CAJ | 학술논문

目的：研究胞内环磷酸腺苷（cAMP）水平提高对小鼠心肌成纤维细胞（MCFs）表达结缔组织生长因子（CT‐GF）的影响及其信号通路。方法采用出生1周内C57BL乳鼠心脏分离培养原代MCFs，第3代MCFs使用腺苷酸环化酶激活剂Forskolin干预培养后检测细胞CTGF、蛋白激酶A（PKA）、p44/42丝裂素活化蛋白激酶（MAPK）及磷酸化p44/42MAPK表达情况。分别给予磷酸化p44/42MAPK抑制剂PD98509及PKA抑制剂Rp‐cAMPS阻断相关信号节点，检测细胞内PKA、p44/42MAPK、磷酸化p44/42MAPK以及CTGF表达变化。结果Forskolin干预培养MCFs细胞后，胞内cAMP水平提高，细胞活力下降，PKA表达上升，p44/42MAPK总蛋白无明显改变，p44/42MAPK磷酸化水平下降，CTGFmRNA及蛋白表达下降；给予PD98509抑制p44/42MAPK磷酸化后，PKA表达上升，CTGFmRNA及蛋白表达下调；Rp‐cAMPS抑制PKA活化后，p44/42MAPK磷酸化水平升高，CTGFmRNA及蛋白表达上调。结论Forskolin可以通过提高MCFs胞内cAMP水平抑制CTGF的表达，其作用信号通路为高水平cAMP上调PKA表达，降低下游p44/42MAPK磷酸化水平，抑制CTGF的表达。
목적：연구포내배린산선감（cAMP）수평제고대소서심기성섬유세포（MCFs）표체결체조직생장인자（CT‐GF）적영향급기신호통로。방법채용출생1주내C57BL유서심장분리배양원대MCFs，제3대MCFs사용선감산배화매격활제Forskolin간예배양후검측세포CTGF、단백격매A（PKA）、p44/42사렬소활화단백격매（MAPK）급린산화p44/42MAPK표체정황。분별급여린산화p44/42MAPK억제제PD98509급PKA억제제Rp‐cAMPS조단상관신호절점，검측세포내PKA、p44/42MAPK、린산화p44/42MAPK이급CTGF표체변화。결과Forskolin간예배양MCFs세포후，포내cAMP수평제고，세포활력하강，PKA표체상승，p44/42MAPK총단백무명현개변，p44/42MAPK린산화수평하강，CTGFmRNA급단백표체하강；급여PD98509억제p44/42MAPK린산화후，PKA표체상승，CTGFmRNA급단백표체하조；Rp‐cAMPS억제PKA활화후，p44/42MAPK린산화수평승고，CTGFmRNA급단백표체상조。결론Forskolin가이통과제고MCFs포내cAMP수평억제CTGF적표체，기작용신호통로위고수평cAMP상조PKA표체，강저하유p44/42MAPK린산화수평，억제CTGF적표체。
Objective To examine the effects of the increased levels of cyclic adenosine monophosphate (cAMP)on the ex‐pression of connective tissue growth factor(CTGF)in mouse cardiac fibroblasts(MCFs)and the related signal transduction path‐way.Methods MCFs from the hearts of 1‐week‐old C57BL mice were isolated and primarily cultured.The expression levels of CTGF ,protein kinase A(PKA) ,p44/42 mitogen activated protein kinase(MAPK)and phospho‐p44/42 MAPK were detected in 3‐generation MCFs treated with Forskolin(an activating agent of cAMP) ,PD98509(an inhibitor of phospho‐p44/42 MAPK)or Rp‐cAMPS(an inhibitor of PKA).Results The levels of cAMP were increased ,the cell viability decreased ,the expression levels of PKA and CTGF increased and the expression levels of phospho‐p44/42 MAPK declined in MCFs treated with Forskolin. The PKA expression levels were increased while the expression levels of CTGF decreased in MCFs treated with PD 98509.Rp‐cAMPS could inhibit the expression of PKA ,and upregulate the expressions of phospho‐p44/42 MAPK and CTGF.Conclusion Forskolin can inhibit the CTGF expression levels by increasing the intracellular level of cAMP.The involved mechanism is that high levels of cAMP upregulate the PKA expression level ,reduce the phosphorylation level of the downstream target p44/42 MAPK and inhibit the CTGF expression.