中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
Chinese Journal of Experimental Surgery
2015年
11期
2779-2782
,共4页
陶海鹰%贺斌%卫爱林%陶凤华%李孝海%刘康
陶海鷹%賀斌%衛愛林%陶鳳華%李孝海%劉康
도해응%하빈%위애림%도봉화%리효해%류강
人参皂苷Rb1%雪旺细胞%增殖%细胞周期%增殖细胞核抗原
人參皂苷Rb1%雪旺細胞%增殖%細胞週期%增殖細胞覈抗原
인삼조감Rb1%설왕세포%증식%세포주기%증식세포핵항원
Ginsenoside Rb1%Schwann cell%Proliferation%Cell cycle%Proliferating cell nuclear antigen
目的 观察人参皂苷Rb1(GRb1)对体外培养大鼠雪旺细胞(SCs)增殖、细胞周期及增殖细胞核抗原(PCNA)表达的影响.方法 取3～5d无特定病原体(SPF)级SD大鼠20只,雌雄不限,体质量25～30 g,分离双侧坐骨神经行SCs体外培养,培养细胞经S-100免疫荧光染色标记鉴定.取对数生长期SCs加入GRb1培养24 h,调整GRb1的终质量浓度为10 mg/L(B组)、20 mg/L(C组)、50 mg/L(D组)、100 mg/L(E组)、500 mg/L(F组),对照组(A组)加入等量磷酸盐缓冲液(PBS).细胞增殖检测法[细胞计数试剂盒(CCK-8)法]检测细胞增殖活性,碘化丙锭(PI)染色流式细胞仪检测细胞周期,实时定量聚合酶链反应(Real-time PCR)及Western blot法检测SCs内PCNA基因与蛋白的表达.结果 SCs经S-100免疫荧光标记染色提示阳性细胞率达90％以上,CCK-8检测结果显示GRb1在10～ 500 mg/L浓度范围内可以促进SCs增殖,流式细胞仪检测结果显示GRb1在10 ～500 mg/L浓度范围内可以促进SCs细胞周期进程,Real-time PCR及Western blot检测结果显示GRb1处理组SCs内PCNA基因和蛋白表达均高于对照组(P＜0.05).结论 GRb1可以促进SCs增殖及细胞周期进程,并促进SCs内PCNA表达.
目的 觀察人參皂苷Rb1(GRb1)對體外培養大鼠雪旺細胞(SCs)增殖、細胞週期及增殖細胞覈抗原(PCNA)錶達的影響.方法 取3～5d無特定病原體(SPF)級SD大鼠20隻,雌雄不限,體質量25～30 g,分離雙側坐骨神經行SCs體外培養,培養細胞經S-100免疫熒光染色標記鑒定.取對數生長期SCs加入GRb1培養24 h,調整GRb1的終質量濃度為10 mg/L(B組)、20 mg/L(C組)、50 mg/L(D組)、100 mg/L(E組)、500 mg/L(F組),對照組(A組)加入等量燐痠鹽緩遲液(PBS).細胞增殖檢測法[細胞計數試劑盒(CCK-8)法]檢測細胞增殖活性,碘化丙錠(PI)染色流式細胞儀檢測細胞週期,實時定量聚閤酶鏈反應(Real-time PCR)及Western blot法檢測SCs內PCNA基因與蛋白的錶達.結果 SCs經S-100免疫熒光標記染色提示暘性細胞率達90％以上,CCK-8檢測結果顯示GRb1在10～ 500 mg/L濃度範圍內可以促進SCs增殖,流式細胞儀檢測結果顯示GRb1在10 ～500 mg/L濃度範圍內可以促進SCs細胞週期進程,Real-time PCR及Western blot檢測結果顯示GRb1處理組SCs內PCNA基因和蛋白錶達均高于對照組(P＜0.05).結論 GRb1可以促進SCs增殖及細胞週期進程,併促進SCs內PCNA錶達.
목적 관찰인삼조감Rb1(GRb1)대체외배양대서설왕세포(SCs)증식、세포주기급증식세포핵항원(PCNA)표체적영향.방법 취3～5d무특정병원체(SPF)급SD대서20지,자웅불한,체질량25～30 g,분리쌍측좌골신경행SCs체외배양,배양세포경S-100면역형광염색표기감정.취대수생장기SCs가입GRb1배양24 h,조정GRb1적종질량농도위10 mg/L(B조)、20 mg/L(C조)、50 mg/L(D조)、100 mg/L(E조)、500 mg/L(F조),대조조(A조)가입등량린산염완충액(PBS).세포증식검측법[세포계수시제합(CCK-8)법]검측세포증식활성,전화병정(PI)염색류식세포의검측세포주기,실시정량취합매련반응(Real-time PCR)급Western blot법검측SCs내PCNA기인여단백적표체.결과 SCs경S-100면역형광표기염색제시양성세포솔체90％이상,CCK-8검측결과현시GRb1재10～ 500 mg/L농도범위내가이촉진SCs증식,류식세포의검측결과현시GRb1재10 ～500 mg/L농도범위내가이촉진SCs세포주기진정,Real-time PCR급Western blot검측결과현시GRb1처리조SCs내PCNA기인화단백표체균고우대조조(P＜0.05).결론 GRb1가이촉진SCs증식급세포주기진정,병촉진SCs내PCNA표체.
Objective To investigate the effects of Ginsenoside Rb1 (GRb1) on proliferation, cell cycle and expression of proliferating cellular nuclear antigen (PCNA) in Schwann cells (SCs).Methods Twenty 3-5 days old specific pathogen free (SPF) SD rats were used, no matter male or female, weighing 25-30 g.The SCs were isolated from sciatic nerves of SD rats in vitro, and identified by S-100 immunofluorescence markers staining.SCs were divided into six groups: group A, GRb1 10 mg/L;group B, GRb1 20 mg/L, group C, GRb1 50 mg/L, group D, GRb1 100 mg/L, group E, GRb1 500 mg/L, and the control group treated by equal volume of phosphate buffer (PBS).Cell counting kit-8 (CCK-8) assay was used to detect the cell proliferation, cell cycle was analyzed by propidium iodide (PI) staining and flow cytometry (FCM) analysis, and mRNA and protein levels were detected by real-time quantitative polymerase chain reaction (Real-time PCR) and Western blotting, respectively.Results S-100 immunofluorescence results showed that more than 95％ cells were positive in culture.CCK-8 results showed that GRb1 10-500 mg/L could increase cell proliferation.PI staining results indicated that RBb1 could accelerate the cell cycle process.Real-time PCR and Western blotting results indicated GRb1 of 10-500 mg/L could increase PCNA mRNA and protein levels in SCs.Conclusion GRb1 could promote SCs proliferation, accelerate cell cycle process, and increase the expression of PCNA mRNA and protein.
