隔药饼灸对动脉粥样硬化易损斑块兔RhoA/ROCK1信号通路及PI3K、Akt、mTOR表达的影响

Effects of Herbal-cake-separated Moxibustion on Expressions of RhoA/ROCK1 Signaling Pathway and PI3K,Akt,mTOR in Rabbits with Atherosclerotic Vulnerable Plaques

目的观察隔药饼灸对动脉粥样硬化(AS)易损斑块兔腹主动脉RhoA/ROCK1信号通路及PI3K、Akt、mTOR蛋白表达的影响,探讨其稳定AS易损斑块的作用机制。方法70只雄性新西兰兔随机分为空白组、模型组、隔药饼灸组、直接灸组、西药组和抑制剂组。采用高脂饲料喂养+腹主动脉球囊损伤+基因转染+药物触发斑块破裂制备AS易损斑块模型。各组分别给予相应干预,连续8周。采用ELISA检测血清载脂蛋白A(APO-A)、载脂蛋白B(APO-B)含量,HE染色观察腹主动脉内膜、中膜增生情况并测量内膜、内膜-中膜厚度,免疫组化染色检测Ras同源物基因组成员A(RhoA)、Rho同源物相关卷曲螺旋蛋白激酶1(ROCK1)蛋白表达,Western blot检测腹主动脉磷脂酰肌醇3激酶(PI3K)、蛋白激酶B(Akt)、哺乳动物雷帕霉素靶蛋白(mTOR)蛋白表达。结果与空白组比较,模型组兔血清APO-A含量显著减少(P<0.01),APO-B含量显著增加(P<0.01),内膜、内膜-中膜厚度增加,腹主动脉RhoA、ROCK1、PI3K、Akt、mTOR蛋白表达显著升高(P<0.01);与模型组比较,隔药饼灸组兔血清APO-A含量显著增加(P<0.01),APO-B含量显著减少(P<0.01),内膜、内膜-中膜厚度减少,腹主动脉RhoA、ROCK1、Akt、mTOR蛋白表达显著降低(P<0.01,P<0.05);与直接灸组比较,隔药饼灸组兔血清APO-B含量显著减少(P<0.01);与隔药饼灸组比较,抑制剂组兔腹主动脉PI3K蛋白表达显著降低(P<0.05)。结论隔药饼灸可能通过抑制RhoA/ROCK1信号通路,降低Akt、mTOR蛋白表达,促进细胞自噬,稳定AS易损斑块。

Objective To observe the effects of herbal-cake-separated moxibustion on the expressions of RhoA/ROCK1 signaling pathway and PI3K,Akt,mTOR in rabbits with atherosclerotic vulnerable plaques;To explore its mechanism of stabilizing vulnerable plaques.Methods Totally 70 male New Zealand rabbits were randomly divided into blank group,model group,herbal-cake-separated moxibustion group,direct moxibustion group,Western medicine group and inhibitor group.Vulnerable plaques model was established by high-fat feeding+abdominal aortic balloon injury+gene transfection+triggered plaque rupture.Each group was given corresponding interventions for 8 weeks.ELISA was used to detect the contents of apolipoprotein A(APO-A)and apolipoprotein B(APO-B)in serum.HE staining was used to observe vascular intima and media hyperplasia and to measure vascular intima and intima-media thickness.Immunohistochemistry was used to detect the proteins expression of RhoA and ROCK1 in abdominal aorta,Western blot was used to detect the expressions of PI3K,Akt and mTOR protein.Results Compared with the blank group,the content of APO-A in serum of the rabbits in model group significantly reduced(P<0.01),the contents of APO-B significantly increased(P<0.01),the intima,intima-media thickness increased,and the expressions of RhoA,ROCK1,PI3K,Akt and mTOR protein in abdominal aorta increased significantly(P<0.01).Compared with the model group,the content of APO-A in serum of the rabbits in herbal-cakeseparated moxibustion group significantly increased(P<0.01),the content of APO-B significantly reduced(P<0.01),the intima and intima-media thickness decreased,and the expression of RhoA,ROCK1,Akt and mTOR protein in abdominal aorta significantly decreased(P<0.01,P<0.05).Compared with the direct moxibustion group,the content of APO-B in serum of the rabbits in herbal-cake-separated moxibustion group significantly reduced(P<0.01).Compared with the medicinal cake-separated moxibustion group,the PI3K protein expression in abdominal aorta of the rabbits in inhibitor group was significantly reduced(P<0.05).Conclusion Herbal-cake-separated moxibustion could decrease the expression of Akt and mTOR protein by inhibiting RhoA/ROCK1 signaling pathway,promote autophagy,and stabilize atherosclerosis vulnerable plaques.