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ObjectiveTo observe the effects of modified Chaihu Shugansan（CHSG） and its disassembled formulas on angiotensin-converting enzyme 2 （ACE2）-angiotensin （Ⅰ-Ⅶ） ［Ang （Ⅰ-Ⅶ）］-mitochondrial assembly receptor （MasR） axis in hyperlipidemic rats with myocardial ischemia and depression， and to explore the underlying mechanism of its prevention and treatment of myocardial ischemia and depression. MethodA total of 108 male SD rats were randomly divided into a normal group， a model group， a modified CHSG group （11.7 g·kg-1）， a Quyu Huatan disassembled formula group （4.05 g·kg-1）， a Shugan Xingqi disassembled formula group （3.15 g·kg-1）， a Jianpi Yangxue disassembled formula group （4.5 g·kg-1）， a fluoxetine group （0.001 8 g·kg-1）， a trimetazidine group （0.005 4 g·kg-1）， and a simvastatin group （0.001 8 g·kg-1）， with 12 rats in each group. The hyperlipidemia model with myocardial ischemia and depression was induced with a high-fat diet combined with injection of isoproterenol （ISO） and chronic unpredictable mild stress （CUMS） in rats in the model group and groups with drug intervention for eight weeks. The rats in each group with drug intervention were treated correspondingly by gavage from the first day of modeling， while those in the normal group and the model group received the same amount of normal saline. The behavioral changes of rats in each group were observed by open field test and forced swimming test. Left ventricular fractional shortening （LVFS） and left ventricular ejection fraction （LVEF） were measured by echocardiography. The serum levels of total cholesterol （TC）， triglyceride （TG）， low-density lipoprotein cholesterol （LDL-C）， and high-density lipoprotein cholesterol （HDL-C） were detected by the enzyme-labeled apparatus. Hematoxylin-eosin （HE） staining was used to observe the histomorphological changes of the heart. The serum levels of angiotensin Ⅱ （AngⅡ）， ACE2， and Ang（Ⅰ-Ⅶ） were detected by enzyme-linked immunosorbent assay （ELISA）. The protein and mRNA expression of ACE2 and MasR in the hippocampus and the heart was detected by real-time quantitative polymerase chain reaction （Real-time PCR） and Western blot. ResultCompared with the normal group， the model group showed reduced movement time， distance， and average speed in the central area of the open field （P<0.01）， prolonged immobility time of rats in the forced swimming test （P<0.01）， decreased LVFS and LVEF （P<0.01）， inflammatory exudation and disorderly arranged fiber in heart tissues， elevated serum levels of TC， LDL-C， AngⅡ， ACE2 and Ang（Ⅰ-Ⅶ）， diminished HDL-C （P<0.01）， dwindled mRNA and protein expression of ACE2 in the hippocampus and the heart and MasR in the hippocampus， and up-regulated mRNA and protein expression of MasR in the heart （P<0.01）. Compared with the model group， the modified CHSG group displayed increased movement time， distance， and average speed in the center area of the open field （P<0.01）， shortened immobility time in the forced swimming test （P<0.01）， increased LVFS and LVEF （P<0.01）， relieved heart injury， reduced serum levels of TC， LDL-C， AngⅡ， ACE2， and Ang（Ⅰ-Ⅶ）， elevated level of HDL-C （P<0.01）， up-regulated mRNA and protein expression of ACE2 in the hippocampus and the heart and MasR in the hippocampus， and down-regulated mRNA and protein expression of MasR in the heart （P<0.01）. Each disassembled formula could improve the above indexes to a certain extent （P<0.05， P<0.01）， but the effect of the whole formula was optimal. ConclusionThe modified CHSG and its disassembled formulas have the effects of resisting depression， improving myocardial injury， and reducing blood lipid. Due to the synergistic effects of stasis-resolving/phlegm-eliminating drugs， liver-smoothing/Qi-moving drugs， and spleen-tonifying/blood-nourishing drugs in the formula， the modified CHSG is superior to each disassembled formula in efficacy. Its mechanism may be related to the activation of the ACE2-Ang （Ⅰ-Ⅶ）-MasR axis.
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Objective To systematically assess the relation between angiotensin-I converting enzyme(ACE) gene insertion/deletion (I/D) variation and type 2 diabetic nephropathy (T2DN) onset risk among Chinese population.Methods The related literatures were retrieved from the China National Knowledge Infrastructure (CNKI) and Wanfang Data until June 1st,2016.The RevMan 5.0 was used to conduct the statistical analysis.The merge OR value and corresponding 95% confidence interval(95%CI) were used to assess ACE gene I/D polymorphism and T2DN onset risk.Results Totally 29 papers with 4 357 subjects were included according to the inclusion and exclusion standard,including 2 208 cases of DN and 2 149 cases of T2DM without DN.Meta analysis showed that compared with ACE gene I/D polymorphism I allele,D allele could significantly increase the risk of T2DM patients suffering from DN,the OR value and corresponding 95%CI were 1.44(1.25,1.66);the gene analysis showed that ACE gene I/D polymorphism loci were significantly correlated with DN onset risk in the Asian population.The corresponding relative onset risk OR and 95%CI were 1.42(1.15,1.76) and 1.75(1.46,2.10) in the dominant and recessive genetic model.The Begg′s test showed that the included data had no obvious publication bias existence.Conclusion ACE gene I/D polymorphism is closely correlated with the onset risk of T2DN,and D allele might be a risk genetic factor for DN occurrence in the patients with T2DM.
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Objective:The chemiluminescence immunoassay for AngiotensinⅠ ( AngⅠ ) was developed by the competition method. The renin activity was calculated by the determination of Ang Ⅰ. Methods: AngⅠ antigen was labeled with biotin and fluorescein labeled AngⅠantibody,and anti fluorescein antibody was used to coated with the microporous plate . Results:The standard range of the method was 100-7 800 pg/ml. The assay sensitivity was 24. 3 pg/ml. The intra- and inter-assay coefficients of variance were 5. 6%-8. 7% and 6. 8%-10. 4% respectively. Analytical recovery was 95. 4%-105. 3%. The correlation coefficients between measured and expected values were 0. 999 after serial diluted. Compared with radioimmunoassay kit,the correlative equation was y=0. 95x+235. 70,correlation coefficient was 0. 973. Coated microporous plate and biotin labeled Ang Ⅰ antigen,and fluorescein labeled Ang Ⅰantibody at 37℃ for a week with good stability. Conclusion: The results of the method were in accord with the basic requirements of immunoassay.
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Objective To investigate the effect of Ang-(1-7) on the apoptosis in human umbilical vein endothelial cells (HUVECs) induced by Ang Ⅱ.Methods HUVECs were isolated and cultured.Cultured HUVECs were incubated for 24 h with Ang-(1-7), Ang Ⅱ, Ang-(1-7) A-779, Ang-(1-7) + Ang Ⅱ, A-779 + Ang Ⅱ + Ang-( 1-7), respectively.Cultured HUVECs without incubating stimulator were chosen as controls.The apoptosis of endothelial cells were detected by flow cytometry.Results The apoptosis of endothelial cells in HUVECs were upregulated by AngⅡ ( 10-6 mol/L) (25.60% ±3.17% vs 2.32% ±0.24%, P ＜0.005).Compared with the AngⅡ group, Ang-(1-7) dose-dependently inhibited the apoptosis of endothelial cells in HUVECs ( (20.04% ± 2.21% ,16.04% ± 1.32 %, 10.04% ±2.05,7.79% ±1.50% vs AngⅡ group 25.60% ±3.17%, P ＜0.05 , P ＜0.05).The effects of Ang-(1-7) could be blocked by A-779 (23.37% ±0.75% vs 20.04% ± 2.21%, 16.04% ± 1.32,10.04% ± 2.05% ,7.79%± 1.50%, P ＜ 0.05 ).Conclusion Ang-(1-7) can attenuate the apoptosis of endothelial cells induced by Ang Ⅱ in HUVECs in a dose-dependent manner.The effects of Ang-(1-7) could be blocked by A-779( P＜0.05).
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To explore the relation of angiotensin-converting enzyme (ACE) and angiotensin Ⅱ type 1 receptor (AT1R) gene polymorphism with coronary heart disease (CHD) and the severity of coronary artery stenosis, 130 CHD patients who underwent coronary angiography were examined for the number of affected coronary vessels (≥75% stenosis) and coronary Jeopardy score. The inser- tion/deletion of ACE gone polymorphism and ATIR gene polymorphism (an A→C transversion at nucleotide position 1166) were detected by using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) in CHD patients and 90 healthy serving as controls. The re- sults showed that DD genotype and of ACE were more frequent in CHD patients than that in control group (38.5% vs 14.4%, P<0.001). The frequency of the AT1R A/C genotypes did not differ between the patients and the controls (10% vs 13.1%, P0.05). The relative risk associated with the ACE-DD was increased by ATIR-AC genotype. Neither the number of affected coronary vessels nor the coro-nary score differed among the ACE I/D genotypes (P0.05). But the number of affected coronary vessels and the coronary score were significantly greater in the patients with the AT1R-AC genotype than in those with the AA genotype (P<0.05). In conclusion, DD genotype may he risk factor for CHD and MI in Chinese people, and is not responsible for the development of the coronary artery stenosis. The AT1R-C allele may increase the relative risk associated with the ACE-DD genotype, and may be involved in the development of the stenosis of coronary artery.
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AIM To establish a new bioassay method for estimating the rat angiotensin Ⅰ converting engyme (ACE) activity in vivo by using ramipril (Ram), a ACE inhibitor. METHODS The change in mean arterial pressure induced by angioteuasin Ⅰ (AngⅠ) or bradykinin (BK) was measured before and 1, 24, 48 h after pretreatment with Ram (0 05,0 1 mg?kg -1 ,iv). The ACE activity was expressed by the pressor effect (%)of AngⅠ or the depressor effect (%) of BK. RESULTS The pressor effects of AngⅠ or the supernatant fraction of heart tissue homogenate containing AngⅠ were siginificantly reduced 1 h and 24 h after pretreatment with Ram, but not 48 h after pretreatment with Ram. In comparison of the depressor response of BK vs pressor response of AngⅠ 1 h after pretreatment with Ram,it was shown that the depressor effect of BK is more sensitive than the pressor effect of AngⅠ( P
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Plasma plasminogen activator inhibitor 1 (PAI-1) antigen level in 204 patients with type 2 diabetes mellitus (DM) was higher than that in 60 healthy subjects (P
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The dynamic changes about vasoactive intestinal peptide (VIP), angiotension Ⅰ (AⅠ)and angiotension Ⅱ (A Ⅱ)concentrations in plasma, cerebral spinal fluid (CSF) and brain extracts were studied in the experimental high-velocity missile wound models, which were produced on both hind legs of dogs. The findings were as follows: (1) Plasma VIP, AⅠ and AⅡ concentrations started to increase from 30 rain after the wound and the elevation had been continuing at 4 h after the wound. Their highest points of post-trauma were 2,9, 11.5 and 4.9 times as much as that of pre-trauma. (2) CSF VIP elevated at 4 h after the wound, but AⅠ and AⅡ concentrations reduced tempo tarily at 1 h and 2 h after the wound respectively. (3) The changes of VIP, AⅡ concentrations in the brain extracts tented to be consistent with that of the CSF. The signifcance and machanism of these changes of VIP and AⅡ concentration remain to be clarified.
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The present study was performed to determine whether or not that cardiacrenin-angiotensin system (RAS) plays an important role in volume-overload myocardialhypertrophy. In order to control the increase of circulating RAS, aortacava fistula withnephrectomy (ACF+NT) rats were used in the experiment. The ventricular angiotensin Ⅰ,Ⅱ (Ang Ⅰ,Ⅱ) contents and angiotensin-converting enzyme (ACE) activities were measuredby means of radioimmunoassay and biochemical method. It was shown that in ACF+NTrats, the Ang Ⅱ contents and ACE activities in both left and right ventricles increasedsignificantly despite the plasma Ang Ⅰ, Ⅱ and renin activity remained at a lower level.These results indicate that: (1) Cardiac RAS as a relatively independent system, plays aregulatory role in the development of volume-overload cardiac hypertrophy; (2) The in-crease of Ang Ⅱ content was partly caused by the enhancement of ACE activities; (3) Atleast at the early stage of volume-overload cardiac hypertrophy, the circulating RAS maynot be a necessary promoter.
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We studied the alteration of local and circulat-ing RAS during cardiac hypertrophy caused by ab-dominal aorta constriction (AC, with nephrectomyof left side) and aorta-vena fistula (AVF). Bothventricular and plasma angiotensin Ⅰ and Ⅱ (AngⅠ. Ⅱ), and plasma, renin actvity (RA) weremeasured by means of radioimmunoassay after twoand seven weeks of surgery. Despite the level ofplasma Ang Ⅰ, Ⅱ and RA were not enhanced, theventricular Ang Ⅰ and Ⅱ were increased signifi-cantly in AC rats (v. s. SO group P
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The kinetic changes of the renin-angiotensin-aldosterone system (RAAS) were studied in 54 cases of epidemic hemorrhagic fever (EHF). It was found that plasma angiotensin Ⅱ (A T-Ⅱ) levels began to increase during the febrile period, reached the peak values during the hypotensive or oliguria period, and then dropped gradually during the polyuria period. However, the plasma angiotensin Ⅰ (AT-Ⅰ) levels decreased markedly lower in patients at the hypotensive or oliguria slage than in those at other stages. The plasma aldosterone (ALD) increased, corresponding to AT-Ⅱ during the hypotensive or oliguria period. The above changes were parallel to the severity of the disease and the quantity of BUN. From the results of our studies, it is concluded that the increased plasma AT-Ⅱ concentration and the activation of RAAS are related to hypovolemia and may be the most important factor of acute renal failure. Therefore, it is critical treatment at the early stage of EHF to maintain the equilibrium of body fluids and to resist the effects of activated RAAS.