“调神畅志”法针刺对帕金森伴便秘模型大鼠结肠组织PI3K/AKT/eNOS信号通路的影响

Effect of‘Tiaoshen Changzhi’Acupuncture on PI3K/AKT/eNOS Signaling Pathway in Colon Tissues of Model Rats with PD Constipation

目的:探讨“调神畅志”针法对帕金森(PD)伴便秘大鼠结肠组织磷脂酰肌醇3-激酶/蛋白激酶B/内皮型一氧化氮合酶(PI3K/AKT/eNOS)信号通路关键标志物(PI3K、AKT、p-Akt及eNOS)的影响,并探究此针法治疗的机制。方法:采用颈背部皮下注射鱼藤酮的方法制造PD伴便秘大鼠模型,并用APO诱导检测。40只造模成功大鼠随机分为模型组、西药组、常规针刺组与调神畅志组,每组10只,另空白组、假手术组各10只,共计60只,予相应干预4周。采用Western blot法检测各组大鼠远端结肠组织中PI3K、AKT、p-Akt蛋白表达和eNOS含量。结果:与空白组相比,模型组大鼠远端结肠组织中PI3K、AKT及p-Akt蛋白表达显著降低,eNOS含量显著升高,差异具有统计学意义(P<0.01);与模型组比较,西药组、常规针刺组和调神畅志组远端结肠组织PI3K、AKT及p-Akt蛋白表达均明显升高,eNOS含量均明显降低,差异具有统计学意义(P<0.05或P<0.01);与常规针刺组比较,调神畅志组PI3K、AKT及p-Akt蛋白表达明显较高,eNOS含量显著较低,差异具有统计学意义(P<0.05)。结论:针刺能够改善PD伴便秘大鼠模型便秘症状,且调神畅志针刺优于常规针刺,PI3K/AKT/eNOS信号通路的激活是调神畅志针法的治疗机制之一,其作用机制是通过上调大鼠远端结肠组织中PI3K/AKT/eNOS信号通路中PI3K、AKT、p-Akt的蛋白表达和降低eNOS含量来实现的。

Objective:To investigate the effect of Tiaoshen Changzhi acupuncture on the key proteins of PI3K,AKT,p-Akt and eNOS of PI3K/AKT/eNOS signaling pathway in the colon of rats with Parkinson’s disease(PD)constipation,and to explore the action mechanism of the therapy.Methods:PD constipation rat model was made by subcutaneous injection of rotenone into the back of neck,and detected by APO.40 successful model rats were randomly divided into the model group,the Western medication group,the routine acupuncture group and the Tiaoshen Changzhi acupuncture group,with 10 rats in each group.In addition,there were 10 rats in the blank group and 10 rats in the sham operation group,respectively.A total of 60 rats were given corresponding intervention for 4 weeks.Western blot was adopted to detect the protein expressions of PI3K,AKT,p-Akt and eNOS in distal colon tissues.Results:Compared with those in the blank group,the protein expressions of PI3K,AKT and p-Akt in the distal colon were significantly decreased,and the content of eNOS was significantly increased in the model group(P<0.01).Compared with those in the model group,the protein expressions of PI3K,AKT and p-Akt in distal colon tissues were increased significantly,and the content of eNOS was significantly decreased in the Western medication group,the routine acupuncture group and the Tiaoshen Changzhi acupuncture group(P<0.05,P<0.01).Compared with those in the routine acupuncture group,the protein expressions of PI3K,AKT and p-Akt were significantly increased,and the content of eNOS was significantly decreased in the Tiaoshen Changzhi acupuncture group(P<0.05).Conclusion:Acupuncture can relieve PD constipation of the model rats,and the therapeutic effect of Tiaoshen Changzhi acupuncture is better than that of routine acupuncture.Its therapeutic mechanism may be achieved by up-regulating the protein expressions of PI3K,AKT and p-Akt and reducing the content of eNOS in PI3K/AKT/eNOS signaling pathway in distal colon tissues.