肺动脉内皮和平滑肌对缺氧损伤反应性的差异优选课件.ppt

1、 China 2020/10/181研究目的比较缺氧情况下肺动脉内皮细胞(Pulmonary arterial endothelial cells,PAECs)和平滑肌细胞(smooth muscle cells,PASMCs)在增殖活性、活性物质分泌方面的差异。2020/10/182方法细胞培养：胎牛PAECs和PASMCs取自同一肺小动脉段，在 含20胎牛血清的RPMI-1640培养基中于37 5%CO2 孵箱中培养。细胞增殖活性：采用氚标胸腺嘧啶核(3H-thymidine,3H-TdR)掺人和流式细胞技术测定。放免测定：细胞cGMP(用用于评估细胞内NO 的产生)、ET-1、ATII、

2、ANP等。斑点杂交：测定ET-1 mRNA 表达 2020/10/183结果与讨论一、PAECs与PASMCs增殖活性的比较二、缺氧时细胞内cGMP的变化三、缺氧时ET1分泌和表达改变四、缺氧时ATII变化的比较五、缺氧时细胞ANP分泌的变化六、缺氧时细胞EDLF分泌的变化2020/10/184一、PAECs与PASMCs增殖活性的比较3H-TdR掺人的时间掺入关系Time response of3H-TdrincorporationTime elaped(Hour)3H-Tdr incorporation(dpm/104 cells)04812162001000020000300004000

3、0PAEC r=0.983564PASM r=0.9964772020/10/1853H-Tdr incorporation of PASMs andPAECs cultured under hypoxiaOxygen concentration3H-Tdr incorporation(dpm/104 cells)21%2.5%0.5%0100002000030000PASMPAEC2020/10/186Flowcytometry data100%21%0.5%21%0.5%020406080100G1SG2/MPAECPASM2020/10/187PAECcGMP Content(pmol/

4、106cells)-LNA+LNA04812162021%O20.5%O2PASMcGMP Content(pmol/106cells)-LNA+LNA04812162021%O20.5%O22020/10/188ET-1 SecretionTime(h)ET-1(pg/ml)01224364801020304021%0.5%ET-1 mRNATime(h)IOD(100%of control)012243648010020030021%0.5%缺氧时内皮细胞ET-1分泌和表达的改变2020/10/189ET-1 SecretionTime(h)ET-1(pg/104 cells)012243

5、64805001000150021%0.5*ET-1 mRNATime(h)IOD(100%of control)012243648030609012021%0.5%2020/10/1810Relationship of ET-1 and Cell ProliferationGroups3H-Tdr incorporation(dpm/104 cells)-BQ123+BQ1230300060009000120001500021%O20.5%O22020/10/1811ACE ActivityTime(h)ACE activity(nmol/min/105 cells)012243648090

6、18027036021%O22.5%O20.5%O2*缺氧时内皮细胞ACE分泌增加2020/10/1812ATII secretin by PAECTime(h)ATII content(pg/105 cells)01224364805010015021%O22.5%O20.5%O22020/10/1813Effect of LNA on ATII secretionTime(h)ATII secretion(pg/105 cells)122405010015021%0.5%21%+L0.5%+L2020/10/18142020/10/1815ATII secretin by PASMTime

7、(h)ATII content(pg/105 cells)012243648010020030021%O22.5%O20.5%O2P0.001 vs 21%O22020/10/18162020/10/1817Relationship of ATII and Cell ProliferationGroups3H-Tdr incorporation(dpm/104 cells)-Captopril+Captopril0300060009000120001500021%O20.5%O2*#2020/10/1818ANPsecretionGroupANP(pg/105 cells/24h)PAECPA

8、SM025507521%O20.5%O2ANP contentGroup relative ANP content(IOD)PAECPASM01020304021%O20.5%O22020/10/1819EDLF secretionGroupEDLF(pg/105 cells/24h)PAECPASM010020030040021%O20.5%O22020/10/1820缩血管物质舒血管物质ET-1 ET-1 ETETmRNA mRNA ACE ACE ATII ATII EDLF EDLF ANP ANP NO NO 内皮细胞平滑肌细胞无缩血管物质：内皮细胞分泌增加，平缩血管物质：内皮细胞分泌增加，平滑肌细胞分泌减少滑肌细胞分泌减少舒血管物质：内皮细胞分泌减少，平舒血管物质：内皮细胞分泌减少，平滑肌细胞分泌增加滑肌细胞分泌增加结论：结论：缺氧使血管活性物质分泌失衡，可能缺氧使血管活性物质分泌失衡，可能是肺血管收缩的重要机制是肺血管收缩的重要机制失衡与制约失衡与制约小结与结论2020/10/1821谢谢您的聆听与观看THANK YOU FOR YOUR GUIDANCE.感谢阅读！为了方便学习和使用，本文档的内容可以在下载后随意修改，调整和打印。欢迎下载！汇报人：XXX日期：20XX年XX月XX日