The Protective Effect Of Grain-sized Moxibustion On Spontaneously Hypertensive Rats And The Mechanism Of Rho/ROCK Vascular Endothelial Signaling Pathway In Aortic

Objective:Observe the therapeutic effect of grain-sized moxibustion on Spontaneously Hypertensive Rats(SHR) and explore the mechanism of grain-sized moxibustion on expression of anti vascular injury by regulating Rho/ROCK signaling pathway.Methods:SHRs were randomly divided into a model group and a moxibustion group. W KY rats were regard as a bland group and inspect the therapy effect of SHRS with mox ibustion.Blood pressure of every rats were measured before intervention and recordJust g rabed and fixed the rats of model group and bland group without any other interfere,1 times a day for 28 days.While grabed and fixed the rats of the moxibustion group to ge t moxibustion with seed-sized moxa cone at "zusanli" and "quchi" at the same time of e very day, each acupoint has 5 cones, take the moxa cone away when it left a third,1 ti mes a day for 28 days. After the last interfere, measured the blood pressure of all group s again. Take the blood and aorta of all animals with 10% chloral hydrate anesthesia. M easured used the colorimetric method to measured serum SOD, MDA, NO of all rats in order to observate resistant effect of oxidative stress of SHR by moxibustion. Taken t he enzyme linked immunosorbent assay method to test the serum IL-6, IL-10, hs-CRP,IC AM-1 of all rats so as to investigate resistant effect of in flammatory reaction of SHR b y moxibustion. Observed the effect of lipid-reduced with moxibustion by testing the seru m content of TG, CHOL, HDL, LDL, ox-LDL. Used the HE staining to watch the thora cic aorta morphological changes of all rats. Overlooked the different metabolic products between SHR and normal rats through metabonomics technology. The last one was inspe cted the expression of protein and gene related to Rho/ROCK signal pathway.Results:(1)Blood pressure:before the experiment,the blood pressure of model group is higher than the blank group (P<0.01), model group and moxibustion group have no difference (P> 0.05). Measured the Blood pressure after the experiment, the blood pressure of the blank group have no difference with before (P> 0.05).Compared with the blank group, the blood pressure of the model group is higher(P<0.01). Compared with the model group, moxibustion group is lower(P<0.01).(2)Oxidative stress related indicators:Compared with the blank group, the model group,the content of MDA is higher and the content of SOD is lower(P<0.01). Compared with the model group, the moxibustion group, MDA is lower and the content of SOD is higher(P<0.01).(3)NO content:Compared with the blank group, the content of NO of the model group is lower(P<0.01). Compared with the model group, the content of NO of the moxibustion group is higher(P<0.01)(4)Blood lipid:Compared with the blank group, the content of TG, CHOL, LDL and ox-LDL of model group is increased significantly, while HDL decreased significantly (P<0.01). There was no deference between the blank group and the moxibustion group. Compared with the model group, the content of TG, CHOL, LDL and ox-LDL of moxibustion group is decreased,while HDL increased(P<0.01).(5)Inflammatory factors:Compared with the blank group, the content of IL-6, ICAM-1 and hs-CRP of model group is increased significantly, while IL-10 decreased significantly(P<0.01).Compared with the model group,the content oflL-6, ICAM-1 and hs-CRP of moxibustion group is decreased, while HDL increased significantly(P<0.01).(6)The result of aortic morphology changes by HE:Compared with the model group, there is not obvious aortic vascular injury, just some endothelial cells lining phenomenon happened. The membrane elastic fiber is irregular, part of them break.(7)Through PCA, metabolomic of the blank group, model group and moxibustion group clustered well. There were 27 types of biological markers contributed to the difference between 3 groups,most of them realeted to amino acid metabolism, glucose metabolism and lipid metabolism.(8)Expression of Rho/ROCK signaling pathway related genes in aorta of rat by RT-PCR:Compared with the blank group, the gene expression of RhoA、ROCK1、VEGF、 VEGFR2、MLC of model group is increased significantly(P<0.01).Compared with the model group, the gene expression of RhoA、ROCK、VEGF. VEGFR2、MLC of moxibustion group is decreased significantly (P<0.01).(9)Expression of Rho/ROCK signaling pathway related proteins in aorta of rat by WB:Compared with the blank group, the protein expression of RhoA、ROCK1、MLC,VEGF、VEGFR2 of model group is increased up significantly(P<0.01). Compared with the model group, the protein expression of RhoA、ROCK1、MLC,VEGF、VEGFR2 of moxibustion group is decreased down significantly(P<0.01).Conclusion:(1) Grain-sized moxibustion can reduce blood pressure in spontaneously hypertensive rats, and has the function of anti oxidative stress, anti inflammation, reducing blood fat, protecting vascular endothelial function. (2)Metabonomics can be blank group, model group, moxibustion group rat grain successfully distinguished. Spontaneously hypertensive rats have abnormal metabolism, mainly in amino acid metabolism, glucose metabolism, abnormal lipid metabolism. (3) Grain-sized moxibustion can protect the vascular endothelium by inhibiting the Rho/ROCK signaling pathway in the aorta.