Prior vasorelaxation enhances diadenosine polyphosphate-induced contractility of rat mesenteric resistance arteries

Low-threshold concentrations of diadenosine polyphosphates (ApnA: Ap3A, Ap4A, Ap5A, Ap6A) or ATP, which at basal vessel tone induce just measurable vasoconstrictions, induce up to ten times enhanced vasoconstrictions of previously relaxed (by acetylcholine or sodium nitroprusside or 8Br₂ cGMP or isoproterenol or levcromakalim) pre-contracted rat mesenteric resistance arteries (MrA) in a microvessel–myograph. These enhanced vasoconstrictions were of similar magnitude for threshold concentrations of all ApnA.Possibly, the low concentrations of ApnA reverse the prior vasorelaxation by inhibiting a common vasorelaxation pathway, but obviously this is not due to inhibition of guanylate cyclase, which has been previously described to be inhibited by ApnA, because the enhanced vasoconstrictions can be observed with guanylate cyclase-independent vasorelaxants (8Br₂ cGMP, isoproterenol or levcromakalim), too. The enhanced vasoconstrictions are endothelium-independent because after mechanical vascular de-endothelialization the results were identical. De-endothelialized vessels, which fail to relax by acetylcholine, showed no enhanced ApnA-induced vasoconstrictions, demonstrating that the mere prior vasorelaxation of the vessel is required to provide the enhanced vasoconstriction by ApnA. Furthermore, the enhanced contractility is not based on a potentiation of the phenylephrine contraction because it equally occurs with other agents used for arterial pre-contraction. Systemically applied ApnA considerably decrease arteriovascular resistance, resulting in hypotension. But here it is demonstrated that a preceding vasorelaxation enables the resistance arteries to generate a strong and persistent ApnA-induced vasoconstriction. Thus, in vivo at very low concentrations ApnA may serve to counteract severe conditions of hypotension (e.g., shock syndrome or anaphylaxis) by the constriction of resistance arteries.     

Arterial relaxation is coupled to inhibition of mitochondrial fission in arterial smooth muscle cells: comparison of vasorelaxant effects of verapamil and phentolamine

Mitochondria are morphologically dynamic organelles which undergo fission and fusion processes. Our previous study found that arterial constriction was always accompanied by increased mitochondrial fission in smooth muscle cells, whereas inhibition of mitochondrial fission in smooth muscle cells was associated with arterial relaxation. Here, we used the typical vasorelaxants, verapamil and phentolamine, to further confirm the coupling between arterial constriction and mitochondrial fission in rat aorta. Results showed that phentolamine but not verapamil induced vasorelaxation in phenylephrine (PE)-induced rat thoracic aorta constriction. Verapamil, but not phentolamine, induced vasorelaxation in high K⁺ (KPSS)-induced rat thoracic aorta constriction. Pre-treatment with phentolamine prevented PE- but not KPSS-induced aorta constriction and pre-treatment with verapamil prevented both PE- and KPSS-induced aorta constriction. Transmission electron microscopy (TEM) results showed that verapamil but not phentolamine inhibited KPSS-induced excessive mitochondrial fission in aortic smooth muscle cells, and verapamil prevented both PE- and KPSS-induced excessive mitochondrial fission in aortic smooth muscle cells. Verapamil inhibited KPSS-induced excessive mitochondrial fission in cultured vascular smooth muscle cells (A10). These results further demonstrate that arterial relaxation is coupled to inhibition of mitochondrial fission in arterial smooth muscle cells.     

Evaluation of triclosan exposures on secretion of pro-inflammatory cytokines from human immune cells

Triclosan (TCS) is widely used in personal hygiene products, such as mouthwash and toothpaste, and is found in human tissues. Interleukin (IL)-1 beta (IL-1β), IL-6, tumor necrosis factor alpha (TNFα), and interferon gamma (IFNγ) are pro-inflammatory cytokines and inappropriately elevated levels of each have been associated with pathologies including rheumatoid arthritis and certain cancers. Here we examine effects of TCS on the secretion of the pro-inflammatory cytokines from human immune cell preparations. TCS at concentrations between 0.05–5 μM consistently increased the secretion of IL-1β, IL-6, and TNFα within 24 h of exposure and the increases often maintained out to 6 days of exposure. TCS also induced increases in IFNγ secretion, however the increases were most consistent after 48 h of exposure rather than within 24 h. Additionally, a role for both p44/42 and p38 MAPK in TCS-stimulated increases in IL-1β was seen in cells from some donors.     

Potential estrogenic activity of triclosan in the uterus of immature rats and rat pituitary GH3 cells

Triclosan (5-chloro-2-(2,4-dichlorophenoxy)phenol; TCS) is used as an antimicrobial agent in personal care, pharmaceutical, industrial, and household products. In this study, we established an in vivo model for screening estrogenic activity of TCS in the uteri of immature rats. In addition, we employed temporarily transfected cells with plasmids containing estrogen response element (ERE) and progesterone (P4) response element (PRE) sequences. We found that uterine weight was significantly increased by 17α-ethinylestradiol (EE) as a positive control and TCS at doses of 7.5, 37.4, and 187.5 mg/kg. In addition, the expressions of calbindin-D_9k (CaBP-9k) and complement C3 (C3) were significantly induced by EE and TCS in the uteri of immature rats, indicating that TCS can induce their expression mediated by estrogenic activity. Co-treatment with steroid antagonists ICI 182,780 (ICI) and RU 486 in conjunction with TCS (37.5 mg/kg) reversed TCS-induced uterine weight and CaBP-9k mRNA and protein expression increases in immature rats. Moreover, ERE and PRE luciferase activity was evaluated in GH3 cells following treatment with TCS. Concentrations of TCS at increasing doses (10⁻⁹, 10⁻⁷, and 10⁻⁵ M) resulted in a significant increase in ERE luciferase activity compared to control; however, no difference was observed in PRE luciferase activity following TCS treatment. To confirm that ER signaling is involved in TCS-induced CaBP-9k expression, we treated GH3 cells with the anti-estrogen ICI, which can block TCS-induced up-regulation of CaBP-9k in these cells. Taken together, these results indicate that TCS has an estrogen-like property, which may be mediated through an ER-involved signaling pathway in both in vivo and in vitro models.     

Urocortin-2 induces vasorelaxation of coronary arteries isolated from patients with heart failure

1. Urocortin-2 (Ucn2) is a vasoactive peptide belonging to the corticotrophin-releasing factor (CRF) family that has potent cardiovascular actions. It has been suggested that Ucn2 participates in the pathophysiology of heart failure. However, little is known about the mechanisms underlying the action of Ucn2 in human coronary arteries. The aim of the present study was to assess the effects of Ucn2 on the vascular tone of human coronary arteries dissected from heart failure patients. 2. Human coronary arteries were dissected from the hearts of patients subjected to orthotopic heart transplantation. Coronary arteries were obtained from 17 patients with heart failure due to dilated cardiomyopathy of ischaemic origin in Stage III-IV of the New York Heart Association classification. Changes in tone were measured in arterial rings using force transducers. 3. Application of increasing concentrations of Ucn2 (5-20 nmol/L) to arterial rings precontracted with agonists induced dose-dependent relaxation of the coronary artery, which was independent of endothelial cell activation. Furthermore, the inhibition of the adenylyl cyclase by MDL-12 (100 nmol/L) and protein kinase A (PKA) by H89 (1 μmol/L) prevented Ucn2-mediated relaxation of coronary artery rings. 4. The results of the present study suggest that, in heart failure patients, Ucn2 could be useful in modulating coronary artery circulation independent of endothelial integrity through mechanisms that involve adenylyl cyclase activation and PKA stimulation. The findings warrant further investigation of the role of Ucn2 in circulatory regulation and its potential therapeutic application in heart disease.     

Echinacoside induces rat pulmonary artery vasorelaxation by opening the NO-cGMP-PKG-BKCa channels and reducing intracellular Ca2+ levels

Aim:

Sustained pulmonary vasoconstriction as experienced at high altitude can lead to pulmonary hypertension (PH). The main purpose of this study is to investigate the vasorelaxant effect of echinacoside (ECH), a phenylethanoid glycoside from the Tibetan herb Lagotis brevituba Maxim and Cistanche tubulosa, on the pulmonary artery and its potential mechanism.

Methods:

Pulmonary arterial rings obtained from male Wistar rats were suspended in organ chambers filled with Krebs-Henseleit solution, and isometric tension was measured using a force transducer. Intracellular Ca²⁺ levels were measured in cultured rat pulmonary arterial smooth muscle cells (PASMCs) using Fluo 4-AM.

Results:

ECH (30–300 μmol/L) relaxed rat pulmonary arteries precontracted by noradrenaline (NE) in a concentration-dependent manner, and this effect could be observed in both intact endothelium and endothelium-denuded rings, but with a significantly lower maximum response and a higher EC₅₀ in endothelium-denuded rings. This effect was significantly blocked by L-NAME, TEA, and BaCl₂. However, IMT, 4-AP, and Gli did not inhibit ECH-induced relaxation. Under extracellular Ca²⁺-free conditions, the maximum contraction was reduced to 24.54%±2.97% and 10.60%±2.07% in rings treated with 100 and 300 μmol/L of ECH, respectively. Under extracellular calcium influx conditions, the maximum contraction was reduced to 112.42%±7.30%, 100.29%±8.66%, and 74.74%±4.95% in rings treated with 30, 100, and 300 μmol/L of ECH, respectively. After cells were loaded with Fluo 4-AM, the mean fluorescence intensity was lower in cells treated with ECH (100 μmol/L) than with NE.

Conclusion:

ECH suppresses NE-induced contraction of rat pulmonary artery via reducing intracellular Ca²⁺ levels, and induces its relaxation through the NO-cGMP pathway and opening of K⁺ channels (BK_Ca and K_IR).     

The 2-nitrate-1,3-dibuthoxypropan, a new nitric oxide donor, induces vasorelaxation in mesenteric arteries of the rat

The reduced availability of nitric oxide (NO) is associated with cardiovascular diseases. Therefore, NO donors such as organic nitrates are useful for the treatment of these disorders. The 2-nitrate-1,3-dibuthoxypropan (NDBP) is an organic nitrate synthesized from glycerin, which the pharmacological effects have not been investigated. In this study we evaluated the vasorelaxant effect induced by NDBP in superior mesenteric artery from rats. In phenylephrine pre-contracted artery rings, NDBP (10(-8)-10(-4)M) elicited concentration-dependent and endothelium-independent relaxation, which were attenuated by hydroxocobalamin-HDX (30μM), a NO extracellular scavenger, and 1-H-[1,2,4] oxadiazolo [4,3-a] quinoxalin-1-one-ODQ (10μM), an inhibitor of soluble guanylyl cyclase (sGC). In addition, the NDBP-induced relaxation was reduced by non-selective K(+) channels blocker KCl (20mM) or selective K(+) channels blockers such as tetraethylammonium-TEA (B(KCa), 1mM), charybdotoxin-ChTX (B(KCa), 100nM), glibenclamide (K(ATP), 1μM) and 4-aminopyridine-4-AP (K(V), 1mM). In preparations with ODQ (10μM) plus TEA (1mM), the response was virtually abolished. In rat smooth muscle cells culture, NDBP (10(-6)-10(-4)M) caused concentration-dependent increases in NO levels. These findings suggest that NDBP causes vasorelaxation through NO generation and activation of the sCG/cGMP/PKG pathway.     

酶消化法培养老龄SD大鼠血管平滑肌细胞

目的：采用酶消化法培养老龄SD大鼠血管平滑肌细胞,为血管疾病,特别是为动脉粥样硬化研究提供大量的原代平滑肌细胞。方法：无菌取老龄SD大鼠主动脉,0.2%Ⅱ型胶原酶消化分离细胞,采用自然纯化、差速贴壁纯化平滑肌细胞,免疫组化鉴定平滑肌细胞α肌动蛋白。结果：免疫组化染色显示细胞纯度在95%以上。结论：酶消化法分离获取SD大鼠平滑肌细胞方法简单,易掌握,采用本方法可稳定获得大量的平滑肌细胞供实验使用。     

Inhibition of alpha-receptor-induced Ca2+ release and Ca2+ influx by Mn2+ and La3+

The influence of Mn2+ and La3+ on alpha-receptor-stimulated Ca2+ movements was examined in arterial smooth muscle of the rabbit aorta. Both cations cause an inhibition of phenylephrine (PE) contractile response which exhibits a different pattern at low and high cation concentrations. At 0.1-1.0 mM inhibition by Mn2+ and La3+ was predominately due to a reduction in Ca2+ influx reflected as inhibition of the slow phase of contraction and reduction in PE-stimulated 45Ca uptake. PE log dose-response curves were shifted to the right in a non-parallel manner by 1 mM Mn2+ such that responses to lower PE concentrations were more inhibited. However, in the presence of 10 mM Mn2+ PE responses are equally inhibited at all PE levels. At 10 mM both Mn2+ and La3+ also inhibited PE-stimulated Ca2+ release resulting in a reduction in both the rapid phase of contraction and in the magnitude of PE stimulation of 45Ca efflux. The effects of Nm2+ (1 or 10 mM) on contraction and 45Ca efflux were rapidly reversible, while the effect of La3+ was not. Inhibition of Ca2+ release by 10 mM Mn2+ and La3+ was not caused by displacement of releasable Ca2+, but appeared to reflect their occupation of a superficially located receptor modulating site. The inhibition of Ca2+ influx by lower concentrations of Mn2+ may illustrate the functional consequence of configurational changes in the alpha 2-form of the receptor which have been recently described at lower concentrations of divalent cations.     

Effects of liposome-entrapped adenosine in the isolated rat aorta

This study examined the effects of adenosine- and adenosine deaminase-loaded liposomes upon the contractile activity of the vascular smooth muscle, using the isolated, de-endothelised rat aorta ring as in vitro model. While control liposomes had no effect, intraliposomal adenosine (5 × 10⁻³ M) induced contraction of the preparation. Intraliposomal adenosine deaminase induced partial relaxation of high K⁺-precontracted rings. The adenosine-induced contraction seems to involve Ca²⁺ influx through L-type channels as an essential component, but protein kinase C may also have a modulatory role.     

鼠结肠平滑肌细胞的分离、培养与鉴定

目的建立体外培养鼠结肠平滑肌细胞的方法。方法应用酶解法分离大鼠的结肠平滑肌细胞,用含10％胎牛血清的DMEM液进行鼠结肠平滑肌细胞的原代培养及传代,并以免疫细胞化学对其进行鉴定。结果新鲜分离的结肠平滑肌细胞呈梭形,核居中。培养24h后细胞开始贴壁,3～5d后开始增殖,14d后细胞密集,呈峰谷样生长。细胞经平滑肌特异性肌动蛋白a-actin鉴定,确定为平滑肌细胞。结论应用酶解法可得到急性分离的鼠结肠平滑肌细胞,用含10％胎牛血清的DMEM液重悬分离得到的结肠平滑肌细胞,经过10d左右的培养可以获得结肠平滑肌细胞。该方法重复性好,效果稳定。     

The effect of forskolin on smooth muscle cells of guinea-pig taenia caeci

Forskolin (0.5-10 microM) caused hyperpolarization and relaxation of the smooth muscle cells of guinea-pig taenia caeci (35 degrees C) as measured with the sucrose-gap method. Membrane conductance, reflected by the amplitude of the electrotonic potential, was not changed during the response. The hyperpolarization could also be evoked in the absence of extracellular calcium, calcium and potassium, or calcium and sodium. Further, the 45Ca2+ efflux from taenia caeci was enhanced by forskolin. The results support the concept that calcium extrusion across the plasma membrane is promoted in the presence of forskolin by stimulation of electrogenic calcium pumping.     

Effects of DDT and triclosan on tumor‐cell binding capacity and cell‐surface protein expression of human natural killer cells

1,1,1‐trichloro‐2,2‐bis(4‐chlorophenyl)ethane (DDT) and triclosan (TCS) are organochlorine (OC) compounds that contaminate the environment, are found in human blood and have been shown to decrease the tumor‐cell killing (lytic) function of human natural killer (NK) cells. NK cells defend against tumor cells and virally infected cells. They bind to these targets, utilizing a variety of cell surface proteins. The present study examined concentrations of DDT and TCS that decrease lytic function for alteration of NK binding to tumor targets. Levels of either compound that caused loss of binding function were then examined for effects on expression of cell‐surface proteins needed for binding. NK cells exposed to 2.5 μM DDT for 24 h (which caused a greater than 55% loss of lytic function) showed a decrease in NK binding function of about 22%, and a decrease in CD16 cell‐surface protein of 20%. NK cells exposed to 5 μM TCS for 24 h showed a decrease in ability to bind tumor cells of 37% and a decrease in expression of CD56 of about 34%. This same treatment caused a decrease in lytic function of greater than 87%. These results indicated that only a portion of the loss of NK lytic function seen with exposures to these compounds could be accounted for by loss of binding function. They also showed that loss of binding function is accompanied by a loss of cell‐surface proteins important in binding function. Copyright © 2012 John Wiley & Sons, Ltd.     

Mechanisms of vasorelaxation induced by N-allylsecoboldine in rat thoracic aorta

N-Allylsecoboldine was shown to be the most effective of several boldine derivatives that were tested for their vasorelaxing effect on the rat aorta. In KCl (60 mmol/l) medium, Ca²⁺ (0.03–3 mmol/l)-induced vasoconstriction was inhibited, concentration-dependently, by N-allylsecoboldine. The IC₅₀ for N-allylsecoboldine was calculated to be about 4 mol/l (for a Ca²⁺ concentration of 1 mmol/1). The vasorelaxant effect on KCl-induced responses was more pronounced at 60 mmol/l KCl than at 15 mmol/1 KCI. Contraction of rat aorta in response to phenylephrine (0.01-100 mol/l) was concentration-dependently inhibited by N-allylsecoboldine and by verapamil (3–30 mol/l), while contraction in response to B-HT 920, serotonin or PGF₂ was not affected. This relaxing effect of N-allylsecoboldine persisted in endothelium-denuded aorta. In cultured A 10 vascular smooth muscle cells, N-allylsecoboldine and verapamil displaced the binding of [³H]-prazosin (K _i values = 0.4±0.2 and 0.6±0.2 mol/l, respectively). The increase of inositol monophosphate caused by phenylephrine in rat aorta was completely suppressed by chloroethylclonidine, but only slightly inhibited by N-allylsecoboldine and by verapamil. Glibenclamide or charybdotoxin did not affect the relaxation induced by N-allylsecoboldine of aortic rings precontracted with phenylephrine. Neither the cGMP nor the cAMP content was changed by N-allylsecoboldine. We conclude that N-allylsecoboldine relaxes the rat aorta by blocking Ca²⁺ channels and that it also has an antagonistic effect at ₁-adrenoceptors. Correspondence to: C.M. Teng at the above address     

Threshold of peroxynitrite cytotoxicity in bovine pulmonary artery endothelial and smooth muscle cells

Peroxynitrite is widely reported as highly cytotoxic; yet recent evidence indicates that at certain concentrations, it can induce pulmonary cell hyper-proliferation and tissue remodelling. This study aimed to establish the threshold concentration of peroxynitrite to induce functional impairment of bovine pulmonary artery endothelial (PAEC) and smooth muscle cells (PASMC). PAEC or PASMC were exposed to solution of peroxynitrite or 3-morpholinosydnonimine (SIN-1). Twenty-four hour cell viability, DNA synthesis, and protein biochemistry were assessed by trypan blue dye exclusion, [³H] thymidine incorporation and western blot analysis, respectively. Threshold concentration of peroxynitrite to significantly impair viability of PAEC and PASMC was 2 μM peroxynitrite. In PASMC and PAEC, low concentrations of peroxynitrite (2 nM–0.2 μM) increased cell proliferation and did not activate p38 MAP kinase. The decrease in DNA synthesis and cell viability caused by 2 μM peroxynitrite was associated with caspase-3 cleavage but not p38 activation. Also, 2–20 μM peroxynitrite significantly activated poly ADP ribose polymerase and stress activated kinase JNK in PAEC. However, the higher concentration of 20 μM peroxynitrite did cause a threefold increase in p38 activation. In conclusion, the threshold for the cytotoxic effects of peroxynitrite was 2 μM; which caused apoptotic cell death independent of p38 MAP kinase activation in pulmonary artery cells.     

The biological activities of phthalate esters on rat gastric muscle

Monoethylhexyl phthalate, at concentrations that can occur in blood stored in plastic bags (0.1–0.5 mg/ml), reduced contractions of rat isolated gastric fundus to PGE₂ and acetylcholine; the diethyl compound was less effective. In contrast, dibutyl phthalate (1 and 10 μg/ml) and, to a lesser extent di-isobutyl phthalate, increased the muscle tone. These results are discussed in relation to blood transfusion, and to structural similarities between phthalates and prostaglandins.     

线粒体动力学蛋白异常表达参与阿德福韦酯致大鼠肾小管上皮细胞的损伤

目的：探讨线粒体动力学相关蛋白drp1和mfn1对阿德福韦酯引起的大鼠肾小管上皮细胞损伤的机制。方法：根据阿德福韦酯给药剂量差异,对大鼠随机分为对照组和低、中、高剂量组;对照组给予生理盐水,其余3组分别给予0.2、0.4、0.8 mg·kg-1·d-1阿德福韦酯灌胃。给药60 d后取出大鼠肾脏,常规石蜡包埋,切片,HE染色观察肾小管上皮的形态学改变;免疫组化检测肾小管上皮中drp1和mfn1蛋白的表达。结果：阿德福韦酯干预的可见嗜酸性凝固坏死物和空泡样变性,近曲小管上皮细胞游离面的刷状缘结构不清,上皮细胞排列较紊乱。在drp1及mfn1的免疫组化中,高、中剂量组与对照组之间蛋白表达的平均光密度值差异有统计学意义（P<0.05）。结论：线粒体动力学蛋白drp1/mfn1可能介导了阿德福韦酯对人肾小管上皮细胞（HK-2）的损伤。     

Antimicrobial agent triclosan is a proton ionophore uncoupler of mitochondria in living rat and human mast cells and in primary human keratinocytes

Triclosan (TCS) is an antimicrobial used widely in hospitals and personal care products, at ~10 mm . Human skin efficiently absorbs TCS. Mast cells are ubiquitous key players both in physiological processes and in disease, including asthma, cancer and autism. We previously showed that non‐cytotoxic levels of TCS inhibit degranulation, the release of histamine and other mediators, from rat basophilic leukemia mast cells (RBL‐2H3), and in this study, we replicate this finding in human mast cells (HMC‐1.2). Our investigation into the molecular mechanisms underlying this effect led to the discovery that TCS disrupts adenosine triphosphate (ATP) production in RBL‐2H3 cells in glucose‐free, galactose‐containing media (95% confidence interval EC₅₀ = 7.5–9.7 µm ), without causing cytotoxicity. Using these same glucose‐free conditions, 15 µm TCS dampens RBL‐2H3 degranulation by 40%. The same ATP disruption was found with human HMC‐1.2 cells (EC₅₀ 4.2–13.7 µm ), NIH‐3 T3 mouse fibroblasts (EC₅₀ 4.8–7.4 µm ) and primary human keratinocytes (EC₅₀ 3.0–4.1 µm ) all with no cytotoxicity. TCS increases oxygen consumption rate in RBL‐2H3 cells. Known mitochondrial uncouplers (e.g., carbonyl cyanide 3‐chlorophenylhydrazone) previously were found to inhibit mast cell function. TCS‐methyl, which has a methyl group in place of the TCS ionizable proton, affects neither degranulation nor ATP production at non‐cytotoxic doses. Thus, the effects of TCS on mast cell function are due to its proton ionophore structure. In addition, 5 µm TCS inhibits thapsigargin‐stimulated degranulation of RBL‐2H3 cells: further evidence that TCS disrupts mast cell signaling. Our data indicate that TCS is a mitochondrial uncoupler, and TCS may affect numerous cell types and functions via this mechanism. Copyright © 2015 John Wiley & Sons, Ltd.     

富含微量元素矿泉水对人血管平滑肌细胞生长增殖的影响

目的研究富含微量元素矿泉水对体外培养血管平滑肌细胞（HVSMCs）生长增殖的影响。方法无菌取12例冠状动脉旁路移植术患者术中废弃的大隐静脉,应用金山矿泉水配制的DMEM培养液（MW—DMEM）和双蒸水配制的DMEM培养液（DDW—DMEM）分别培养平滑肌细胞,分为金川矿泉水组和双蒸水组。观察记录2种培养液培养的HVSMCs游出组织块的时间及细胞呈“峰”-“谷”样时间;建立血管紧张素Ⅱ诱导HVSMCs增殖模型,采用MTT比色法观察2种培养液对AngⅡ诱导HVSMCs增殖抑制作用;应用电镜和免疫组织化学染色法观察鉴定HVSMCs。结果金川矿泉水组培养的HVSMCs贴壁时间为（3．7±0．2）d,双蒸水组为（8．5±0．6）d;金川矿泉水组培养的HVSMCs自贴壁后到长成“峰”-“谷”样形态的时间为（8．3±0．2）d,双蒸水组为（7．1±0．5）d;MTT比色法测定金川矿泉水组培养的HVSMCs的增殖活度明显降低;电镜观察MW—DMEM培养的HVSMCs细胞形态完整,表面有丰富的微绒毛。HVSMCs胞浆内富含肌丝,纵向平行排列,胞内有密体;抗-actin平滑肌肌动蛋白免疫组织化学染色后可见胞浆内染成褐色呈细丝网状。结论富含微量元素和矿物质的金川矿泉水具有修复血管平滑肌细胞损伤和抑制血管紧张素Ⅱ诱导HVSMCs增殖的作用。