Study On The Early Treatment Of Reversing Vascular Remodeling And Renin-angiotensin System

PartⅠTheory ResearchEssential Hypertension, or EH, is a multi-cause inherit disease; not onlythere is a high risk in suffering from the disease, but also causes seriouscomplications in heart, brain, and kidney. It causes great threat in personalhealth and huge economical burden in society. It has become a serious publichealth issue between countries.Researches had shown that most victims of EH and SHR already had thickeningin heart blood vessel in early stages; there's also abnormal vascularcontraction activity. This proves the importance and necessity of earlyprevention. "Prevention comes before treatment" is the key point inprevention medicine, and it matches with Chinese traditional medicine saying "To prevent before sickness occur nicely".Hypertension is a noun in modern medicine, as doctors in ancient timedidn't bring up such a name, but mostly use the symptoms to name sickness,such as dizziness, headache, faintness, tinnitus, deafness, ringing in thehead, stroke, amnesia, and insomnia; however faintness and headache were twomain symptoms at that time. It belonged to the area of traditional Chinesemedicine (TCM) type of faintness and liver complaint. Therefore, even ancientdocumentary didn't come up with such names, but the existence of this illnesswas objective. Many TCM documents had mentioned the patho mechanism ofhypertension. The book "Inner Channel" believed, as for internal organs,liver, spleen, and kidney were all responsible for faintness to occur;according to pathology, faintness was caused by deficiency of vital energy.In "Theory of Typhoid", believed one of the cause of dizziness was by damp diet. Four famous doctors in Kim and Yuan Dynasty believed dizziness was causedby the external causes of wind, cold, phlegm, hematoma, and asthenia, but thesymptoms were shown through liver, spleen, and kidney. Modern TCM studyclinical symptoms show that more than 70% victims of hypertension showdizziness and headache. When study on the pulse condition of hypertensionvictims, more than 80% of victims show floating pulse. Multi-level studiesshow liver and fire flaring upward, deficiency in yin and hyperfunction inyang, liver and kidney Yin depletion (vacuity), and disturbance of liver windare the main symptoms of hypertension; damp and blood obstruction are commonlyseem side effects and are closely link to any complications relating tohypertension.Combining with the above findings, we believe that the early symptoms ofhypertension are deficiency of vital energy in liver and kidney, hyperfunctionin liver, and phlegm and blood stasis causing arteries and veins blockage.Therefore we had made Chinese medicine that can regulate the function of liverand kidney and removing sputum and blood stasis, to nourish liver and kidney,treats disorders of the liver into "order", and to get rid of phlegm andhematoma.Renin-angiotensin system, or RAS, is crucial in morbidity of hypertension,holds important effect; it stabilize victim' s blood pressure and inner bodyenvironment stability. It includes Renin, AngiotensionⅡor AngⅡ,angiotensionⅠconverting enzyme or ACE, AT1 and AT2. AngⅡcan cause greatvascular contraction, and it can also increase the number of vascular smoothmuscle cell, or VSMC, making hyperplastic within muscle, to promote expressionof original cancer gene, thickening the wall of blood vessels, narrowing vesselcavity, to play a part in hardening of the arteries. It is well-known thestrongest factors in vessel contractions. In one way, through strong vascularcontraction, raising blood pressure in order to promote the reconstructionof artery, at the same time it can also promote vascular smooth muscle cellto thicken and increasing collagen fibre in order to rebuild blood vessel.ACE is an important constituent in RAS and kinetic released enzyme-kineticsystem, it's an important major enzyme in the formation of AngⅡ; it's mainlycontrolled by genes called ACE. It can catalyze the growth of AngⅡwith strongvascular contraction. It can also extend blood vessel passage and slowlydredges vascular matter kinetic, influencing the metabolism balance of sodium and water within the body system, secretion of aldosterone, vascularcontraction and increasing vascular smooth muscle cell. These are the keyfactors in triggering cardiovascular disease, especially hypertension.Therefore this study uses AngⅡand ACE as in rebuilding vascular mechanismin hypertension.PartⅡExperimental research1. Materials and Methods1.1 HerbsComposition and dosage of RR-formula: raidx rehmanniae preparata 25g,carapax et plastrum testudins 30g (break into pieces,fore-decoct),radixsalviae miltiorrhizae 15g,radix notoginseng 5g,ramulus uncariae cum uncis15g (post-decoct),fructus trichosanthis 15g,radix achyranthis bidentatae15g. The herbs are bought from Guangzhou University of Traditional ChineseMedicine affiliated first hospital TCM pharmacy and appraised as pure crudeherbs by the drug department, The concentration of crude herbs is 1.06g/mlafter being fried and concentrated and preserved in refrigerator at 4centi-degree after being cooled under the common temperature.1.2 The subgroup and administration of ratsSpontaneously hypertensive rat, or SHRwas provided by VictonihwaAnimal Laboratory in Beijing, license number 0048303. The age of these ratswere 12-week-old and they were nicely cleaned. There were 17 male rats weighted250±25g, and 34 female rats weighted 160±25g. First, we gave RR-formula tothese rats before sickness at different time period to observe the overalleffect of reversion of SHR vascular rebuilding. We gave SHR to male and femalerats according to their weight at a 1:2 ratio, randomly divided the rats intofour subgroups: 1. medication was given throughout the whole course (S1-10),2. medications was given only in the first four weeks (S1-4), 3. medicationwas given only on week 5-10 (S5—10), and 4. no medication was given at all(Blank). Pair breeding within the same cage after the tail artery was measured;one male rat and two female rats were locked in the cage. There were 4 cagesin one subgroup. We observed the weight and blood pressures on rats that gavebirth to their offspring; we also compared the blood pressure of theseoffspring, in which medication was given at 6-10 weeks at different periodof time. Weight and the thickness of middle membrane smooth muscle were measured on these offspring, and also the situation on increasing reproduction,to see the changes in artery morphology.At the same time, with the method of blood serum pharmacology to observeChinese medicine to in vitro raise VSMC partial RAS function mechanism,therefore experiment three namely from VSMC AngⅡ, the content and the geneexpression of ACE observes and the blood vessel heavy construction relations.2. FindingsExperimental conclusions shows: there was no significant difference(P＞0.05) in weight and blood pressure when using RR-formula to rats ofdifferent group. If medication was given before hypertension sickness occurred,blood pressure of rats of 6-12 week old was significantly lower than blankgroup (P＞0.01). Among them, blood pressure of S1-10 showed significantdifference every week (P＞0.01). Weight and thickening of middle membranemeasuring in offspring after medication was given showed no significantdifference with blank group. However weight of offspring with medication wasstill heavier than non-medicated group each week; weight of each treatmentgroup showed no significant difference, where S1-10 was better than S1-4 group,showing when giving medication right after birth might be good for the growthof offspring. Although the weight of rats in blank groups was lower than othertreatment groups, but value of middle membrane in main artery was significantlythicker than treatment group. There was statistical meaning when comparingS1-4, S1-10, and blank group; however, although the thickness of middlemembrane in S5-10 was lower than blank group, but showed no statistical meaning.The number of BrdU positive cells in blank group increase noticeably; alsodarker in color, and closer to inner membrane. The number of BrdU positivecells in S1-10 was significantly lower than blank group, and with statisticalmeaning; showing that it was more meaningful if medication was given rightafter the rats were born, in reversing vascular remodeling.Through this research we found: changes of amount in vascular smooth musclein all groups AngⅡ, ACE tended to be time-dependent; AngⅡand ACE takenfromVSMC in week 10 and week 12 was significantly higher than week 1 and week5; when comparing AngⅡ, ACE in week 1/week 5 with S12, showed great significantdifference (P＜0.01). When comparing within groups showed, after using serumwith RR-formula, each group showed lower in AngⅡand ACE than the group usingnormal serum, and it's statistical meaningful (P＜0.01). This means serum with RR-formula can lower amount of AngⅡand the activity of ACE within cells,and this might be the mechanism to lower the fattening of VSMC. At the sametime, serum with RR-formula can control the presentation of ACE gene, showingthe function of lowering hypertension mechanism and to reverse vascularremodeling might be because of Renin-angiotensin system was in action.PartⅢConclusion1. Findings showed that giving RR-formula at different time period before thesickness of hypertension occurred could significantly delayed the rate ofblood pressure to rise; the effect was stable and moderate, and the effectcould be seemed even after the medication had be stopped for two weeks. Thisproved that this method could effectively improved the physique of SHR, andalso explained the rationality of using RR-formula combining with clinicalstudies.2. RR-formula could reduced the thickness of middle membrane of VSM and torestrain the presentation of BrdU, showing this formula had certain degreeof reversing vascular remodeling in hypertension, playing a role in protectingblood vessels. The group with medication given throughout the whole courseshowed the best drug efficacy, explaining that the earlier medication was given,duration of time in drug efficacy tended to last longer. Early intervention(week 1-4) might have an decisive significance in the forming and growth ofvascular smooth muscle middle membrane.3. Serum with RR-formula could lower AngⅡand ACE in VSMC, to restrain thepresentation of ACE gene, showing that formula with the function in loweringblood pressure and reversing vascular remodeling mechanism might be becauseof Renin-angiotensin system was functioning in artery restraining.In combination of above findings, RR-formula was used in this experiment,going through the process of macrocosm and cell culture, we found RR-formulahad stability in lowering hypertension, reversing vascular remodeling, andthrough the experiment of macrocosm and cell cultivation, partly provenRR-formula had the mechanism in lowering hypertension and vascular protection.