To Explore The Mechanism Of Action Of Herbal Cake-partitioned Moxibustion In Regulating MMP-9 And Collagen Fibers To Stabilize Vulnerable AS Plaques Based On The RhoA/ROCK1 Signaling Pathwa

Objective: Observing the effect of the herbal-cake-separated moxibustion on the expressions of RhoA 、 ROCK1 、 MMP-9 and the content of collagen fibers in rabbits with vulnerable atherosclerotic plaques to explore the possible mechanism of herbal-cake-separated moxibustion to stabilize AS vulnerable plaques.Methods: 70 New Zealand white rabbits weighing 2±0.2kg were randomly divided into a control group(10 rabbits)and an operation group(60 rabbits)after 1 week of adaptive feeding.The high-fat diet + balloon injury + gene transfection +drug-induced method was applied to rabbits in the operation group to prepare a rabbit model of vulnerable atherosclerotic plaque.After abdominal arterial balloon injury,the rabbits in the operation group were randomly divided into a model group,a direct moxibustion group,a herbalcake-separated moxibustion group,an atorvastatin group,and an inhibitor group.After grouping,each group received corresponding interventions for 8weeks.Direct moxibustion and herbal-cake-separated moxibustion were implemented in rabbits of the direct moxibustion group and the herbal-cakeseparated moxibustion group.The rabbits in the atorvastatin group were given atorvastatin.The rabbits in the inhibitor group were given fasudil hydrochloride via the ear vein injection.The rabbits of the control group and the model group were only bound without any interventions.After the treatment,all rabbits except the control group were transfected with an Ad5-p53 recombinant vector for two weeks.Then viper venom and histamine were applied to rabbits 24 and 48 hours before sacrifice to generate AS vulnerable plaques.ELISA method was used to detect the serum TC,TG,HDL-C and LDL-C content,HE staining was used to observe the morphology of the intima and media of the rabbit abdominal aorta,the immunohistochemistry was applied to detect the content of RhoA,ROCK1 and MMP-9 proteins expression in the abdominal aorta,and the content of collagen fibers was determined by Sirius scarlet staining.Results:1.Serum lipid: Compared with the control group,the HDL-C content in the model group was significantly reduced(P<0.01),while the TC,TG,and LDL-C content was significantly increased(P<0.01);compared with the model group,the content of HDL-C in the direct moxibustion group,the herbal-cake-separated moxibustion group and the atorvastatin group were significantly increased(P<0.01,P<0.05);compared with the model group,the TC,TG,LDL-C content of the direct moxibustion group,the herbal-cakeseparated moxibustion group and the atorvastatin group were significantly shortened(P<0.01,P<0.05).2.Abdominal aorta morphology: HE staining showed that the rabbit abdominal aorta in the model group was structurally disordered and the intima was thickened;after the intervention,the plaque cell morphology in each treatment group was more complete than the model group,and the intima-media boundary was greater obvious.3.The expression of RhoA,ROCK1,and MMP-9 in the abdominal aorta:Compared with the control group,the expression of RhoA,ROCK1 and MMP-9 of rabbits in the model group was significantly increased(P<0.01);compared with the model group,the expressions of RhoA and ROCK1 in the direct moxibustion group,the herbal-cake-separated moxibustion group,the atorvastatin group,and the inhibitor group all decreased significantly(P<0.01);compared with the model group,the expression of MMP-9 in the herbal-cake-separated moxibustion group,the atorvastatin group and the inhibitor group reduced significantly(P<0.01).4.Collagen content in the abdominal aorta: Compared with the control group,the content of collagen fibers in rabbits of the model group was significantly reduced(P<0.01);compared with the model group,the content of collagen fibers in the direct moxibustion group,the herbal-cake-separated moxibustion group,and the atorvastatin group increased significantly(P<0.01,P<0.05),the content of collagen fibers in the inhibitor group showed an upward trend but no significant difference(P>0.05).Conclusion:1.The herbal-cake-separated moxibustion can effectively regulate the blood lipid level of rabbits with vulnerable atherosclerosis plaques.2.The mechanism of herbal-cake-separated moxibustion in stabilizing vulnerable atherosclerosis plaques may be related to inhibiting the expressions of RhoA/ROCK1 and MMP-9,and increasing the content of collagen fibers.