低強度脈沖超聲波對SD大鼠前軟骨干細胞增殖及細胞外基質(zhì)mRNA表達的影響
發(fā)布時間：2018-10-11 13:07
【摘要】：目的研究低強度脈沖超聲波對大鼠前軟骨干細胞增殖及細胞外基質(zhì)分泌的影響,并探討可能存在的機制。 方法分離新生大鼠乳鼠的前軟骨干細胞,利用免疫磁珠分選的方法分離純化大鼠前軟骨干細胞,利用細胞表面特異性標(biāo)記物FGFR-3進行免疫組化鑒定。取生長良好的第3代細胞,用胰酶消化,含血清培養(yǎng)基中和后重懸細胞,顯微鏡下計數(shù),以1.6X10~~3個/ml的密度接種于96孔板,分4個組(即空白對照組、5分鐘/天組、12分鐘/天組、20分鐘/天組),每組設(shè)4個復(fù)孔。置于37℃,飽和濕度,5%C02培養(yǎng)箱培養(yǎng)24h,待細胞貼壁后進行低強度脈沖超聲波干預(yù)(超聲波強度為100mW/cm~2,頻率為1MHz,脈沖間隔為20%)。刺激時將耦合劑均勻涂于超聲波探頭上,將探頭置于培養(yǎng)板底部進行刺激,刺激后繼續(xù)培養(yǎng)細胞24h。分別于刺激后第一天、第三天和第五天用CCK-8法檢測細胞增殖。另以2X10~5個/ml的密度接種細胞于直徑35mm的細胞培養(yǎng)皿,分組同前,每組設(shè)3個復(fù)孔。干預(yù)方法同前,刺激后間隔30分鐘提取總RNA,用RT-PCR法檢測Ⅱ型膠原、Aggrecan、TGF-β_1及Sox9基因的表達。 結(jié)果LIPU對SD大鼠PCSCs的增殖有促進作用,與對照組相比在第一天時只有20分鐘/天刺激組增殖比較明顯,差異有統(tǒng)計學(xué)意義;而在連續(xù)刺激三天和五天后各實驗組和對照組均有明顯的差異,但是各實驗組之間的差異并不顯著。LIPU對SD大鼠PCSCs的Ⅱ型膠原、Aggrecan、TGF-β_1及Sox9基因mRAN的表達均有明顯的促進作用,刺激天數(shù)越多mRAN的表達量越多,以20分鐘/天刺激組增高最為明顯。 結(jié)論LIPU可以促進SD大鼠PCSCs的增殖能力,并可以促進細胞外基質(zhì)mRAN的表達,這一效應(yīng)可能是通過TGF-β_1及Sox9基因所介導(dǎo)的。
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【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R329
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9A%E8%BF%87TGF-%CE%B2_1%E5%8F%8ASox9%E5%9F%BA%E5%9B%A0%E6%89%80%E4%BB%8B%E5%AF%BC%E7%9A%84.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R329
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