| Objective: 1.Observe the effect of cake-separated moxibustion on the Rho A/Rock signaling pathway of AS rabbits,and explore the mechanism of this pathway in stabilizing AS fragile plaques.2.Observe the effect of cake-separated moxibustion on related protein expression and macrophages autophagy in the mediated pathway of Rho A/Rock-PI3K/Akt/m TOR,and explore the mechanism of anti-inflammatory and stabilizing plaque moxibustion.Methods: 70 healthy male New Zealand white rabbits weigh 1.8 to 2.2 kg.After one week of adaptive feeding,10 were randomly selected as the control group of the blank group and fed with ordinary feed.The remaining 60 were assigned to the surgical group and fed with high-fat feed.After two weeks of high-fat feeding,abdominal aortic balloon injury was performed on rabbits in the surgical group.5animals died after surgery,and 55 surviving rabbits were randomly divided into the model group(11),direct moxibustion group(11),cake-separated moxibustion group(11),inhibitor group(11),and western medicine group(11).The rabbits in the surgical group continued to feed high-fat feed for 8 weeks and intervened with corresponding interventions for a total of 8 weeks.After treatment,feed all animals with ordinary feed,and use Ad5-p53 recombinant vector to transfect all animals except the blank group for a total of 2 weeks on the first day after treatment.Chinese spots and viper venom(CRVV)and histamine were triggered twice for all animals except the blank group on the 13 th weekend(24 and 48 hours before the material).Use Elisa to detect 6 serum lipids: total cholesterol(TC),triacyl glycerin(TG),lowdensity lipoprotein cholesterol(LDL-C),HDL-C,apolipoprotein A,B(apo-A,apo-B);The ultrastructure and autophagy of the abdominal aorta were observed by transmission electron microscopy;the thickness of the endovascular and Intima-media wall thickness(IMT)was measured by HE staining;and the protein expression of the abdominal aorta Rho A,Rock1 and Beclin1 and changes in the LC3-II/I ratio were detected by Western Blot detection technology.Results:1.Blood lipids: Compared with the model group,The contents of TG,TC,and LDL in the direct moxibustion group,cake-separated moxibustion group,and western medicine group were significantly reduced(p<0.01,p<0.05),and the content of HDL and HDL/LDL ratio increased significantly(p<0.01,p<0.05).Compared with the direct moxibustion group,there was no significant change in the blood lipid level in the cake-separated moxibustion group and western drug group(p>0.05).Compared with the cake-separated moxibustion group,there was no significant change in the blood lipid in the western medicine group(p>0.05),and the APO-B content decreased significantly(p<0.05).The results showed that the three treatment groups had a significant curative effect on regulating the overall blood lipid level of AS fragile plaque rabbits,and the effects of the three groups are similar.It shows that cakeseparated moxibustion can effectively regulate blood lipid levels to achieve anti-AS spot stabilization.2.Apolipoprotein: Compared with the model group,the content of APO-A increased significantly(p<0.05)and the content of APO-B and APO-B/APO-A ratio decreased significantly(p<0.05,p<0.01)in the direct moxibustion group,cakeseparated moxibustion group,and western medicine group.Compared with the direct moxibustion group,the APO-B content decreased significantly(p<0.01),and there was no significant change in the content of APO-A and APO-B/APO-A ratio(p>0.05)in the cake-separated moxibustion group.There was no significant change in the average apolipoprotein in the western drug group(p>0.05).Compared with the cakeseparated moxibustion group,the APO-B content increased significantly(p<0.01),and there was no significant change in the content of APO-A and APO-B/APO-A ratio(p>0.05)in the western drug group.The results showed that the three treatment groups had a significant curative effect in regulating apolipoprotein content.In terms of adjusting the content of APO-B,the cake-separated moxibustion group has the best curative effect.In terms of adjusting the content of APO-A,the three treatment groups have the same effect.3.HE staining: Compared with the blank group,the endovascular and Intimamedia wall thickness of the model group thicken,and protruded into the tube cavity to form plaques,showing thrombosis.The membrane structure of the middle layer of the blood vessel is damaged,elastic fibers are irregularly arranged,the structure is not clear,and smooth muscle cells are cellularly loose,which shows a large number of foam cells.Compared with the model group,the degree of vascular injury in the direct moxibustion group is comparable to that in the model group.The degree of media damage in the cake-separated moxibustion group and the western medicine group was slightly reduced.No thrombosis was seen in the three treatment groups,and the thickness of the endovascular and Intima-media wall was significantly lower than that of the model group(p <0.05 or p <0.01).It shows that the treatment of cake-separated moxibustion can reduce the hyperplasia of endovascular and Intima-media walls,thus achieving anti-AS plaque stabilization.4.Ultra-micro transmission electron microscope inspection: Compared with the blank group,the blood vessel smooth muscle cells in the model group were transformed into foam cells,and a large number of fat droplets were distributed in foam pond species.A large number of collagen filaments were fractured and dissolved flocculently.The cell membrane and organelle were seriously damaged,and no autophagy was found.Compared with the model group,the damage degree of organelles in the direct moxibustion group,the cake-separated moxibustion group,and the inhibitor group are the same,and autophagy can be seen in the field of vision.the cell damage in the western drug group is the least,and some organelles and collagen fibers are retained,but there is no autophagy in the field of vision.It shows that the cake-separated moxibustion can promote the autophagy function of AS vulnerable plaque rabbit cells.5.WB test results:(1)Rho A/Rock protein expression: Compared with the model group,the inhibitor can reduce the expression of Rho A and Rock1 protein(p<0.05).There was no significant change in the expression of Rho A and Rock protein(p>0.05).The expression of Rho A and Rock1 protein decreased significantly in the Cakeseparated moxibustion group.The expression of Rho A protein decreased significantly(p<0.05)and there was no significant change in Rock1 in the western drug group.Compared with the direct moxibustion group.The expression of Rho A and Rock1 protein significantly decreased in the cake-separated moxibustion group(p<0.05).The expression of Rho A significantly decreased(p<0.05)and there was no significant change(p>0.05)in the expression of Rock in the western drug group.It shows that cake-separated moxibustion and western medicine can inhibit the expression of Rho A and Rock1 protein after the intervention,and the efficacy of the two is the same.(2)Autophagy-related protein expression: Compared with the blank group,the expression of Beclin1 protein increased significantly(p<0.01)and the ratio of LC3Ⅱand LC3Ⅱ/Ⅰ also tended to increase,but the difference was not significant(p>0.05).Compared with the model group,the protein expression of Beclin1,LC3Ⅱ in the direct moxibustion group was significantly increased(p<0.01),and the ratio of LC3Ⅱ/Ⅰ changed insignificantly(p>0.05);The protein expression of Beclin1,LC3Ⅱ,and LC3Ⅱ/Ⅰratio in the cake-separated moxibustion group was significantly increased(p<0.01,p<0.05);in the western drug group,Beclin1 protein expression had no significant change(p>0.05),and the ratio of LC3Ⅱ protein expression and LC3Ⅱ/Ⅰsignificantly increased(p<0.01);Beclin1,LC3Ⅱ protein expression and LC3Ⅱ/Ⅰratio in the inhibitor group significantly increased(p<0.01).Compared with the direct moxibustion group: the Beclin1,LC3Ⅱ protein expression and the LC3Ⅱ/I ratio in the cake-separated moxibustion group changed insignificantly(p>0.05);In the western drug group,Beclin1 protein expression was significantly decreased(p<0.01),LC3 Ⅱ protein expression and LC3 Ⅱ / Ⅰ ratio changed insignificantly(p>0.05).Compared with the cake-separated moxibustion group: Beclin1,LC3 II protein expression and LC3II/I ratio in the western drug group had no significant difference(p>0.05);Beclin1 protein expression in the inhibitor group was significantly increased(p<0.01),LC3 II protein expression and LC3Ⅱ/Ⅰ ratio had no significant difference(p>0.05).Compared with the western drug group,the Beclin1 protein expression in the inhibitor group was significantly increased(p<0.01),and LC3 Ⅱ protein expression and the LC3Ⅱ/Ⅰ ratio had no significant difference(p>0.05).It shows that after the intervention of medicated cake moxibustion,the autophagy level of cells increases.Conclusion:1.Cake-separated moxibustion has a good regulatory effect on the metabolism of blood lipids and apolipoproteins in rabbits with AS vulnerable plaques.2.Cake-separated moxibustion can improve the vascular intimal structure of rabbits with AS vulnerable plaques,inhibit intimal hyperplasia,and protect the vascular endothelium.3.The mechanism of anti-AS stable plaque of cake-separated moxibustion may mediate the low expression of the Rho A/Rock signaling pathway,thus promoting cell autophagy function. |
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