钙稳态失衡在氯化镧诱导MDCC-MSB1细胞凋亡中的作用

为研究钙稳态失衡在LaCl3诱导MDCC-MSB1细胞凋亡中的作用,将MDCC-MSB1细胞常规培养于RPMI-1640培养液中,加入终浓度为0.5、1.0、1.5、2.0、2.5、3.0、3.5和4.0mmol/L的LaCl3,继续培养24h后,应用MTT法检测细胞增殖抑制率,DNA Ladder法和TUNEL法检测细胞凋亡,以Fura-2为荧光探针检测细胞内[Ca2+]i的变化.在LaCl3浓度为0.5～4.0 mmol/L范围内,细胞的增殖抑制率增加,细胞凋亡数量和细胞内[Ca2+]i呈升高趋势,并呈剂量-效应关系.结果表明,LaCl3能抑制MDCC-MSB1细胞的增殖,并可能通过改变[Ca2+]i而诱导其发生凋亡.     

钙稳态失衡在氯化镧诱粕CC-MSB1细胞凋亡中的作用

为研究钙稳态失衡在LaCl3诱导MDCC-MSB1细胞凋亡中的作用,将MDCC-MSB1细胞常规培养于RPMI-1640培养液中,加入终浓度为0．5、1．0、1．5、2．0、2．5、3．0、3．5和4．0mmol／L的LaCl3,继续培养24h后,应用MTT法检测细胞增殖抑制率,DNA Ladder法和TUNEL法检测细胞凋亡,以Fura-2为荧光探针检测细胞内[Ca^2＋]i的变化。在LaCl3浓度为0．5-4．0mmol/L范围内,细胞的增殖抑制率增加,细胞凋亡数量和细胞内[Ca^2＋]i呈升高趋势,并呈剂量一效应关系。结l果表明,LaCl3能抑制MDCC—MSB1细胞的增殖,并可能通过改变[Ca^2＋]i而诱导其发生凋亡。     

三氧化二砷诱导鸡MD肿瘤细胞凋亡及对细胞内钙离子浓度的影响

为探讨As2O3诱导MD肿瘤细胞株凋亡的机制,采用MTT法检测As2O3对MDCC-MSB1的抑制作用.采用荧光显微镜观察细胞形态学变化,DNA Ladder法检测细胞凋亡,Fura-2/AM负载双波长荧光分光光度计检测细胞内游离钙离子浓度([Ca2 ]i)变化,观察A2O3对鸡MD肿瘤细胞株MDCC-MSB1的影响.结果显示:As2O3能抑制鸡MD肿瘤细胞株MDCC-MSB1的生长,呈剂量依赖关系(P<0.05或P<0.01);在荧光显微镜下可见As2O3作用后MDCC-MSB1细胞呈现典型的凋亡特征,并出现典型的DNA Ladder,双波长荧光分光光度计检测结果显示:As2O3作用后的MDCC-MSB1细胞内[Ca2 ]i升高(P<0.05或P<0.01),呈剂量依赖关系.As2O3能明显抑制鸡MD肿瘤细胞株MDCC-MSB1细胞的生长,并诱导其发生凋亡,细胞内[Ca2 ]i高可能是其诱导细胞凋亡的机制之一.     

钙稳态失衡在锰致体外培养鸡胚脑神经细胞凋亡中的作用

旨在探讨钙稳态失衡在锰致体外培养鸡胚脑神经细胞凋亡中的作用。以体外培养鸡胚脑神经元为研究对象,在含终浓度为0、1.5、22、.5 mmol.L-1MnCl2的DMEM培养液中培养24 h,应用流式细胞仪检测磷脂酰丝氨酸(PS)和线粒体膜电位(ΔΨm),琼脂糖凝胶电泳法(DNA Ladder)检测细胞染色质的断裂,以Fura-3/AM为探针检测细胞内游离钙离子浓度([Ca2+]i),用实时定量PCR法检测细胞内CaMmRNA的表达水平。结果表明,随着MnCl2浓度的升高,ΔΨm呈降低趋势,PS膜外翻增加,细胞染色质DNA发生损伤,产生明显的梯形条带,细胞内[Ca2+]i呈升高趋势,CaMmRNA的表达水平下降。结果提示,过量锰通过抑制CaM活性,引起神经元内[Ca2+]i升高,膜通透性发生改变,ΔΨm降低,从而导致鸡胚脑神经元发生凋亡。     

镉致大鼠大脑皮质神经细胞凋亡的机理及N-乙酰半胱氨酸的保护作用

对神经细胞用不同浓度(0、5、10、20 μmol/L)醋酸镉染毒和N-乙酰半胱氨酸(NAC)(100μmol/L)进行保护,通过流式细胞仪检测了细胞凋亡率、[Ca2+]i和活性氧(ROS)水平.结果表明,与对照组相比,各染毒组细胞凋亡率、细胞内[Ca2+]i和ROS水平明显升高,呈剂量-效应关系,除5 μmol/L组ROS水平外,均差异极显著(P<0.01).添加NAC组与相应染毒组比较,神经细胞凋亡率有降低趋势,但差异不显著(P>0.05);10、20μmol/L组细胞内[Ca2+]i和ROS水平显著降低(P<0.05或P<0.01).表明镉可诱导神经细胞凋亡,其机理可能与细胞内Ca2+和ROS水平升高有关,NAC对其有一定的保护作用.     

乐果诱导大鼠肝胞凋亡的分析

为研究乐果对大鼠肝细胞凋亡的影响,将24只SD大鼠分成对照组和3个染毒组,分别按体质量以0、1、6和30 mg·kg-1的剂量灌服乐果,连续灌服30 d后,观察肝脏组织学和超微结构变化.同时,通过给大鼠肝细胞培养液中加入乐果(染毒终浓度分别为0、3、10、30、100和300 μmol·L-1),染毒12、24 h后,Annexin V/PI双染法检测肝细胞凋亡率;分别用Fluo-2/AM、双氢-乙酰乙酸二氯荧光黄(DCFH-DA)和罗丹明123检测细胞内Ca2+浓度、活性氧(ROS)和线粒体膜电位(△ψm)变化.结果表明,组织学和超微结构检查显示肝细胞脂肪变性、凋亡等.肝细胞染毒12和24 h后,细胞凋亡率明显升高,除3 μmol·L-1组外,与对照组相比差异显著(P<0.05或P<0.01),且呈时间-剂量效应;3μmol·L-1组细胞内Ca2+浓度极显著高于对照组(P<0.01),之后随染毒剂量的增加,细胞内Ca2+浓度逐渐下降;细胞内ROS水平在3～100μmol·L-1范围内随染毒剂量的增大和染毒时间的延长而升高,而在300 μmol·L-1组略有下降,除3μmol·L-1组外,与对照组相比均差异极显著(P<0.01);△ψm除24 h高剂量染毒组(300 μmol·L-1)外均出现持续下降,30～300 μmol·L-1组均小于对照组(P<0.01).本研究表明低剂量乐果染毒可诱导肝细胞发生凋亡,细胞内Ca2+、ROS和△ψm可能参与了这一过程.     

莲草多糖和黄酮对MDCC-MSB1细胞凋亡及其Bcl-2mRNA表达的影响

为了探讨莲草多糖和黄酮对诱导鸡MD肿瘤细胞凋亡的作用效果及其可能的作用机制,试验以鸡MDCC-MSB1细胞系为研究对象,通过观察细胞形态变化,分析细胞DNA片段化及半定量RT-PCR法检测莲草多糖和黄酮在安全浓度范围内对鸡MD肿瘤细胞凋亡及Bcl-2 mRNA表达的影响。结果表明,莲草多糖和黄酮能诱导MDCC-MSB1细胞凋亡,并降低MDCC-MSB1细胞内Bcl-2 mRNA的表达水平,致使细胞抗凋亡能力下降,进而发挥其药物作用。     

DSS诱导血管内皮细胞ECV304凋亡及机制研究

为了研究葡聚糖硫酸钠(DSS)诱导血管内皮细胞:ECV304增殖和凋亡的影响。体外培养血管内皮细胞ECV304,采用不同浓度DSS诱导细胞,流式细胞术分析细胞凋亡、活性氧(ROS)、线粒体膜电位(MMP)、细胞周期;WST-1法检测细胞增殖抑制率。结果显示,随着DSS浓度的增加细胞凋亡率增加,并导致细胞GOG1期阻滞,细胞内活性氧含量减少,同时线粒体膜电位下降,细胞增殖抑制率明显升高。结果表明,DSS可以抑制血管内皮细胞ECV304增殖并诱导细胞凋亡。     

磷对成骨细胞活性[ Ca2+]i及细胞周期的影响

在体外培养4 dSD大鼠成骨细胞(OB)的基础上,添加不同浓度磷(0、1、2、4 mmol/L)作用48 h后,观察细胞超微结构,测定钙离子浓度([Ca2+]i);作用36 h后,检测OB细胞周期.磷作用48h后,4 mmol/L磷组OB内线粒体减少,总蛋白含量随磷浓度的增加而减少,4 mmol/L磷组显著低于对照组、1 mmol/L磷组和2 mmol/L磷组(P＜0.05);OB内[Ca2+];浓度在磷各组均增加(P＜0.01),2 mmol/L磷高于1 mmol/L磷(P＜0.05),4 mmol/L磷高于1 mmol/L磷(P＜0.01);磷对细胞周期变化不明显,仅1 mmol/L磷抑制细胞滞留在S期(P＜0.05).表明,添加磷作用48 h后,均促进OB内[Ca2+]i沉积,促进OB的体外矿化,4 mmol/L磷抑制OB的代谢活性,促进OB提前成熟,利于新骨的形成.     

内质网途径在硫酸黏菌素致PC12细胞凋亡中的作用

为了探究内质网途径在硫酸黏菌素诱导PC12细胞凋亡中的作用。取对数生长期PC12细胞,用含0、62.5、125、250μg/m L硫酸黏菌素的DMEM培养液作用24 h,透射电镜观察PC12细胞凋亡,Western blot法检测Gn RP78、caspase-12、Caspase-3蛋白表达量,Fluo-3/AM检测细胞内[Ca~(2+)]i浓度变化,钙测定试剂盒检测细胞外[Ca~(2+)]i浓度变化。与对照组相比,125、250μg/m L硫酸黏菌素剂量组PC12细胞出现典型凋亡现象,使PC12细胞内[Ca~(2+)]i浓度,Gn RP78、caspase-12、Caspase-3蛋白表达量显著升高(P0.01);细胞外[Ca~(2+)]i浓度显著降低(P0.01),具有剂量-效应关系。说明通过内质网途径介导的PC12细胞凋亡可引发硫酸黏菌素的神经毒性。     

Mechanisms of acetylcholine-induced vasorelaxation in high K+-stimulated rabbit renal arteries

To characterize the mechanisms of acetylcholine (ACh)-induced vasorelaxation in rabbit renal arteries precontracted with high K+ (100 mM), muscle tension and cytosolic free Ca2+ concentration ([Ca2+]i) were measured simultaneously in the fura-2-loaded arterial strips. In the artery with endothelium, high K+ increased both [Ca2+]i and muscle tension. Addition of ACh (10 microM) during high-K+ induced contraction significantly relaxed the muscle and induced additional increase in [Ca2+]i. In the presence of NG-nitro-L-arginine (L-NAME, 0.1 mM). ACh increased [Ca2+]i without relaxing the muscle. In the artery without endothelium, high K+ increased both [Ca2+]i and muscle tension although ACh was ineffective, suggesting that ACh acts selectively on endothelium to increase [Ca2+]i. 4-DAMP (10 nM) or atropine (0.1 microM) abolished the ACh-induced increase in [Ca2+]i and relaxation. However, pirenzepine (0.1 microM), AF-DX 116 (1 microM) and tropicamide (1 microM) were ineffective. The ACh-induced increase of [Ca2+li and vasorelaxation was significantly reduced by 3 microM gadolinium, 10 microM lanthanum or 10 microM SKF 96365. These results suggest that, in rabbit renal artery, ACh-evoked relaxation of 100 mM K+-induced contractions is mediated by the release of endothelial NO. ACh may stimulates the M3 subtype of muscarinic receptor in the endothelial cells, resulting in the opening of the nonselective cation channels followed by an increase of [Ca2+]i and stimulation of NO synthase.     

鸡TGFβ1对MDCC-MSB1细胞增殖、凋亡、迁移与侵袭的影响

旨在探讨鸡TGFβ1对MDCC-MSB1细胞增殖、凋亡、迁移与侵袭的影响。作者将构建的鸡TGFβ1过表达载体、干扰表达载体以及相应阴性对照转染MDCC-MSB1细胞,然后检测转染后各组细胞鸡TGFβ1的表达水平、细胞增殖能力、细胞周期与凋亡,细胞的迁移与侵袭能力。结果显示,与相应阴性对照相比,转染TGFβ1过表达质粒可显著上调MDCC-MSB1细胞的TGFβ1表达水平,显著抑制MDCC-MSB1细胞的增殖,且使G1期细胞增加、S和G2期细胞减少,同时增加细胞凋亡率,降低细胞的迁移与侵袭能力;转染TGFβ1干扰表达质粒可显著下调MDCC-MSB1细胞的TGFβ1表达水平,显著促进MDCC-MSB1细胞的增殖,G1期细胞减少、S和G2期细胞增加,同时降低细胞凋亡率,增加细胞的迁移与侵袭能力。结果表明,鸡TGFβ1可抑制MDCC-MSB1细胞增殖、迁移与侵袭,促进其凋亡。     

Effect of Gallus TGFβ1 on the Proliferation,Apoptosis, Migration and Invasion of MDCC-MSB1 Cells

This study aimed to detect the effect of Gallus TGFβ1 on the biological behavior of MDCC-MSB1 cells. MDCC-MSB1 cells were transiently transfected with Gallus TGFβ1 overexpression vector, interference expression vector, and the corresponding negative control. Then, the expression of Gallus TGFβ1, the cell proliferation, the cell cycle and apoptosis, the migration and invasion of each transfection groups were examined. Results showed that compared with the corresponding control, the MDCC-MSB1 cells transfected with overexpression vector of Gallus TGFβ1 could up-regulate the expression level of TGFβ1, the proliferation of MDCC-MSB1 cells was significantly inhibited, G1 phase cells were increased, S and G2 cells were decreased, the apoptosis rate of the cells was increased, the migration and invasion ability were decreased.However,the MDCC-MSB1 cells transfected with the interference expression vector of TGFβ1 significantly down-regulated the expression level of TGFβ1, cell proliferation was improved,G1 phase cells were decreased, S and G2 cells were increased, the cell apoptosis was decreased, the migration and invasion ability was increased. The results showed that Gallus TGFβ1 could inhibit the proliferation, migration and invasion of MDCC-MSB1 cells, and promote their apoptosis.     

Photodynamic stimulation causes sustained increase in intracellular calcium concentration in cells of small cell lung carcinoma

Photodynamic agents, due to their selective uptake by tumor cells and photon-dependent selective activation, have immense implications for cancer treatment. The present study provided direct evidence that the photon activation of chloro-aluminum phthalocyanine sulphonate (A1PcS4) in the presence of extracellular Ca2+ caused a rapid increase followed by a sustained increase in intracellular concentration of calcium ion ([Ca2+]i) in a small cell lung carcinoma (SCLC) cell line, SBC-3. The [Ca2+]i increase by photodynamic stimulation was completely inhibited by the removal of extracellular Ca2+ and reintroduction of extracellular Ca2+ immediately led to a rapid elevation of [Ca2+]i. However, the increase was not inhibited by application of Ni2+, nifedipine, or SK&F 96365, a receptor-mediated and voltage-dependent Ca2+ entry blocker. The photosensitizer A1PcS4 alone or light alone (4 min) had no effect on [Ca2+]i. Cytotoxicity examination by trypan blue exclusion test, however, suggested photodynamic stimulation-induced cell injury which was observed in both the presence and the absence of extracellular Ca2+. These results indicate that [Ca2+]i increase may not be mandatory for photodynamic stimulation-induced cell injury. Whether [Ca2+]i increase can accelerate, at least in part, cell death under the physiological condition, whether the mechanism(s) of cell death can be different in the presence and the absence of extracellular Ca2+, and whether [Ca2+]i increase can be totally unrelated to cell death await further work.     

Digoxin- and monensin-induced changes of intracellular Ca2+ concentration in isolated guinea-pig ventricular myocyte

This study was undertaken to determine the possible mechanisms of actions of monensin and digoxin by using isolated guinea-pig ventricular myocytes. Since Ca2+ is the major signal for triggering contraction of cardiac muscle, the objective of this study was to determine whether monensin and digoxin affect the [Ca2+]i of cardiac myocytes and if so is this effect due to an increase in [Na+]i. Three different concentrations of digoxin (0.3, 1 and 3 micromol/l) and three different concentrations of monensin (0.3, 1 and 3 micromol/l) were used. Each treatment was monitored for two hours by using computerized fluoroscopy. Both digoxin and monensin increased the [Ca2+]i and accelerated the onset time of [Ca2+]i increase in a dose-dependent manner. Normal myocytes (loaded with fura-2 for 30 min before the treatment) were also compared with 'weakened' myocytes (loaded with fura-2 for 3 h before the treatment to create a 'weakened' condition). It was found that although 0.3 micromol/l monensin and digoxin did not change the [Ca2+]i in normal myocytes, they increased the [Ca2 +]i in 'weakened' myocytes. Finally, a Na+-free medium was used to demonstrate the effect of [Na+]o on both monensin- and digoxin-induced increases in [Ca2+]i. It was found that digoxin did not increase the [Ca2+]i in the Na+-free medium. Although monensin increased the [Ca2+]i in the Na+-free solution, this increase was not as large as in the Na+-containing medium. The results of the study led to the conclusion that the positive inotropic effect of digoxin depends on [Na+]o. However, monensin increases [Ca2+]i in Na+-dependent and -independent ways. An addition conclusion was that 'weakened' myocytes are more sensitive to the monensin and digoxin treatment than normal myocytes.     

Nitric oxide stimulates IP3 production via a cGMP/ PKG-dependent pathway in rat pancreatic acinar cells

In an attempt to explore the functioning of nitric oxide (NO) in pancreatic exocrine cells, we have recently obtained several lines of circumstantial evidence indicating that one of molecular targets of NO is phospholipase C (PLC), the activation of which leads to an increase in the cytosolic Ca2+ concentration ([Ca2+]i) via inositol 1, 4, 5-trisphosphate, IP3. However, whether IP3 is actually produced by NO has not yet been substantiated. The present study was therefore designed to directly measure the intracellular IP3, concentration ([IP3]i) for better understanding of the underlying mechanisms with the help of pharmacological tools. [IP3]i was measured using a fluorescence polarization technique (HitHunter). We obtained the following results: 1) varying concentrations of an NO donor, sodium nitroprusside (SNP), elevated [IP3]i, 2) this elevation was completely inhibited in the presence of the soluble guanylyl cyclase (sGC) inhibitor, 1H-[1, 2, 4] oxadiazolo [4, 3-a] quinoxalin-1-one (ODQ), 3) varying concentrations of the cGMP analogue, 8-Br-cGMP, also increased [IP3]i, 4) the cGMP analogue-induced IP3 production was abolished by pretreatment with either a PLC inhibitor, U73122, or a G-protein inhibitor, GP2A, and 5) KT5823, a potent and highly selective inhibitor of cGMP-dependent protein kinase G (PKG), also abolished the IP3 production induced by 8-Br-cGMP. These results suggest that the NO-induced [Ca2+]i increase is triggered by an increase in [IP3]i located downstream from intracellular cGMP elevation. In this intracellular pathway, each sGC, cGMP-dependent PKG, G-protein and PLC were suggested to be involved. The present work provides new insights into the intracellular signaling accelerated by NO. NO triggers a [Ca2+]I increase via cGMP and IP3 in pancreatic acinar cells.     

Homeostasis of intracellular Ca2+ in equine chondrocytes: response to hypotonic shock

REASONS FOR PERFORMING STUDY: Ca2+ homeostasis in articular chondrocytes affects synthesis and degradation of the cartilage matrix, as well as other cellular functions, thereby contributing to joint integrity. Although it will be affected by mechanical loading, the sensitivity of intracellular Ca2+ concentration ([Ca2+]i) in equine articular chondrocytes to many stimuli remains unknown. HYPOTHESIS: An improved understanding of Ca2+ homeostasis in equine articular chondrocytes, and how it is altered during joint loading and pathology, will be important in understanding how joints respond to mechanical loads. METHODS: [Ca2+]i was determined using the fluorophore fura-2. We examined the effects of hypotonic shock, a perturbation experienced in vivo during mechanical loading cycles. We used inhibitors of Ca2+ transporters to ascertain the important factors in Ca2+ homeostasis. RESULTS: Under isotonic conditions, [Ca2+]i was 148 +/- 23 nmol/l, increasing by 216 +/- 66 nmol/l in response to reduction in extracellular osmolality of 50%. Resting [Ca2+]i, and the increase following hypotonic shock, were decreased by Ca2+ removal; they were both elevated when extracellular [Ca2+] ([Ca2+]o) was raised or following Na+ removal. The hypotonicity-induced rise in [Ca2+]i was inhibited by exposure of cells to gadolinium (Gd3+; 10 micromol/l), an inhibitor of mechanosensitive channels. [Ca2+]i was also elevated following treatment of cells with thapsigargin (10 micromol/l), an inhibitor of the Ca2+ pump of intracellular stores. CONCLUSIONS: A model is presented which interprets these findings in relation to Ca2+ homeostasis in equine articular chondrocytes, including the presence of mechanosensitive channels allowing Ca2+ entry, a Na+/Ca2+ exchanger for removal of intracellular Ca2+ and intracellular stores sensitive to thapsigargin. POTENTIAL RELEVANCE: A more complete understanding of Ca2+ homeostasis in equine chondrocytes may allow development of future therapeutic regimes to ameliorate joint disease.     

Mechanism of carbon monoxide-induced relaxation in the guinea pig ileal smooth muscle

The mechanism of carbon monoxide (CO)-induced relaxation were investigated in the guinea-pig ileum. CO (10%) inhibited the 40 mM KCl-induced contraction. This effect was antagonized by ODQ (1 microM), a soluble guanylate cyclase inhibitor. In contrast, CO did not inhibit the 40 mM KCl-induced increase in cytosolic Ca2+ level ([Ca2+]i). Cumulative addition of KCl induced a graded increase in both [Ca2+]i and muscle tension. In the presence of CO, the increase in muscle tension was attenuated whereas the increase in [Ca2+]i was only slightly decreased. Thus, the [Ca2+]i-tension relationship constructed by cumulative addition of KCl shifted downwards in the presence of CO. Using the patch clamp, CO was found to have little effect on the peak Ba currents (I(Ba)) when voltage was stepped from -60 mV to 0 mV. From these results, we conclude that CO inhibits contraction of guinea-pig ileum mainly by the decrease in the sensitivity of contractile elements to Ca2+ via a cyclic GMP-dependent pathway but not by the inhibition of L-type Ca2+ channel.     

Dependence of superoxide anion production on extracellular and intracellular calcium ions and protein kinase C in PMA-stimulated bovine neutrophils.

The involvement of both intracellular and extracellular calcium, as well as the activation of protein kinase C (PKC), in phorbol myristate acetate (PMA)-stimulated respiratory burst in bovine neutrophils has been studied. PMA significantly stimulated the superoxide anion production by these cells. The increased production of superoxide anion was inhibited by BAPTA/AM, an intracellular calcium ([Ca2+]i) chelator, but not affected by EGTA, an extracellular calcium ([Ca2+]0) chelator. PMA also induced PKC activation, and a PKC inhibitor, calphostin C, blocked the stimulatory effect of PMA on superoxide anion production by the neutrophils. Therefore, we conclude that PMA-induced respiratory burst in bovine neutrophils is [Ca2+]i- but not [Ca2+]0-dependent, and also requires PKC activation.     

The effect of progesterone on oxytocin-stimulated intracellular Ca2+ mobilisation and prostaglandin secretion in porcine endometrium

We have studied in the porcine endometrium the expression of oxytocin receptor (OTR) mRNA and the effect of progesterone (P4) on oxytocin/oxytocin receptor (OT/OTR) function concerning intracellular Ca2+ mobilisation ([Ca2+]i), prostaglandin F2alpha (PGF2alpha) and E2 (PGE2; PG) secretion. Tissue was taken from cyclic and early pregnant pigs (days 14-16). A higher expression of OTR mRNA (P < 0.05) was observed in the endometrium of cyclic than pregnant pigs. The stimulatory (P < 0.05) effect of OT (10(-7) M) on [Ca2+]i mobilisation was noticed within 15-60 s and 30-60 s in endometrial stromal cells of cyclic and pregnant pigs, respectively. In the presence of P4 (10(-5) M) basal and OT-stimulated [Ca2+]i concentrations decreased in stromal cells during luteolysis and pregnancy. In stromal cells P4 delayed mobilisation of [Ca2+]i in response to OT by 15 s during luteolysis and had no effect during pregnancy. In cyclic and pregnant epithelial cells OT stimulated mobilisation of [Ca2+]i in 45 s and 60 s, respectively. Oxytocin increased (P < 0.05) PGF2alpha secretion during luteolysis and pregnancy and PGE2 during luteolysis from endometrial slices. Progesterone did not inhibit this stimulatory effect. During luteolysis OT increased (P < 0.05) PGF2alpha in epithelial and stromal cells and PGE2 secretion in epithelial cells. In the presence of P4 this effect of OT was reduced only in stromal cyclic cells (6 h culture). The presence of P4 decreased the effect of OT on [Ca2+]i mobilisation only in stromal cells. We found that, in most conditions, P4 did not inhibit the OT-stimulated secretion of PG in the porcine endometrium.