Adenovirus-mediated transfer of the atrial natriuretic peptide gene in rat pulmonary vascular smooth muscle cells leads to apoptosis

Atrial natriuretic peptide (ANP) exhibits relaxant and growth-inhibiting effects on vascular smooth muscle cells (VSMCs). To obtain ANP gene expression in VSMCs, we built a recombinant adenovirus containing the ANP cDNA controlled by the adenovirus major late promotor (AdMLP-ANP). After pulmonary VSMC treatment with AdMLP-ANP at a multiplicity of infection ranging from 5 to 100 TCID(50)/cell, immunoreactive ANP was detectable in the cell culture medium at a level that reached 101 +/- 27 pmol/well after 2 days. The newly expressed ANP was biologically active, as evidenced by its ability to induce cyclic guanosine monophosphate accumulation in target cells and to mimic the effect of exogenous ANP (10(-8) to 10(-7) mol/L). Cell growth and survival of AdMLP-ANP-infected cells were decreased and were associated with the promotion of VSMC apoptosis. These effects, which occurred at a multiplicity of infection of 10 to 100 TCID(50)/cell, were observed neither in cells infected with the control adenoviral constructs (AdMLP-betaGAL and AdMLP-gD) nor in cells treated with exogenous ANP (10(-7) to 10(-6) mol/L). These results showing VSMC apoptosis in response to ANP gene expression may have important implications for the prevention of vascular remodeling by gene therapy.     

The effect of atrial natriuretic peptide on cytosolic free calcium in cultured vascular smooth muscle cells

Effect of atrial natriuretic peptide (ANP) on cytosolic free calcium [( Ca2+]i) was studied in monolayers of cultured vascular smooth muscle (VSM) cells loaded with a fluorescent calcium indicator, fura-2. Vasoconstrictive hormones, angiotensin II (AII) and Arg8-vasopressin (AVP) induced initial rapid rises in [Ca2+]i, followed by sustained elevation of [Ca2+]i. ANP (Atriopeptin III 10(-8) M) decreased both the resting level and the sustained elevation of [Ca2+] i induced by AII and AVP. ANP also decreased the rise in [Ca2+]i induced by high potassium (K+) depolarization. AVP-induced initial rapid rise in [Ca2+]i was not inhibited by ANP in the presence or absence of the phosphodiesterase inhibitor, isobutylmethylxanthine 0.1 mM, which has been shown to fully enhance ANP-induced cyclic GMP accumulation. On the other hand, a calcium antagonist, nicardipine, inhibited the high K+-induced rise in [Ca2+]i, whereas it had no effect on not only initial but also sustained rises in [Ca2+]i induced by AVP or AII. These results suggest that ANP has an ability to decrease [Ca2+]i not through inhibition of voltage-sensitive calcium channels, and that neither ANP nor ANP-induced cyclic GMP may affect initial hormone-induced rise in [Ca2+]i. In conclusion, an ability to decrease [Ca2+]i is implicated in ANP-induced relaxation of VSM.     

Renal response to atrial natriuretic peptide in nonedematous sodium-retaining dogs.

ANP administered to cirrhotic dogs or chronic caval dogs with ascites and urinary sodium retention (USR) usually causes heterogeneity of natriuretic response. To assess whether this same phenomenon would occur in the absence of edema, ANP at 100 ng/kg/min was given to dogs before and after the induction of USR by a variety of techniques. Eight dogs were over-diuresed with furosemide over a 2-day period, and 9 dogs were subjected to subacute hemorrhage over a similar period. These dogs were retested with ANP one day later. All 8 dogs given furosemide showed no response to ANP (delta UNa V = 7.4 +/- 4.8 microEq/min), compared to a normal response prior to the diuretic (delta UNa V = 128 +/- 34 microEq/min). The 9 hemorrhaged dogs also responded normally to ANP prior to this manipulation (delta UNa V = 74 +/- 14 microEq/min), but a blunted response post-hemorrhage (delta UNa V = 35 +/- 13 microEq/min). This profile was made up of 5 dogs who responded to ANP (delta UNaV = 62 microEq/min) and 4 who had no response whatsoever (delta UNa V = 3 microEq/min). When 8 dogs were given USR because of continuous mineralocorticoid administration, none responded, but all had a magnified natriuretic response to ANP during the 'escape' phase. Eight dogs were administered minoxidil (10 mg) by mouth daily to induce USR. All 8 dogs responded to ANP (delta UNa V = 93 +/- 6 microEq/min) which was no different from the pretreatment response (delta UNa V = 65 +/- 3 microEq/min).(ABSTRACT TRUNCATED AT 250 WORDS)     

Cyclooxygenase products and atrial natriuretic peptide modulate renal response to endothelin

The effects of porcine endothelin on renal function were investigated in the isolated perfused rat kidney. After control clearance periods, endothelin (80, 120 or 320 pmol) or vehicle was added to the perfusate, and four 10-min experiments followed. Endothelin markedly reduced renal perfusate flow (RPF) as a result of its potent renal vasoconstrictor effect. At the highest but not low doses of endothelin, glomerular filtration rate (GFR) fell disproportionately to the reduction in RPF. Fractional Na excretion was also increased after kidney exposure to endothelin, suggesting an inhibition of tubular Na reabsorption by the peptide. Perfusion with low [Ca] severely blunted the hemodynamic effects of endothelin. Pharmacological blocking of renal cyclooxygenase by indomethacin or ibuprofen caused a further decline in RPF, GFR and absolute but not fractional Na excretion in kidneys challenged with endothelin. Atrial natriuretic peptide increased GFR and filtration fraction (FF), but not RPF, in kidneys previously exposed to 120 pmol endothelin. This was associated with a dramatic diuretic and natriuretic effect. The results demonstrate that 1) endothelin acts directly on the kidney, eliciting hemodynamic, diuretic and natriuretic responses that are dependent on the dose used and partially on the availability of extracellular Ca, 2) the renal effects of endothelin are exacerbated by cyclooxygenase blocking, suggesting that the vasoconstrictor effect of the peptide may be modulated by vasodilatory prostaglandins, and 3) atrial natriuretic peptide counteracts the renal function deterioration induced by endothelin, raising the possibility of an interplay between these vasoactive hormones in the control of renal function.     

Impaired natriuretic response to atrial natriuretic peptide in the isolated kidney of rats with experimental cirrhosis

1. Sodium retention in cirrhosis may be partly attributable to resistance to a putative circulating natriuretic factor. In cirrhosis, plasma concentrations of atrial natriuretic peptide are often increased in the presence of sodium retention. 2. In order to determine whether the kidney of cirrhotic animals may be insensitive to atrial natriuretic peptide, isolated perfused kidneys from six cirrhotic and five control rats were exposed to increasing concentrations of atrial natriuretic peptide. Cirrhosis had been induced by carbon tetrachloride administration. 3. Excretion in vivo of a 2 mmol sodium load, administered by gavage, was impaired in cirrhotic animal for up to 4 h as compared with control animals (4.2 +/- 1.9 vs 34.9 +/- 13.4% P less than 0.05). 4. During perfusion at 110 mmHg in the absence of atrial natriuretic peptide, sodium excretion by isolated kidneys of cirrhotic animals tended to be lower than in control animals, but the difference was not significant (4.93 +/- 1.01 vs 8.41 +/- 1.48 mumol min-1 g-1 kidney weight, P = 0.09). There was a smaller increase in urinary sodium excretion by the kidneys of cirrhotic rats compared with control rats in the presence of atrial natriuretic peptide at 10, 50 and 200 pmol/l (respectively: 0.06 +/- 0.08 vs 1.29 +/- 0.35 mumol/min-1 g-1 kidney weight, P less than 0.02; 0.49 +/- 0.08 vs 1.82 +/- 0.42 mumol min-1 g-1 kidney weight, P less than 0.03; 1.42 +/- 0.16 vs 3.23 +/- 0.73 mumol min-1 g-1 kidney weight, P less than 0.05), but not at 1000 pmol/l.(ABSTRACT TRUNCATED AT 250 WORDS)     

Coronary endothelial dysfunction and impaired microcirculation response to atrial natriuretic peptide in hyperinsulinemia

Endothelial dysfunction occurs in hyperinsulinemia (HI). Coronary microcirculation responses to vasoactive agents are examined in 57 patients with angiographically normal coronary arteries. Patients were divided into 2 groups, 37 with normoinsulinemia (NI) and 20 with HI based on results of a 75-g oral glucose tolerance test. Epicardial artery vasoactivity in response to acetylcholine chloride is measured to assess endothelial function. Coronary microcirculation function is evaluated by intracoronary administration of 50 microg of adenosine triphosphate, 1 mg of isosorbide dinitrate, and 0.05 mg/kg of atrial natriuretic peptide. Epicardial artery vasoconstriction in response to 100 microg of acetylcholine is mildly reduced in HI (P = .04). Coronary flow reserve in response to adenosine triphosphate in NI is similar to that in HI. In NI, the resting mean (SD) peak velocity in response to isosorbide dinitrate (40.7 [10.9] cm/s) vs atrial natriuretic peptide (39.6 [10.9] cm/s) is similar. In contrast, the resting mean (SD) peak velocity in response to atrial natriuretic peptide (31.3 [9.3] cm/s) vs isosorbide dinitrate (43.5 [10.0] cm/s) in HI is statistically significantly blunted (P < .001). Atrial natriuretic peptide may have a pathologic effect on coronary microcirculation even in mild endothelial dysfunction among patients with HI.     

不同运动量游泳运动对大鼠心房肌细胞心钠素颗粒的影响

目的:探讨不同运动量游泳运动对大鼠右心房肌细胞心钠素颗粒的影响,以期为运动对心肌的生理和病理变化研究提供形态学依据。方法:SD大鼠分为安静对照组、不同运动量运动组和力竭运动组。不同运动量运动组进行12周的低、中和高运动量游泳运动,力竭运动组进行1次力竭游泳运动。取右心房肌进行常规透射电镜样品制备,观察心房肌细胞内心钠素颗粒的数量和分布,并统计分析心钠素颗粒大小。结果:安静对照组的右心房肌细胞中心钠素在内质网合成后,经高尔基复合体加工和分泌,形成高尔基分泌小泡,分泌小泡相互融合,形成大的心钠素颗粒。不同运动量运动组的结果表明,随着运动量的增加,心钠素颗粒形成逐渐增加,并迅速分泌到细胞外。力竭运动组的结果表明,力竭运动后即刻,有心钠素颗粒的形成和分泌,而力竭运动后不同时间内,心钠素颗粒被快速分泌到细胞外,而没有增加心钠素颗粒的形成。结论:运动可以诱导心钠素颗粒的形成和分泌,随着运动量的增加,心钠素颗粒的形成和分泌增加。力竭运动后逐渐诱导心钠素颗粒的分泌。     

A renorenal suppression in response to atrial natriuretic peptide in the unilateral ischaemic rat

1. The present study was carried out to investigate the haemodynamic and natriuretic effects of atrial natriuretic peptide (ANP) in normal rats and in rats with unilateral ischaemia. 2. Twenty-four hour unilateral ureter occlusion gave rise to a marked vasoconstriction in the ipsilateral kidney after its release. Intravenous infusion of ANP doubled p-aminohippurate clearance and inulin clearance and elicited massive natriuresis in the hydronephrotic kidney, while in the contralateral kidney these clearance values were decreased and there was a lack of natriuresis. The responses in the latter kidney were also different from those in the normal rat, in which significant natriuresis was elicited by ANP even though a decrease in p-aminohippurate clearance occurred. 3. After the control kidney in the unilateral hydronephrotic rat was denervated either mechanically or by pretreatment with prazosin, ANP induced a natriuresis in both kidneys. Furthermore, the renal denervation prevented the decrease in inulin clearance in the control kidney after ANP administration. 4. These findings suggest that the renal response to ANP may depend on the vascular tone before administration, and that renal nerve activity may modify the effects of ANP on renal haemodynamics and sodium excretion.     

Age-related changes in muscles and joints

Aging muscle and joint changes can have a tremendous impact on the functionality of elderly people with and without disabilities. Studies in animal models have shown some potentially beneficial interventions (eg, gene therapy). Further research is needed to ascertain their benefits in humans. A better understanding of mechanisms by which skeletal muscle and joint changes take place in a geriatric population will be helpful to find reasonable ways to prevent age-related change and improve disability. Although some agents have been reported to have significant positive effects, further studies are needed to determine long-term side effects. More information is needed with respect to the changes in muscles and joints in various disabilities.     

Autocrine and paracrine effects of atrial natriuretic peptide gene transfer on vascular smooth muscle and endothelial cellular growth.

In addition to the atria, recent evidence suggests that atrial natriuretic peptide (ANP) is also synthesized in other tissues. Of particular interest is the location of ANP mRNA in the vessel wall. We and others have shown that exogenously added ANP inhibited the growth of endothelial cells and vascular smooth muscle cells (VSMC) in culture. However, it is not known if the locally synthesized ANP would act similarly. Because cultured endothelial cells and VSMC have lost the ability to express the endogenous ANP gene, we have transfected cells in culture with an expression vector expressing rat ANP and have examined the effects on growth. Cultured endothelial cells transfected with an ANP expression vector synthesized and secreted high levels of ANP. These cells also showed significantly lower rates of DNA synthesis under basal and fibroblast growth factor (FGF)-stimulated conditions. Addition of conditioned medium from endothelial cells transfected with ANP vector to nontransfected endothelial cells resulted in the significant increase in cyclic GMP. Similarly, conditioned media collected from endothelial cells transfected with ANP vector also decreased DNA synthesis in VSMC. Coculture of ANP-transfected endothelial cells with quiescent VSMC showed that released ANP from endothelial cells inhibited DNA synthesis in VSMC. Finally, we examined the autocrine effect of direct transfection of ANP vector into VSMC. Transfection of the ANP vector decreased DNA synthesis in VSMC under basal and angiotensin II-stimulated conditions. Similarly, transfection of the ANP vector resulted in a decrease in the PDGF and serum (5%)-stimulated DNA synthesis of VSMC. These results demonstrate that endogenously produced ANP can exert autocrine and paracrine inhibitory effects on endothelial cell and VSMC growth. In vivo gene transfer of ANP may provide us with the opportunity of gene therapy for vascular diseases in which the abnormalities are vasoconstriction and pathological growth.     

利钾尿肽与心钠素摩尔比值在高血压性脑血管病的临床意义

目的探讨利钾尿肽(KP)与心钠素(ANP)摩尔比值在高血压性脑血管疾病中的意义.方法用放射免疫分析方法测定40例脑血管疾病(CVD)患者血浆KP和ANP含量.结果高血压性CVD患者血浆KP水平(3.72±0.80)μg/L与单纯CVD患者比较显著降低(P<0.01),血浆ANP含量(0.32±0.10)μg/L明显高于单纯CVD组(P<0.01),高血压CVD患者,病程中KP与ANP摩尔比值(4.30±1.13)显著低于对照组(5.32±1.10)及单纯CVD组(5.19±2.30),P均<0.01.结论KP、ANP含量及KP与ANP比值关系的改变在CVD及高血压性CVD病理生理中起着一定的作用.     

Rat vascular tissue contains a neutral endopeptidase capable of degrading atrial natriuretic peptide

Neutral endopeptidase (NEP, EC 3.4.24.11), purified from renal brush border, cleaves the Cys7-Phe8 amide bond of rat atrial natriuretic peptide (ANP), to generate the inactive metabolite (ANP cleaved at the Cys7-Phe8 bond; x-ANP). To determine if NEP contributes to the inactivation of circulating ANP, we investigated the degradation of rat ANP (rANP, 1-28) in the vasculature. Formation of x-ANP from exogenous ANP was studied in a mesenteric arterial preparation by perfusion in a single pass system in the presence and absence of the NEP inhibitors, thiorphan or phosphoramidon. In addition, a purified membrane fraction was prepared from mesenteric arterial homogenate and compared with an equivalent renal membrane fraction. Formation of x-ANP was quantified by a specific immunoassay (ELISA). Renal and vascular membranes shared the same pH optima for x-ANP formation (pH 7.5), although x-ANP generation was considerably greater in renal vs. vascular membranes (31.6 and 0.4 nmol min-1 mg-1 of protein, respectively). Both preparations were inhibited in a similar, dose-dependent manner by phosphoramidon, thiorphan or a polyclonal antibody to NEP. In perfused mesenteric arteries, 1.6 +/- 0.8 pmol of x-ANP were generated from 1 microgram of ANP; this formation was inhibited 51% by 10 microM phosphoramidon or thiorphan. Plasma levels of x-ANP after bolus i.v. administration of rANP in rats, were inhibited (70-80%) by thiorphan at comparable doses to those used in perfused mesenteric arteries. These studies indicate that ANP is degraded in the vasculature by NEP or an "NEP-like" enzyme(s).     

Abnormal rhythmic oscillations of atrial natriuretic peptide and brain natriuretic peptide in heart failure

The purpose of this study was to clarify whether the secretions of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) are pulsatile in patients with chronic heart failure (CHF), and whether the rhythmic oscillations for ANP and BNP are abnormal in patients with CHF. Several reports have shown that ANP and especially BNP are valuable indicators of the prognosis in CHF. Previously, a pulsatile secretion has been described for ANP and BNP in healthy humans and for ANP in CHF patients. More information about the secretion pattern of BNP in heart failure is necessary to increase the clinical usefulness of BNP in patients with CHF. Patients with left ventricular systolic dysfunction and CHF ( n =12) and controls ( n =12) were investigated. Plasma ANP and BNP levels were determined every 2 min during a 2-h period by radioimmunoassay and analysed for pulsatile behaviour by Fourier transformation. All patients and controls had significant rhythmic oscillations in plasma ANP levels, and 11 patients with CHF and 10 controls had significant rhythmic oscillations in plasma BNP levels. The amplitude of the main frequency was considerably higher in patients with CHF than in controls (ANP: CHF, 4.76 pmol/l; controls, 0.75 pmol/l; P <0.01. BNP: CHF, 3.24 pmol/l; controls, 0.23 pmol/l; P <0.001; all values are medians), but the main frequency did not differ significantly between the group with CHF and the control group for either ANP or BNP. Patients with CHF demonstrate pulsatile secretion of ANP and BNP with a much higher absolute amplitude, but with the same main frequency as healthy subjects.     

Pulsatile secretion of atrial natriuretic peptide and brain natriuretic peptide in healthy humans.

Both atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) are involved in sodium and water homoeostasis in healthy humans. The plasma concentrations of the natriuretic peptides can be used to differentiate between dyspnoea of cardiac and pulmonary origin, and the degree of elevation of the peptide levels in the plasma in heart failure is a measure of the severity of the disease. However, the patterns of secretion of ANP and BNP are not clear either in healthy humans or in patients. The purpose of the present study was to test the hypotheses that both ANP and BNP are secreted in pulses in healthy humans, and that this phenomenon can be revealed by determination of ANP and BNP in peripheral venous blood samples. In 12 healthy subjects, blood samples were drawn every 2 min through an intravenously inserted plastic needle over a period of 2 h. Plasma concentrations of ANP and BNP were determined by RIAs, and the results were analysed for pulsatile behaviour by Fourier transformation. Pulsatile secretion of ANP was seen in 10 out of 12 subjects [nu=0.028 min(-1) (median; range 0.013-0.047 min(-1)), i.e. a pulse of ANP with an interval of 36 min (range 21-77 min)]. Pulsatile secretion of BNP was seen in nine out of 12 patients [nu=0. 021 min(-1) (range 0.013-0.042 min(-1)), i.e. a pulse of BNP with an interval of 48 min (range 24-77 min)]. The main conclusion is that the secretion patterns of both ANP and BNP are pulsatile in most healthy humans. Consequently, it is important to study whether pulsatile secretion also occurs in heart failure in order to obtain the most informative predictive values both in the differential diagnosis of dyspnoea and in the evaluation of the severity of the disease.     

Downregulation of natriuretic peptide clearance receptor mRNA in vascular smooth muscle cells by angiotensin II

Angiotensin II can downregulate atrial natriuretic peptide binding to rat vascular smooth muscle cells (VSMCs), but the mechanism is not known. Because protein kinase C (PKC) mimetic phorbol myristate acetate (PMA) can destabilize natriuretic peptide clearance receptor (NPR‐C) mRNA and angiotensin II activates several PKC isoforms in VSMCs, we hypothesized that angiotensin II treatment decreases NPR‐C mRNA stability and exerts this effect through PKC. This study demonstrated that angiotensin II induced time‐ and concentration‐dependent downregulation of NPR‐C, which was completely inhibited by an angiotensin II type I receptor blocker losartan. NPR‐C mRNA disappearance rate over 6 h was nearly doubled by exposure of VSMCs to 100 nm angiotensin II, compared with that observed after inhibition of RNA synthesis alone. However, this response to angiotensin II was undiminished by the PKC inhibitor chelerythrine, or by depletion of PKC by prior exposure of cells to PMA for 48 h. Inhibitors of tyrosine kinases, phospholipase C, or mitogen‐activated protein kinase kinase also failed to reverse the angiotensin II effect. We conclude that at least two distinct proximal signaling pathways, one involved and one independent of phorbol ester‐sensitive protein kinase C, lead to downregulation of NPR‐C gene expression by destabilizing its mRNA.     

Age-related changes in fatty acid composition in muscles

Changes in lipid metabolism in muscles are closely related to insulin resistance. Insulin resistance is impaired during aging in hereditary hypertriglyceridemic rats. However, changes in the membrane fatty acid composition during aging are poorly understood. The aim of the study was to describe the fatty acid (FA) composition of muscle phospholipids in the diaphragm and in the m. soleus in relation to aging and insulin resistance. Female hereditary hypertriglyceridemic rats (HHTg) have been shown to have elevated levels of serum triglycerides. Fasting and post-load blood glucose and insulin concentrations were higher in HHTg lines as compared with controls of the same age. The most important changes observed in the m. soleus in the HHTg line included decreased saturated FA between 3 and 14 months of age (42.79 +/- 2.30 vs. 28.85 +/- 2.57mol.%, p<0.01) and increased polyunsaturated FA n-6 (27.99 +/- 1.66 vs. 39.37+2.29 mol.%, p<0.01). In the diaphragm we noted, in normotriglyceridemic controls, increased proportions of monounsaturated FA (5.73 +/- 0.80 vs. 11.02 +/- 0.75mol.%, p<0.001), while these proportions were decreased in HHTg rats (13.50 +/- 0.83 vs. 10.46 + 0.61 mol.%, p < 0.05). Age-related changes in muscle phospholipids seem not to be the key to explaining insulin resistance.     

Impaired vasodilator responses to atrial natriuretic peptide in essential hypertension

BACKGROUND: Atrial natriuretic peptide (ANP) has vasodilating and diuretic/natriuretic properties, both of which contribute to lower blood pressure. These effects are mediated by binding of ANP to a cell-surface receptor [type A guanylyl cyclase (GC-A)]. It has been demonstrated by studies in monogenetic mouse models that the ANP/GC-A system participates in the maintenance of blood pressure homeostasis. METHODS: In male patients with essential hypertension (EH; n = 36) as the only cardiovascular risk factor and normotensive controls (n = 12), blood flow was measured in the forearm circulation in response to i.a. infusion of synthetic human ANP, acetylcholine, orciprenaline, and sodium nitroprusside by strain-gauge venous plethysmography. In blood samples, cyclic guanosine'5-monophosphate (cGMP) and ANP concentrations were measured at resting conditions and during exogenous ANP infusion. In 200 patients with EH, genomic DNA was screened for an inhibitory deletion mutation of the GC-A gene, which has been recently linked to EH in a Japanese cohort. RESULTS: The vasodilatations in response to ANP and acetylcholine were impaired in the forearm circulation of patients with EH, whereas the responses to orciprenaline and nitroprusside were preserved. Plasma ANP and cGMP concentrations were increased in the patients with EH both at resting conditions and during ANP infusion; the resting plasma cGMP levels correlated significantly with the plasma ANP levels (r = 0.68). A specific deletion mutation of the GC-A gene did not account for the diminished relaxant effects of ANP in our study population. CONCLUSIONS: The vascular ANP/GC-A pathway is altered in patients with EH, in addition to the known defects on the nitric oxide/cGMP pathway. Attenuation of the vasodilative responses to ANP suggests impaired receptor or postreceptor responsiveness of GC-A. It is possible that this dysfunction participates in the pathomechanism of EH.     

微囊化转心房肽基因细胞分泌心房肽近日节律的体外研究

目的 研究人心房肽 (hANP) cDNA转染细胞的微囊化技术中心房肽分泌的近日节律;通过人工调控,改变心房肽分泌的近日节律 ,达到有效治疗高血压或心力衰竭的目的. 方法 将人心房肽基因( cDNA)转染入中国仓鼠卵巢细胞( Chinese hamster ovary,CHO),然后将细胞包被在聚己内酯管内形成微囊,并检测转基因 CHO细胞长时间存活情况和分泌的心房肽.检测微囊化转基因细胞分泌心房肽的生物节律,并用褪黑素进行调整. 结果 在培养期间,长度 20 mm,直径 2 mm的聚己内酯管内转染 CHO细胞在 2 mL培养基中 24 h分泌的心房肽水平可达 210.3 ng/L;心房肽的分泌呈近日节律性变化,日间高,而夜间低;近日节律变化的时相是 415,用褪黑素处理,近日节律的时相移至 755. 结论 心房肽 cDNA转染的 CHO细胞包被在聚己内酯管内并能向管外分泌心房肽,将来可能适于植入人体.此项研究为心房肽治疗高血压或心力衰竭提供了一条新途径.