丹参通络解毒汤联合内皮祖细胞对心肌缺血再灌注损伤大鼠PI3K/Akt信号通路的影响

Effects of Danshen Tongluo Jiedu Decoction Combined with Endothelial Progenitor Cells on PI3K/Akt Signaling Pathway of MIRI Rats

目的观察丹参通络解毒汤联合内皮祖细胞(EPCs)对心肌缺血再灌注损伤(MIRI)大鼠PI3K/Akt信号通路的影响,探讨其可能的作用机制。方法密度梯度离心法及差数贴壁法分离EPCs,SD大鼠随机分为假手术组、模型组、EPCs组、丹参通络解毒汤组、丹参通络解毒汤+EPCs组(联合组)、丹参通络解毒汤+EPCs+PI3K阻断剂组(阻断剂组),每组10只。分别给予相应处理,14 d后生物信号采集系统测定大鼠心功能,HE染色观察大鼠心肌组织病理变化,Western blot检测大鼠心肌组织p-PI3K、p-Akt蛋白表达。结果与假手术组比较,模型组大鼠心肌组织损伤明显,心肌纤维排列紊乱、肿胀,炎性细胞大量浸润,部分心肌细胞坏死;左心室形成压(LVDP)、左心室收缩压(LVSP)、左心室射血分数(LVEF)明显降低(P<0.01),p-PI3K、p-Akt蛋白表达明显升高(P<0.01)。与模型组比较,EPCs组、丹参通络解毒汤组及联合组心肌组织损伤程度均有不同程度改善;LVDP、LVSP、LVEF明显升高,p-PI3K、p-Akt蛋白表达明显升高,差异均有统计学意义(P<0.01),且联合组效果优于EPCs组和丹参通络解毒汤组(P<0.01)。与EPCs组、丹参通络解毒汤组、联合组比较,阻断剂组心肌组织损伤程度加重,LVDP、LVSP、LVEF明显降低,p-PI3K、p-Akt蛋白表达明显下降,差异均有统计学意义(P<0.01)。结论丹参通络解毒汤联合EPCs移植可保护大鼠MIRI,其机制可能与调节PI3K/Akt信号通路有关。

Objective To observe the effects of Danshen Tongluo Jiedu Decoction combined with endothelial progenitor cells(EPCs)on PI3K/Akt signaling pathway of rats with myocardial ischemia reperfusion injury(MIRI);To discuss its possible mechanism.Methods The density gradient centrifugation and differential adhesion method were adopted for EPCs isolation and culture.SD rats were randomly divided into sham-operation group,model group,EPCs group,Danshen Tongluo Jiedu Decoction group,Danshen Tongluo Jiedu Decoction+EPCs group(combination group),Danshen Tongluo Jiedu Decoction+EPCs+PI3K blocker group(blocker group),with 10 rats in each group and given corresponding treatment respectively.14 days later,the cardiac function was determined by bioinformatics collection system;pathological changes of the myocardial tissue were examined by HE staining method;Western blot was used to detect the protein expressions of p-PI3K and p-Akt.Results Compared with sham-operation group,rats in the model group had obvious myocardial tissue damage,disordered myocardial fiber arrangement,swelling,massive infiltration of inflammatory cells,necrosis of some myocardial cells;left ventricular developed pressure(LVDP),left ventricular systolic pressure(LVSP),and left ventricular ejection fractions(LVEF)decreased significantly;the expressions of p-PI3K and p-Akt protein significantly increased(P<0.01).Compared with the model group,the degree of myocardial tissue injury in the EPCs group,Danshen Tongluo Jiedu Decoction group and combination group was improved to varying degrees;LVDP,LVSP,and LVEF significantly increased;the expressions of p-PI3K and p-Akt protein increased,with statistical significance(P<0.01);the effect of combination group was better than that of EPCs group and Danshen Tongluo Jiedu Decoction group(P<0.01).Compared with the EPCs group,Danshen Tongluo Jiedu Decoction group and combination group,the degree of myocardial tissue damage in the blocker group was aggravated;LVDP,LVSP,and LVEF were significantly reduced,and the expressions of p-PI3K and p-Akt decreased,with statistical significance(P<0.01).Conclusion Danshen Tongluo Jiedu Decoction combined with EPCs transplantation can protect rats with MIRI,the mechanism could be related to regulating PI3K/Akt signaling pathway.