NO对家兔Oddi括约肌肌电活动和血压的影响

应用３２只家兔观察一氧氮对Ｏｄｄｉ括约肌肌电和血压的影响。静脉注射ＮＯ合酶抑制剂Ｎ＾Ｇ－硝基－Ｌ－精氨酸,可见ＳＯ肌电振幅增大和血压升高,Ｌ－ＮＮＡ所致的肌电活动增强可Ｌ－精氨酸反转。     

中缝隐核对兔奥迪氏括约肌肌电活动的影响

实验用电生理学和微量注射法观察了兔中缝隐核（NRO）对奥迪氏括约肌肌电活动的影响,动物禁食但自由饭水,18－24h手用乌拉坦（1.0g/kg)静脉麻醉,用双极康铜丝电极引导奥迪氏括约肌肌电,发现NRO内微注射谷氨酸（340mmol/L,0.4ul)可使奥迪氏括约肌肌电活动加强,与在NRO内微量注射生理盐水或者将谷氨酸（340mmol/L,0.4ul)注射到NRO以外的地方相比,具有显著差异（P＜0．01）,NRO 微量注射N-methy-D-aspartate(NM-DA)受体阻断剂氯胺酮（180mmol/L,0.1ul),可消除谷氨酸的效应,而将微量非NMDA受体阻断剂CNQX（2mmol/L,0.1ul)注入NRO,可使奥迪氏括约肌肌电加强,外周应用M－受体阻断剂阿托品（0.2mg/kg)或双侧颈部迷走神经切断,可阻断微量谷氨酸注射到NRO内所引起的效应,静脉注射α－受体阻断剂酚妥拉明（1.5mg/kg),心得安（1.5mg/kg)或自T3－4处切断脊髓,对NRO内微量注射谷氨酸的效应没有影响,结果提示,NRO对奥迪氏括约肌运行有调节作用,其中谷氨酸主要通过NMDA受体兴雷奥迪氏括约肌肌电活动,其传出途径是迷走神经和外周M受体。     

Down Regulated Expression of the β1 Subunit of the Big-conductance Ca^2＋ Sensitive K＋ Channel in Sphincter of Oddi Cells from Rabbits Fed with a High Cholesterol Diet

Hypercholesterolemia,which is closely related to gallbladder bile stasis,can cause sphincter ofOddi dysfunction (SOD) by increasing the tension of sphincter of Oddi (SO).Intracellular calcium ionconcentration ([Ca~(2 )]_i) could influence the tension of SO.The β1 subunit of the big-conductance Ca~(2 )sensitive K~ channel (BK_(Ca) can enhance the sensitivity of the BK_(Ca) channel to [Ca~(2 )]_i.Absence and decline ofthe BK_(Ca) channel subunit β1 could lead to many diseases.However,the relationship betweenhypercholesterolemia and the expression of β1 subunit is not well understood.In this study,we successfullyexpressed and purified the rabbit BK_(Ca) β1 subunit protein and prepared its polyclonal antibody.The specificityof the prepared antibody was determined by western blotting.A SOD rabbit model induced by a high cholesteroldiet was established and the expression of the β1 subunit of SO was determined by immunohistochemicalstaining and western blotting.Compared with the controls,our results demonstrated that hypercholesterolemiacould decrease the expression of the β1 subunit in the SO cells from rabbits.This indicates that lowerexpression of BK_(Ca) channel β1 subunit might induce SOD.     

Nutritional control of thyroid morphogenesis through gastrointestinal hormones

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Action of cholecystokinin on the dog sphincter of Oddi: influence of anti-cholinergic agents.

Effects of cholecystokinin (CCK) on bile flow through the sphincter of Oddi (SO) were studied in anaesthetized dogs. Intravenous injection of CCK (0.25, 0.5, 1 and 2 IDU/Kg) elicited a dose-dependent reduction in flow through the SO in the first minutes after CCK administration. Pirenzepine and atropine decreased significantly (P less than 0.05) by 29% and a 40% respectively the inhibitory effect induced by 1 IDU/Kg of CCK, whereas hexamethonium elicited an increase in the inhibitory effect induced by 0.5 IDU/Kg of CCK (P less than 0.05). Intravenous infusion of cummulative doses of CCK had different effects according to the dose infused. Lower doses (0.025 and 0.05 IDU/Kg/min) increased transphincteric flow, however, high doses (0.1, 0.2 and 0.4 IDU/Kg/min) were inhibitory. These finding indicated that CCK had two effects on the SO : firstly, a contractile effect, probably mediated through a direct myogenic action and neuronal release of ACh, and secondly a relaxant effect, probably mediated by stimulation of inhibitory postganglionic neurons.     

Distribution of opioidergic,sympathetic and neuropeptide Y-positive nerves in the sphincter of Oddi and biliary tree of the monkey,Macaca fascicularis

Summary The opioidergic, sympathetic and neuropeptide Y-positive innervation of the sphincter of Oddi (common bile duct sphincter and pancreatic duct sphincter), as well as other segments of the extrahepatic biliary tree was studied in the monkey by use of immunohistochemistry. Methionine-enkephalin-positive nerves were seen to innervate the smooth muscle of all portions of the sphincter of Oddi and also local ganglion cells. No methionine-enkephalin-positive nerves could be detected in the common bile duct, pancreatic duct or gallbladder. Tyrosine hydroxylase-positive nerves occurred between smooth muscle bundles and also ran to local ganglion cells as well as along the common bile duct. Neuropeptide Y-positive nerves were observed within smooth muscle of the sphincter of Oddi (all portions), common bile duct, pancreatic duct and gallbladder. No evidence of any differential innervation of the pancreatic duct and common bile duct sphincters could be detected with these markers.     

The effect of 1-week feed restriction on performance,digestibility of nutrients and digestive system development in the growing rabbit

A 3 to 4 week feed restriction of about 20% to 25% of the free intake is widely applied in rabbit breeding systems to reduce post-weaning digestive disorders. However, a short intensive feed restriction is described in few studies and can be beneficial for growing rabbits due to a longer re-alimentation period. The aim of this study was to evaluate the effect of ad libitum (AL) and two restriction levels of feeding (50 and 65 g/rabbit per day) applied for 1 week on performance, gastrointestinal morphology and physiological parameters during the restriction and during the re-alimentation period. Rabbits were divided into three experimental groups: AL rabbits were fed AL, R1 rabbits were restricted from 42 to 49 days of age and received 50 g daily (29% of AL) and R2 rabbits were restricted at the same age and were fed 65 g of feed daily (37% of AL). In the 1^st week after weaning and in the weeks after restriction, all the groups were fed AL. During the restriction period, daily weight gain (DWG) in R1 significantly dropped to 11% (experiment 1) and 5% (experiment 2) compared with rabbits in the AL group, although they were fed 29% of AL, whereas in the R2 group it decreased to 20% (experiment 1) and 10% (experiment 2). In the week following feed restriction, DWG in the restricted groups increased (P<0.001) to 166% and 148% in R1 and to 128% and 145% in R2. Restricted rabbits in both the experiments reached up to 90% to 93% of the final live weight (70 days) of the AL group. Over the entire experimental period, feed restriction significantly decreased feed intake to 85% to 88% of the AL group; however, the feed conversion ratio was lower (P<0.05) only in experiment 1 (−6% in R1 and −4% in R2). Digestibilities of CP and fat were not significantly higher during the restriction period and during the 1^st week of re-alimentation compared with the AL group. Significant interactions between feeding regime and age revealed the shortest large intestine in the AL group at 49 days of age and the longest at 70 days in the AL and R1 groups. Small intestinal villi were significantly higher and the crypts were significantly deeper in the restricted groups. It could be concluded that short intensive feed restriction increased digestible area in the small and large intestines, including the height of villi and depth of crypts, which might be involved in the compensatory growth and defence mechanism.     

Differentiation of the effects of antispasmodic agents on the electromyogram of the sphincter of Oddi and the duodenum of the alert rabbit

The electrical activity of the sphincter of Oddi and gastro-intestinal tract had been recorded on 21 negative atropineesterase conscious rabbits by means of chronically implanted electrodes located in the digestive wall. An analysis of the action of different spasmolytic and analgesic drugs was realized. Electromyograms of the sphincter of Oddi presented (a) isolated or in series spike potentials occurring independently of electrical activities of the duodenum (b) recurring spike potentials correlated with intestine spikes. The independent activity of the sphincter of Oddi was not controlled by the cholinergic system, contrary to the intestine-dependent activity (effect of the fempiverinium, atropine like drug). The pitofenone had inhibited the spike potentials of both the sphincter and intestine because its papaverinic effect. The noramidopyrine, analgesic drug without morphine-like effects, had induced activation and inhibition at low and high posology respectively.     

Effect of regulatory polypeptides on the substance P stimulated lower esophageal sphincter pressure in pigs

The effect of graded doses of vasoactive intestinal polypeptide (VIP), enkephalin, neuropeptide Y (NPY), gastrin-17, pentagastrin, cholecystokinin (CCK)-4, CCK-8, neurotensin, somatostatin, and thyrotropin-releasing hormone (TRH) on the substance P (SP)-stimulated lower esophageal sphincter pressure (LESP) in anaesthetized pigs was studied by direct infusion of the peptides into the arterial supply of the lower esophageal sphincter (LES). Infusion of SP in a dose of 20 pmol/kg per min for 3 min significantly increased the LESP (P less than 0.01). Simultaneous VIP infusion at 5--40 pmol/kg per min showed a dose-dependent inhibition of the effect of SP on the LESP. None of the other peptides had any effect on the LESP during simultaneous infusion of SP. Pharmacological blockade by atropine (250 mu/kg) or guanethidine (1 mg/kg) had no effect on the SP-stimulated LESP. In conclusion, the SP-induced stimulation of the LESP is abolished by VIP, and both peptides seem to play a role in the complex regulation of the LESP.     

Uteroglobin in the rabbit

Summary Rabbit uterine uteroglobin (UGL) was studied by electrophoretic and immunological methods following normal copulation, after ovariectomy and progesterone treatment, 17-oestradiol and combined progesterone treatment, 17-oestradiol treatment alone and after HCG-induced pseudopregnancy. Electrophoretic studies show the amount of ULG in uterine secretions, the immunological investigations indicate the intracellular localization of ULG and the distribution of ULG-positive cells in the endometrium.No obvious differences were found between the uteri 7 days after injection with chorion-gonadotropin and those 7 days following normal copulation. No differences could be demonstrated between the uteri of animals 35 days following ovariectomy and subsequent progesterone treatment on Days 31–33 and those of normal 7 d. post coitum (p.c.) animals. Uteri from animals treated with progesterone on Days 2–5 p.c. contained more ULG-positive cells than controls. 17-oestradiol treatment with and without subsequent progesterone treatment resulted, in both gravid and ovariectomized animals, in the formation of a tall columnar endometrial epithelium. Treatment with 17-oestradiol on Days 1 and 2 p.c. led to a decrease in the number of UGL-positive cells at 7 days p.c. Even after ovariectomy with 17-oestradiol substitution, UGL-positive cells were still present in the endometrium. However a secretion of any magnitude could not be detected. The importance of differentiation between synthesis and secretion (= release) as distinct phases of the glandular response is especially emphasised by the latter findings.Supported by Grant No. Ki 154/5-6, Deutsche Forschungsgemeinschaft     

Reevaluation of lipolytic activity of growth hormone in rabbit adipocytes

The lipolytic activities of porcine pituitary fractions and purified growth hormone (GH) from human (h), porcine (p), ovine (o) and rabbit (Rb) origin as well as ovine placental lactogen (oPL), were compared to that of ACTH on rabbit adipocytes. All the GH preparations and oPL were equivalent in inhibiting the binding of labelled oGH to liver plasma membranes from pregnant rabbits. ACTH, and to a lesser extent porcine pituitary fractions and hGH, stimulated free fatty acid production by isolated adipocytes. The sensitivity of the adipocytes to these factors was increased when adenosine deaminase was added to the incubation medium. But, RbGH, pGH, oGH and oPL had no effect. We conclude that GH is not directly involved in the control of lipolysis in rabbit adipocytes and that the effect of hGH is rather due to a contamination of this preparation by other pituitary factors.     

Feed restriction strategy in the growing rabbit. 1. Impact on digestion, rate of passage and microbial activity

The effects of a quantitative feed restriction on the digestive physiology of the young rabbit remain largely unclear. Several digestive functions were thus analysed in the rabbit after weaning, using a monofactorial design that produces a linear reduction of the intake, from ad libitum (AL group) to 80%, 70% and 60% of AL (I80, I70 and I60). The restriction programme was applied by giving a daily meal during 21 days after weaning (34 days), and then a 4-day transition period was managed where the feed intake was fixed at 80% of the AL group, before to be fed ad libitum till 69 days of age. The young rabbit quickly adapted to the restriction programme, since within 4 days after weaning they ate totally their ration within 6-7 h after the feed distribution at 8:00, while AL animals consumed 75% of their feed between 15:00 and 8:00. From 55 to 59 days old, rabbits of I70 and I60 groups reached the intake of the I80 group within 1 day, and then the feed intake of restricted animals increased progressively without over-eating. From 54 to 69 days old, the intake of the four groups did not differ and averaged 143.7 g/day per rabbit. During restriction, the live weight and the weight gain decreased linearly with the restriction level. From 55 to 69 days, the weight gain increased linearly according to the restriction level previously applied, but the final weight of restricted rabbits remained lower than AL ones (-3%, -5% and -7%, respectively, for I80, I70 and I60). After 7 days of restriction, the digestibility was not significantly affected by the restriction level, except for crude protein that presented a slightly higher (+1.5 unit, P = 0.05) coefficient in I70 and I60 groups. The mean retention time (MRT) of particles increased by 50% for restricted animals (mean: 26.2 h for I80 and I60) compared to the AL ones, while that of the liquid phase (three times longer than the particles) was linearly and moderately increased with restriction (+20% between AL and I60). In restricted groups, the caecal pH was lower (-0.3 unit, P < 0.05) and could be related to their higher volatile fatty acid (VFA) concentration (+16 mmol/l compared to AL, P < 0.05). The fermentation pattern, ammonia concentration and the caecal bacterial fibrolytic activity remained similar among treatments, although the butyrate proportion tended to be higher in restricted animals. Impact of feed restriction on performances and digestive health is reported in the second part of this study.     

Comparison of angiotensin II (Ang II) effects in the internal anal sphincter (IAS) and lower esophageal sphincter smooth muscles

Studies were performed to compare the actions of Ang II in the internal anal sphincter (IAS) vs. lower esophageal sphincter (LES) smooth muscles in vitro, in opossum and rabbit. Studies also were carried out in isolated smooth muscle cells. In opossum, Ang II produced no discernible effects in the IAS, but did produce a concentration-dependent contraction in the LES. Conversely, in the rabbit, while Ang II caused a modest response in the LES, it caused a significant contraction in the IAS. The contractile responses of Ang II in the opossum LES were mostly resistant to different neurohumoral antagonists but were antagonized by AT1 antagonist losartan. AT2 antagonist PD 123,319, rather than inhibiting, prolonged the contractile action of Ang II. The contractile actions of Ang II in the opossum LES were not modified by the tyrosine kinase inhibitors (genistein and tyrphostin 1 x 10(-6) M) but were partially attenuated by the PKC inhibitor H-7 (1 x 10(-6) M), Ca2+ channel blocker nicardipine (1 x 10(-5) M), Rho kinase inhibitor HA-1077 (1 x 10(-7) M) or p(44/42) MAP kinase inhibitor PD 98059 (5 x 10(-5) M). The combination of HA-1077 and H-7 did not cause an additive attenuation of Ang II responses. Western blot analyses revealed the presence of both AT1 and AT2 receptors. We conclude that Ang lI-induced contraction of sphincteric smooth muscle occurs primarily by the activation of AT1 receptors at the smooth muscle cells and involves multiple pathways, influx of Ca2+, and PKC, Rho kinase and p(44/42) MAP kinase.     

The thin limbs of Henle's loop in the rabbit

Summary The thin limbs of short and long loops of Henle of the rabbit kidney were studied by freeze fracture techniques. According to TEM studies of thin sections four segments are discernible: descending thin limbs of short loops, descending thin limbs of long loops, subdivided into an upper and a lower part, and ascending thin limbs (Kaissling and Kriz 1979). This division is supported by findings obtained with the freeze fracture technique and based on differences in the organization of the junctional complexes as well as on differences in the internal morphology of the cell membranes. The descending thin limbs of short loops have junctional complexes established by several closely arranged junctional strands and numerous desmosomes. The upper parts of the long descending thin limbs have tight junctions consisting of a variable number of strands; their outstanding characteristic after freeze fracture is a high density of intramembrane particles in both luminal and baso-lateral membranes. The tight junctions of the lower part of the long descending thin limbs consist of several anastomosing junctional strands, which are, in contrast, loosely arranged; the cell membranes contain only a sparse population of intramembrane particles. The ascending thin limbs are characterized by shallow tight junctions (frequently consisting of only one single junctional strand). Moreover, the epithelial cells of this segment are heavily interdigitated; thereby the tight junctions are correspondingly lengthened.In addition, this study presents further evidence that remarkable species differences occur among thin limb epithelia. The junctional complexes of the long descending thin limbs of the rabbit are organized quite differently from those of small rodents (e.g., rat, Psammomys).The data of this study support the concept that the tight junctions are the main determinant of ionic conductances of the paracellular pathway. However, with reference to recent findings from microperfusion studies, it becomes obvious that a correlation of the junctional morphology with the transepithelial water permeability is lacking, at least for the thin limbs.This investigation was supported by the Deutsche Forschungsgemeinschaft; project Kr 546 Henlesche Schleife     

The effect of adipokinetic hormones on the activity of digestive enzymes

The role of the adipokinetic hormone (AKH) in the control of protease, amylase and lipase activities is examined using the cockroach Periplaneta americana and the fruit fly Drosophila melanogaster as model species. The effects of Peram‐CAH‐I and ‐II on the activity of cockroach digestive enzymes in the gastric caeca and midgut are measured both in vivo and in vitro. The results show the activity of proteases, amylases and lipases in both parts of the gut: amylase activity is higher in the gastric caeca than in the midgut; lipase activity presents the opposite trend; and protease activity is similar in both organs. The applied hormones stimulate the activity of all digestive enzymes, although this stimulation is not uniform; AKHs affect enzymes selectively, and in some cases unequally, in the gastric caeca and midgut. No substantial differences between Peram‐CAH‐I and ‐II stimulation are recorded. The in vitro results demonstrate that AKH stimulates digestive enzyme activity directly. In agreement with the cockroach results, enzymatic activity in D. melanogaster larvae producing nonfunctional AKH is lower than that in the larvae with ectopically expressed Akh gene, where enzyme activity reaches or even exceeds that of the controls. Overall, the results demonstrate the active role of AKHs in the stimulation of digestive enzyme activity in insects.     

Expression and alteration of BKCa channels in the sphincter of Oddi's from rabbits with hypercholesterolemia

This study aimed to investigate the expression and function of BK_Ca channels in the Sphincter of Oddi (SO) in a rabbit model of hypercholesterolemia (HC). New Zealand white rabbits were randomly divided into 2 groups: the control group was fed standard chow (n = 18) whereas the high-cholesterol group was fed cholesterol-enriched chow containing 1.5% cholesterol (n = 18). The serum cholesterol level was significantly greater in the HC groups than in the control group, but there was no significant difference in body weight between the control and HC groups. Although the total protein expression of BK_Ca α- and β₁-subunit was not significantly different between the control and HC groups, the Tyr-phosphorylation of BK_Ca α-subunit was significantly decreased in the HC group than in the control group. In addition, hypercholesterolemia significantly increased Acetylcholine (ACh)-induced contraction of the SO rings. Pretreatment with 30 μM NS1619, a BK_Ca channel agonist, significantly reduced ACh-induced contraction of the SO rings in HC rabbits. Moreover, pretreatment with 100 μM Na₃OV₄, a protein tyrosine phosphatase inhibitor, significantly reduced ACh-induced contraction of the SO rings in HC rabbits, whereas it significantly increased upon pretreating with 10 μM Genistein, a tyrosine kinase inhibitor. Whole-cell patch clamp recordings showed that BK_Ca current density was significantly lower in SOSMCs from HC group than that from control group. Our findings suggest that hypercholesterolemia-induced downregulation of BK_Ca channel, and Tyr-phosphorylation of BK_Ca α-subunit may contribute to the hyperresponsiveness of the SO ring in HC rabbits.     

Activation of phospholipase D by endothelin-1 and other pharmacological agents in rabbit iris sphincter smooth muscle

The stimulation of phospholipase D (PLD) activity by endothelin-1 (ET1) was investigated in rabbit iris sphincter perlabelled with [³H]myristic acid. In the presence of 0.5% ethanol, ET1 caused a time- and dose-dependent increase in the production of [³H]phophatidylethanol ([³H]PEt). Within 30 s the peptide increased PEt formation by 30% and after 5 min increased it by 140%. The ₅₀ value for ET1-stimulated PEt formation was found to be 30 nM. This value is appreciably lower than the ₅₀ we previously obtained for ET1-induced inositol triphosphate production (45 nM), but considerably higher than that for arachidonic acid release (1 nM). PEt formation was significantly stimulated by prostaglandin F₂₀, phorbol 12,13-dibutyrate (PDBu), chloroform, A23187 and A1F₄⁻, but it was not affected by carbachol or the platelet-activating factor. PDBu-stimulated PEt formation was blocked by staurosporine and it was not potentiated by A23187. Staurosporine had no effect on ET1-stimulated PEt formation. Our data indicate that ET1 stimulation of PLD occurs independently of protein kinase C activation, phospholipase C activation and intracellular Ca²⁺ mobilization, and phospholipase A₂ activation. In this tissue the ET1 receptor is probably coupled to the three phospholipases through several G-proteins, and this appears to be species and receptor type specific.