低强度脉冲超声波对SD大鼠前软骨干细胞增殖及细胞外基质mRNA表达的影响
发布时间：2018-10-11 13:07
【摘要】：目的研究低强度脉冲超声波对大鼠前软骨干细胞增殖及细胞外基质分泌的影响,并探讨可能存在的机制。 方法分离新生大鼠乳鼠的前软骨干细胞,利用免疫磁珠分选的方法分离纯化大鼠前软骨干细胞,利用细胞表面特异性标记物FGFR-3进行免疫组化鉴定。取生长良好的第3代细胞,用胰酶消化,含血清培养基中和后重悬细胞,显微镜下计数,以1.6X10~~3个/ml的密度接种于96孔板,分4个组(即空白对照组、5分钟/天组、12分钟/天组、20分钟/天组),每组设4个复孔。置于37℃,饱和湿度,5%C02培养箱培养24h,待细胞贴壁后进行低强度脉冲超声波干预(超声波强度为100mW/cm~2,频率为1MHz,脉冲间隔为20%)。刺激时将耦合剂均匀涂于超声波探头上,将探头置于培养板底部进行刺激,刺激后继续培养细胞24h。分别于刺激后第一天、第三天和第五天用CCK-8法检测细胞增殖。另以2X10~5个/ml的密度接种细胞于直径35mm的细胞培养皿,分组同前,每组设3个复孔。干预方法同前,刺激后间隔30分钟提取总RNA,用RT-PCR法检测Ⅱ型胶原、Aggrecan、TGF-β_1及Sox9基因的表达。 结果LIPU对SD大鼠PCSCs的增殖有促进作用,与对照组相比在第一天时只有20分钟/天刺激组增殖比较明显,差异有统计学意义;而在连续刺激三天和五天后各实验组和对照组均有明显的差异,但是各实验组之间的差异并不显著。LIPU对SD大鼠PCSCs的Ⅱ型胶原、Aggrecan、TGF-β_1及Sox9基因mRAN的表达均有明显的促进作用,刺激天数越多mRAN的表达量越多,以20分钟/天刺激组增高最为明显。 结论LIPU可以促进SD大鼠PCSCs的增殖能力,并可以促进细胞外基质mRAN的表达,这一效应可能是通过TGF-β_1及Sox9基因所介导的。
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【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R329
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【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R329
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