The Effects Of Zhilong Huoxue Tongyu Capsule And Its Components On Vascular Remodeling Of Hypertension

Objectives: To observe the effect of Zhilong Huoxue Tongyu Capsule and its components on the expression of MDA superoxide dismutase SOD content and blood vessels NF-κB-p65, ICAM-1 in rats, and to explore its action and mechanism in vascular remodeling of hypertension.Methods: The screening of male Wistar rats were 100, 12 were divided randomly into normal group, 76 were into surgery group and 12 were into sham group. The surgery group were ligated the left renal artery to build renal hypertensive rats. There were 77 successful rats after 2 weeks operatively to build hyperactivity type renal hypertensive rats. At the seventh week, the model successful rats were screened out of 60 randomly to divide into model group, captopril group, full formula group, buqi group,wentong jingluo group,huoxue huayu group. Each group had 10 rats. The rats were fed the corresponding drugs, once a day. The normal group, sham group and model group were given the same volume of physiological saline. After 4 weeks, the rats were took the blood of abdominal aortic to prepare serum. The chemical colorimetry was used to determinate serum MDA and SOD. Taking the thoracic aorta, fastened, entrapped with paraffin, section, HE staining, then observing the change of vascular middle membrane and measuring MT,LD,MT/LD. Immunohistochemistry was used to detect the expression of NF-κB-p65, ICAM-1 in the thoracic aorta. And further, western blot method was used to detect the expression of vascular ICAM-1.Results: 1. The impact of drugs on the rats blood pressure in renal hypertensive rats:(1) The basis blood pressure: There is no statistical difference among the each group.(2) The blood pressure after 2 weeks and 6 weeks of the surgery: the normal group and sham group are almost the same from a statistical point of view; compared with the sham group, the model group were obvious increase(P<0.05); there is no statistical difference among the each drug group.(3)The blood pressure after 4 weeks of treatment: the normal group and sham group are almost the same from a statistical point of view; Compared with the sham group, normal group and the each drug group, the model group was obvious increase(P<0.05);compared with the full formula group,the captopril group was obvious decrease(P<0.05); and compared with the huoxue huayu group, the buqi group was insignificant decrease(P>0.05); compared with the wentong jingluo group, the buqi group was obvious decrease(P<0.05).2. The impact of drugs on oxidative stress in renal hypertensive rats:(1)The impact of drugs on MDA in renal hypertensive rats :the normal group and sham group are almost the same from a statistical point of view; compared with the other groups was obvious decrease(P<0.05); compared with buqi group and huoxue huayu group, the model group was insignificant increase; compared with sham group, normal group and other drug group, the model group was obvious increase(P<0.05); compared with huoxue huayu group,the buqi group was insignificant increase(P>0.05). Compared with captopril group,the full formula group insignificant increase(P>0.05);compared with wentong jingluo group and huoxue huayu group, the full formula group obvious decrease(P<0.05);and compared with wentong jingluo group,the huoxue huayu group insignificant increase(P>0.05).(2)The impact of drugs on SOD in renal hypertensive rats :the normal group and sham group are almost the same from a statistical point of view; compared with the other groups, sham group and normal group obvious increase(P<0.05); compared with buqi group,wentong jingluo group and huoxue huayu group,the model group was insignificant decrease(P>0.05); compared with captopril group and full formula group,the model group was obvious decrease(P<0.05);compared with huoxue huayu group,the wentong jingluo group was insignificant decrease;compared with buqi group,the wentong jingluo group was insignificant increase; compared with full formula group and huoxue huayu group,the captopril group was insignificant increase;compared with huoxue huayu group(P>0.05),the full formula group was insignificant increase(P>0.05).3.The impact of drugs on change of rat thoracic aorta of renal hypertensive rats: Under the light microscope, smooth muscle cell in the middle membrane of normal group and sham group are arrange generally tidy, no hypertrophy and hyperplasia. Smooth muscle cell and elastic fibers in the middle membrane of model group were hyperplasia, increase in amount, disorganized, middle membrane were thickness, and elastic fibers were fracture, a little foam cells. After the treatment, compared with model group, the hyperplasia degree of middle membrane and elastic fibers of captopril group,full formula group and buqi group were obvious decrease; wentong jingluo and huoxue huayu group were bring down mildly. And compared with the full formula group and buqi group,captopril group was obvious decrease.4.The impact of drugs on MT,LD,MT/LD of thoracic aorta:(1)The impact of drugs on MT of thoracic aorta: There is no statistical difference between the sham group and normal group; compared with the other groups was obvious decrease(P<0.05);The impact of drugs on MT,LD,MT/LD of thoracic aorta :(1)The impact of drugs on MT of thoracic aorta : The normal group and sham group are almost the same from a statistical point of view; compared with the other groups was obvious decrease(P<0.05); compared with others drug group,model group was obvious increase(P<0.05);compare with full formula group,captopril group was insignificant decrease(P>0.05),compared with others drug group,captopril group was obvious decrease(P<0.05); compare with full formula group, buqi group was insignificant increase(P>0.05), compare with wentong jingluo group and huoxue huayu group, buqi group was insignificant decrease(P>0.05); compare with huoxue huayu group,wentong jingluo group insignificant decrease(P>0.05).(2)The impact of drugs on LD of thoracic aorta: The normal group and sham group are almost the same from a statistical point of view; compared with the other groups was obvious increase(P<0.05); compared with huoxue huayu group and wentong jingluo group, medol group was insignificant decrease(P>0.05),and compared with wentong jingluo group, huoxue huayu group was insignificant increase(P>0.05);compared with other drug group, model group was obvious decrease(P<0.05); compared with other drug group, captopril group was obvious increase; compared with full formula group, buqi group obvious decrease(P<0.05); compared with huoxue huayu group and wentong jingluo group insignificant increase(P>0.05).(3)The impact of drugs on MT/LD of thoracic aorta: the normal group and sham group are almost the same from a statistical point of view; compared with the other groups was obvious increase(P<0.05); compared with others drug group,model group was obvious decrease(P<0.05);compared with others drug group, captopril group was obvious increase(P<0.05); compare with full formula group and buqi group, captopril group was obvious decrease(P<0.05), compare with huoxue huayu group and wentong jingluo group, buqi group was obvious increase(P<0.05), compare with wentong jingluo group,huoxue huayu group was insignificant increase(P>0.05).5.The impact of drugs on the expression of thoracic aorta NF-κB(IHC): NF-κB expression was brown granules and mainly located in the thoracic aorta smooth muscle under the light microscope.NF-κB was a small amount of expression in the normal group and sham group;NF-κB was an amount of expression in the model group. After the treatment, NF-κB was an amount of expression in the model group, compared with model group, captopril group and full formula group were obvious decrease(P<0.05), buqi group, wentong jingluo group and huoxue huayu group were decrease. Compared with full formula group, model group was decrease, compared with huoxue huayu group, buqi group was obvious decrease, and there is no statistical difference between the buqi group and wentong jingluo group.6.NF-κB protein Expression in Rat aortic smooth muscle of IOD comparison in every group: the normal group and sham group are almost the same from a statistical point of view; compared with the other groups was obvious decrease(P<0.05); compared with others drug group,model group was obvious increase(P<0.05);compared with others drug group, captopril group obvious increase(P<0.05); compare with full formula group, wentong jingluo group was obvious increase(P<0.05); compare with huoxue huayu group,wentong jingluo group was obvious increase(P>0.05), and compare with buqi group,wentong jingluo group was insignificant decrease(P>0.05).7.The impact of drugs on the expression of thoracic aorta ICAM-1(IHC): ICAM-1 expression was brown granules and mainly located in the thoracic aorta smooth muscle under the light microscope. ICAM-1 was a small amount of expression in the normal group and sham group;ICAM-1 was an amount of expression in the model group. After the treatment, ICAM-1 was an amount of expression in the model group, compared with model group, captopril group and full formula group were obvious decrease(P<0.05), buqi group, wentong jingluo group and huoxue huayu group were decrease. Compared with full formula group, model group was decrease, compared with huoxue huayu group, buqi group was obvious decrease.8. ICAM-1 protein Expression in Rat aortic smooth muscle of IOD comparison in every group: the normal group and sham group are almost the same from a statistical point of view; compared with the other groups was obvious decrease(P<0.05); compared with others drug group,model group was obvious increase(P<0.05);compared with others drug group, captopril group obvious increase(P<0.05); compare with full formula group, wentong jingluo group was obvious increase(P<0.05); compare with huoxue huayu group,wentong jingluo group was obvious increase(P>0.05), and compare with buqi group,wentong jingluo group was insignificant decrease(P>0.05).9. ICAM-1 protein Expression in Rat aortic smooth muscle in every group by western blot: the normal group and sham group are almost the same from a statistical point of view; compared with the other groups was obvious decrease(P<0.05); compared with others drug group,model group was obvious increase(P<0.05);compared with others drug group, captopril group obvious increase(P<0.05); compare with full formula group, wentong jingluo group was obvious increase(P<0.05); compare with huoxue huayu group,wentong jingluo group was obvious increase(P>0.05), and compare with buqi group,wentong jingluo group was insignificant decrease(P>0.05).Conclusion: 1. Through partial ligating left renal artery, the model rats with renal hypertensive rats were preparated successfully. The effects of Zhilong Huoxue Tongyu Capsule and its components has obvious therapeutic effect on vascular remodeling in rats with renal hypertension. The mechanism of action are including oxidative stress, inflammation, vascular mechanisms. The different components of Zhilong Huoxue Tongyu Capsule in a different way to play a therapeutic effect on Vascular remodeling in Hypertensive Rats:(1)Buqi group has prominent role in lowering blood pressure.(2)Wentong jingluo group has prominent role in lowering MDA, NF-κB and ICAM-1 protein expression.(3)Huoxue huayu group has role in lowering NF-κB and ICAM-1 protein expression.After compatibility of the different components, it generates significant synergies and shows the superiority of TCM treatment of multiple targets.