The Study On Mechanism Of Traditional Chinese Drug Of Supplementing Qi And Activating Blood Circulation Delay Vascular Aging Caused By Aging And Hypertension

As is known to all, population aging is one of the most important problems in modern society development, and cardiovascular disease is the leading causes of death of the old. Age is an independent risk factor of cardiovascular disease. Vascular aging is shown obviously in aging and cardiovascular disease; therefore, the research of vascular aging has become an hotspot in senile disease study. Vascular aging shows blood vessel wall thicken and harden on the levels of vascular endothelial cells, smooth muscle cells and extracellular matrix. Vascular aging exhibits intimal thicken and endothelial cell structure and function changes in vascular intima, which was discussed already in the preliminary study. Vascular smooth muscle cells are the main component of vascular media, which abnormal proliferation and migration is the main pathological changes of vascular aging. In this study, vascular aging is the research purpose, based on previous research, we chosen vascular smooth muscle cell as the breakthrough point to research vascular aging caused by aging and disease. In this study, vascular aging is the research purpose, based on previous research, we chosen vascular smooth muscle cell as the breakthrough point to research vascular aging caused by aging and disease. In this study, vascular aging caused by aging and cardiovascular disease is researched from the whole, cellular and molecular levels. To explore the mechanism and effort of traditional Chinese drug of supplementing qi and activating blood circulation delay aging, it is provide a scientific basis for delaying vascular aging.This study is divided into three parts:literature review, basic research and clinical research.1. Literature reviewBy sorting and summing up the literatures of aging, the traditional Chinese medicine and modern medicine theory of aging were summarized and the mechanisms of traditional Chinese medicine delay aging were elaborated. By reviewing the literature of arterial stiffness, the article of the common detection methods of arterial stiffness and its clinical significance and research progress were summarized.2. Basic research This part includs four basic experiments.Experiment I:Experimental study on the effect of aging and hypertension on aortic structure of ratsObjective:To observe the effect of aging and hypertension on arterial wall structure. Methods:The thoracic aorta of the aged and young rats, WKY and SHR rats were stained by HE staining and Masson staining. Results:HE staining results showed that the aortic of the old and SHR rats intima thicken, uplift, and tunica media thicken obviously, VSMC migrated to the intima and enlarged; Masson staining results showed that the relative content of aortic VSMC and collagen of the aged and SHR rats were significantly increased. Conclusion:Aging and high blood pressure leads to vascular structure variation, and the characteristics of aging on the blood vessels. It is suggested that high blood pressure could induce and promote vascular aging.Experiment?:Primary culture and identification of thoracic aorta vascular smooth muscle cell in ratsObjective:To investigate the primary culture method of rat thoracic aorta vascular smooth muscle cell (VSMC) and provide the important experimental material for the research of vascular pathological changes. Methods:The primary culture of rat thoracic aorta vascular smooth muscle cells were done by tissue-piece inoculation. The cultured cells were observed by phase contrast microscope for morphology and growth condition, and the cells were identified by immunocytochemical staining. Results:Under the microscope, the cultured cells showed typically "peak and valley" growth feature and expressed positive of smooth muscle a-actin by immunocytochemical staining. Conclusion:The method is simple and reliable, which could obtain highly purified VSMC with good structure and function and provide a good experimental material for cardiovascular disease research. Experiment?:The study on mechanism of traditional Chinese drug of supplementing qi and activating blood circulation delay vascular aging in aged ratsObjective:To observe the pathological changes of vascular aging in aged rats and the intervention effort and mechanism of extracts from Panax ginseng, Panax notoginseng(Burk.) and Ligusticum chuanxiong on vascular aging. Methods:The VSMC of aged and young rats were primarily cultured and divided into the young group, the old group, probucal group, low dose of Chinese medicine group and high dose of Chinese medicine group. The aging of cells were detected by SA-?-gal staining, the cell cycle was analyzed by flow cytometry analysis, the levels of MMP-9 and TGF?1 were detected by immunocytochemistry and the expression of MLCK was detected by immunofluorescence staining. The mRNA and protein expression of p16, cyclinD1, CDK4, pRb and PPARy were detected by RT-PCR and Western Bloting. Results:Compared with the young group, VSMC of the old group proliferated significantly, extracts from Panax ginseng, Panax notoginseng(Burk.) and Ligusticum chuanxiong could reduce the SA?-gal expression, arrest the cell cycle in G1 phase, reduce expression of MMP9, TGF?1 and increase the MLCK expression. The extract could increase the mRNA and protein expression of p16, Rb, PPARy while reducing cyclinDl and CDK4. Conclusion:The extracts from Panax ginseng, Panax notoginseng(Burk.) and Ligusticum chuanxiong can delay VSMC aging, which works by cytokines and p16-CyclinD/CDK-RB way to inhibit VSMC proliferation, migration, enhance the contraction of VSMC, reduce the vascular structure changes with aging and improve the elasticity of blood vessel.Experiment IV:The study on mechanism of traditional Chinese drug of supplementing qi and activating blood circulation delay vascular aging in SHR ratsObjective:To observe the pathological changes of vascular aging in SHR rats, and the effect and mechanism of extracts from Panax ginseng, Panax notoginseng(Burk.) and Ligusticum chuanxiong on vascular aging. Methods:The VSMC of SHR and WKY rats were primarily cultured and divided into SHR group, WKY group, Valsartan group, low dose of Chinese medicine group and high dose of Chinese medicine group. The aging of cells were detected by SA?-gal staining, the cell cycle was analyzed by flow cytometry analysis, the levels of MMP-9 and TGF?i were detected by immunocytochemistry and the expression of MLCK was detected by immunofluorescence staining. The mRNA and protein expression of p16, cyclinDl, CDK4, pRb and PPARy were detected by RT-PCR and Western Bloting. Results: Compared with WKY group, VSMC of SHR proliferated significantly, extracts from Panax ginseng, Panax notoginseng (Burk.) and Ligusticum chuanxiong could reduce the SA?-gal expression, arrest the cell cycle in G1 phase, reduce expression of MMP9, TGF?1 and MLCK. The extract could increase the mRNA and protein expression of p16, Rb, PPARy while reducing cyclinDl and CDK4. Conclusion:The extracts from Panax ginseng, Panax notoginseng (Burk.) and Ligusticum chuanxiong can delay vascular aging of SHR rats, which works by cytokines and p16-CyclinD/CDK-RB ways to inhibit VSMC proliferation, migration, reduce the tension of VSMC and the changes of vascular structure caused by high blood pressure.3. Clinical ResearchThe study of relation between the characteristics of TCM syndrome and vascular aging caused by essential hypertensionObjective:To study the relation between the characteristics of TCM syndrome and vascular aging of essential hypertension. Methods:111 patients with essential hypertension were selected as research object and 24 healthy people as control group. The clinical symptoms of hypertension were categorized into different TCM syndrome elements, and the distributions of clinical syndrome of TCM were analyzed. The relation between age, body mass index, pulse pressure, pulse wave velocity, the level of MMP-9, AGEs and TCM syndromes of hypertension were compared. Results: (1) Stagnation of phlegm and yin deficiency is the main syndrome element of hypertension patients, followed by yang hyperactivity, qi deficiency, blood stasis, yang deficiency and blood deficiency; (2) The main clinical syndrome of TCM were syndrome of hyperactivity of yang due to yin deficiency, syndrome of stagnation and obstruction of turbid phlegm, syndrome of intermingled phlegm and blood stasis and syndrome of deficiency of both yin and yang; (3) The patient with syndrome of hyperactivity of yang due to yin deficiency have lower age and BMI than the other groups; (4) The age, PP and PWV of patients with syndrome of intermingled phlegm and blood stasis were significantly increased than the other groups and the levels of MMP-9 and AGEs were also higher than the other groups. Conclusion:Phlegm of TCM plays an important role in the development of essential hypertension, and with the processes of pathogenesis, the relationship of vascular aging and the syndrome of intermingled phlegm and blood stasis become more and more closely.