyvitamin D3). Drug incubation last 48 h. The effect of mesangial cell pro?liferation was measured by CCK-8 colorimetric assay. The cell cycles were measured by flow cytometry. The expression of mTOR and p70s6K were detected by immunofluorescence. Results (1) The absorbance of A450 was higher in control group than that in VD group than that in R group than that in R+VD group. But the inhibition rate (IR) was lower in control group than that in VD group than that in R group than that in R+VD group. All comparisons were of statistic significance. ( 2) Cells in G1 phase were higher while cells in G2/M and S phases as well as proliferation rate (PI) were lower in control group than those in VD group than those in R group than those in R+VD group. All comparisons were of statistic significance except in?dexes between group R and group VD. (3) mTOR and p30s6K expressions in mesangial cells were higher in control group than those in VD group than those in R group than those in R+VD group. All comparisons were of statistic significance ex?cept indexes between group R and group VD. Conclusion 1,25-dihydroxyvitamin D3 might inhibit mesangial cell prolifera?tion significantly through mTOR/p70s6K signaling pathways.