α1肾上腺素受体在缺氧性肺动脉平滑肌细胞增殖中的作用 .pdfVIP

α1肾上腺素受体在缺氧性肺动脉平滑肌细胞增殖中的作用

姚香兰;薛全福;赵琪平

【摘要】目的:探讨肾上腺素α1受体(α1AR)及β2受体(β2AR)在缺氧性肺动脉平

滑肌细胞(PASMCs)增殖中的作用及分子机制.方法:用贴块法培养新生牛肺动脉平

滑肌细胞至第6代,并作免疫组化染色鉴定.细胞在6.6%O2环境中常压缺氧6、

12、24h,用[3H]-TdR掺入法反映DNA合成测定PASMCs的增殖,用Fura-

2/AM测定胞内Ca2+浓度.用Northernblotting法观察PASMCs的c-fos、c-

myc基因表达和α1ARmRNA、β2ARmRNA的变化.加用不同的受体激动剂和抑

制剂,探讨α1AR被激活和抑制后,以及β2AR激活后对上述变化的影响.结果:单纯

缺氧,PASMCsDNA合成即明显增加,α1AR激活后,增加更明显;而α1AR抑制后,明

显降低(P0.01).β2AR激活时无明显变化.缺氧后胞内游离钙浓度也增高

(P0.01),PASMCs上c-fos和c-myc基因表达增强(P0.05),α1ARmRNA和

β2ARmRNA水平均显著增高(P0.01).结论:结果提示缺氧刺激PASMCs,使胞内

钙浓度增加,c-fos和c-myc基因表达增强,引起PASMCs增殖,这一增殖是通过

α1AR介导的,但β2AR不起主要作用.缺氧时α1AR增多,既促进肺血管收缩,又加强

细胞增殖,在缺氧性肺血管收缩和肺血管重建导致的肺动脉高压中起一定的作

用.%AIM:Toinvestigatetheroleofα1andβ2adrenoceptors(α1ARand

β2AR)intheproliferationofhypoxicpulmonaryarterysmoothmuscle

cells(PASMCs).METHODS:PASMCswereisolatedbyanexplantmethod

fromneonatalbovinepulmonaryarteries.TheculturedPASMCswere

exposedto6.6％O2for6h,12hand24h.Themethodof[3H]-TdR

incorporationwasusedtomeasuretheproliferationofPASMCs.[Ca2+]i

wasassayedwithFura-2/AM.ThemRNAexpressionofα1AR,β2AR,c-fos

andc-mycwasdeterminedbyNorthernblotting.Theeffectsofactivation

ofα1ARandβ2AR,andinhibitionofα1ARontheaboveindexeswere

observedbytreatingPASMCswithdifferentARagonistsandantagonists

underhypoxiccondition.RESULTS:SignificantincreaseinTdRincorporation

inhypoxicPASMCswithα1ARactivationwasobserved,andmarked

decreaseinthatwasinducedbyα1ARinhibition.However,nosignificant

changewasfoundafterβ2ARactivation.[Ca2+]i,themRNAexpressionof

c-fos,c-myc,α1ARandβ2ARinPASMCswereincreasedafter

hypoxia.CONCLUSION:Hypoxiainducestheincreasein[Ca2+]iandmRNA

expressionofc-fosandc-myc,leadingtotheproliferationof

PASMCs.Theh