Protease activated receptor 2 and epidermal growth factor receptor are involved in the regulation of human sperm motility

Aim： To investigate mechanisms of tryptase-induced reduction of sperm motility and explore whether epidermal growth factor receptor （EGF-R） and protease activated receptor 2 （PAR-2）- associated pathways are involved. Methods： Fresh semen was collected from healthy donors （n = 15）. Semen parameters and quality were assessed in accordance with the World Health Organization （WHO） criteria. Swim-up sperm were fixed and subjected to immunocytochemistry and immunoelectronmicroscopy with specific antibodies directed against PAR-2 and EGF-R. Protein extractions from swim-up spermatozoa were analyzed by Western blotting with antibodies for both receptors. Motility of spermatozoa was evaluated by computer-assisted semen analysis. Results： Immunocytochemistry found PAR-2 and EGF-R in approximately 30% of examined human ejaculated spermatozoa. Both receptors were localized in the plasma membrane. Like tryptase, the PAR-2 synthetic agonist SLIGKV reduced sperm motility, and this effect was inhibited by application of two specific EGF-R pathway blockers （AG1478 and PD168393）. Conclusion： The observed reduction of sperm motility by tryptase through the PAR-2 receptor involves EGF-R pathways.

Effect of exogenous hydrogen sulfide in the nucleus tractus solitarius on gastric motility in rats

BACKGROUND Hydrogen sulfide(H2S)is a recently discovered gaseous neurotransmitter in the nervous and gastrointestinal systems.It exerts its effects through multiple signaling pathways,impacting various physiological activities.The nucleus tractus solitarius(NTS),a vital nucleus involved in visceral sensation,was investigated in this study to understand the role of H2S in regulating gastric function in rats.AIM To examine whether H2S affects the nuclear factor kappa-B(NF-κB)and transient receptor potential vanilloid 1 pathways and the neurokinin 1(NK1)receptor in the NTS.METHODS Immunohistochemical and fluorescent double-labeling techniques were employed to identify cystathionine beta-synthase(CBS)and c-Fos co-expressed positive neurons in the NTS during rat stress.Gastric motility curves were recorded by inserting a pressure-sensing balloon into the pylorus through the stomach fundus.Changes in gastric motility were observed before and after injecting different doses of NaHS(4 nmol and 8 nmol),physiological saline,Capsazepine(4 nmol)+NaHS(4 nmol),pyrrolidine dithiocarbamate(PDTC,4 nmol)+NaHS(4 nmol),and L703606(4 nmol)+NaHS(4 nmol).RESULTS We identified a significant increase in the co-expression of c-Fos and CBS positive neurons in the NTS after 1 h and 3 h of restraint water-immersion stress compared to the expressions observed in the control group.Intra-NTS injection of NaHS at different doses significantly inhibited gastric motility in rats(P<0.01).However,injection of saline,first injection NF-κB inhibitor PDTC or transient receptor potential vanilloid 1(TRPV1)antagonist Capsazepine or NK1 receptor blockers L703606 and then injection NaHS did not produce significant changes(P>0.05).CONCLUSION NTS contains neurons co-expressing CBS and c-Fos,and the injection of NaHS into the NTS can suppress gastric motility in rats.This effect may be mediated by activating TRPV1 and NK1 receptors via the NF-κB channel.

Oxytocin decreases colonic motility of cold water stressed rats via oxytocin receptors

AIM: To investigate whether cold water intake into the stomach affects colonic motility and the involvement of the oxytocin-oxytocin receptor pathway in rats. METHODS: Female Sprague Dawley rats were used and some of them were ovariectomized. The rats were subjected to gastric instillation with cold (0-4 degrees C, cold group) or room temperature (20-25 degrees C, control group) saline for 14 consecutive days. Colon transit was determined with a bead inserted into the colon. Colonic longitudinal muscle strips were prepared to investigate the response to oxytocin in vitro. Plasma concentration of oxytocin was detected by ELISA. Oxytocin receptor expression was investigated by Western blot analysis. Immunohistochemistry was used to locate oxytocin receptors. RESULTS: Colon transit was slower in the cold group than in the control group (P < 0.05). Colonic smooth muscle contractile response to oxytocin decreased, and the inhibitory effect of oxytocin on muscle contractility was enhanced by cold water intake (0.69 +/- 0.08 vs 0.88 +/- 0.16, P < 0.05). Atosiban and tetrodotoxin inhibited the effect of oxytocin on colonic motility. Oxytocin receptors were located in the myenteric plexus, and their expression was up-regulated in the cold group (P < 0.05). Cold water intake increased blood concentration of oxytocin, but this effect was attenuated in ovariectomized rats (286.99 +/- 83.72 pg/mL vs 100.56 +/- 92.71 pg/mL, P < 0.05). However, in ovariectomized rats, estradiol treatment increased blood oxytocin, and the response of colonic muscle strips to oxytocin was attenuated. CONCLUSION: Cold water intake inhibits colonic motility partially through oxytocin-oxytocin receptor signaling in the myenteric nervous system pathway, which is estrogen dependent. (C) 2014 Baishideng Publishing Group Inc. All rights reserved.

Expression Imbalance of Cholinergic M₂and M₃Receptors Contributes to the Motility Reduction of the Small Intestine in Spleen Qi Deficiency

Objective: To study roles of cholinergic M2 and M3 receptors in the motility reduction of small intestine (SI) in spleen qi deficiency. Methods: 16 male SD rats were randomly divided in the control group and spleen qi deficiency group (model group)—8 rats each group;spleen qi deficiency model of the improper diet and overfatigue was established;the SI propelling rate (SIPR) was used to evaluate the SI motility;ELISA was used to measure concentrations of acetylcholine (ACh), cyclic adenosine monophosphate (cAMP) and protein kinase A (PKA) in the SI tissue;immohistochemistry was employed to detect expressions of cholinergic M2 and M3 receptors. Results: Compared with those in the control group, SIPR was reduced;expression of M2 receptors was increased;and expression of M3 receptors and concentrations of cAMP and PKA were decreased, significantly, in the model group. Conclusions: Expression imbalance of cholinergic M2 and M3 receptors might contribute to the motility reduction of the SI in spleen qi deficiency.

Involvement of Ca^(2+)-activated K^+ Channels in Receptor-Regulated Sperm Motility in Rats

Previous voltage clamp studies have demonstrated the modulation of sperm Ca 2+ activated K + (KCa) channels expressed in Xenopus oocytes by angiotensin II (Ang II) and extracellular ATP via AT 1 receptor and P 2U receptor, respectively. In the present study, we investigated the involvement of KCa channels in receptor regulated sperm motility of the rat using a computer aided sperm analysis system, HTM IVOS, in conjunction with Ca 2+ mobilizing agents, receptor agonists/antagonists and KCa channels blockers. The percentage of motile sperm was increased by ionomycin (0.5 μmol/L), which could be inhibited by K + channel blockers, tetraethylammonium (TEA 1 μmol/L ) or charybdotoxin (ChTX, 300 nmol/L) indicating the presence of KCa channels. Ang II, at low concentration, 10 nmol/L, was found to increase motility, however, at higher concentration, 1 μmol/L, percentage of motility was found to be suppressed. Both stimulatory and inhibitory effects of Ang II could be reversed by losartan, a specific antagonist of AT 1 receptors, but not AT 2 antagonist PD123177, indicating the involvement of AT 1 but not AT2 receptor in mediating both effects. ChTX also abolished both stimulatory and inhibitory effects of Ang II, suggesting the involvement of KCa channels. The percentage of motility was also enhanced by extracellular ATP, a factor known to be involved in sperm activation. The ATP enhanced sperm motility was mimicked by UTP, and inhibited by ChTX and reactive blue, an antagonist of P 2 receptor, indicating the involvement of both P 2U and KCa channels. RT PCR study was also conducted to confirm the expression of KCa channels, AT 1 receptors and P 2U receptor, but not AT 2 receptor, in rat caudal epididymal sperm. The present findings suggest an important role of KCa channels in the regulation of sperm motility by AT 1 and P 2U receptors.

Differential expression of autocrine motility factor receptor (AMFR) mRNA in normal and cancer cells detected by in situ hybridization

The receptor for autocrine motility factor (AMFR) is known to be involved in the process of AMF-mediated cell migration and metastasis. This paper describes the procedures of non-radioactive in situ hybridization (ISH) detection of AMFR mRNA in both paraffin-embedded surgical sections and cultured cells using either biotinylated oligonucleotide probes or digoxigenin-labeled RNA probes. The results showed that the AMFR mRNA was expressed at an enhanced level in hyperplaJstic and malignant tissues of breast and prostate cancer patient surgical specimens, indicating that the elevated AMFR expression was associated with the tissue malignancy Moreover, AMFR mRNA was detected in both normal and earcinoma cells when cultured at a subconfluent density. However, AMFR expression was inhibited in confluent normal (3T3-A31 murine fibroblast and FHs738BL human bladder) cells while it continued to express in carcinoma (J82 human bladder)and metastatic (3T3-M murine fibroblast) cells irrespective of cell density This suggested a cell-cell contact downregulation of AMFR mRNA expression in normal but not in cancer cells. The ISH data obtained in this study are closely consistent with the AMFR protein expression pattern previously reported, implying that the differential expression of AMFR gene may be regulated and controlled at the transcriptional level.

Electroacupuncture at ST25 inhibits jejunal motility:Role ofsympathetic pathways and TRPV1

AIM: To investigate whether electroacupuncture(EA) at ST25 affects jejunal motility in vivo and if so, whether a sympathetic pathway is involved.METHODS: Jejunal motility was assessed using a manometric balloon placed in the jejunum approximately about 3-5 cm away from the suspensory ligament of the duodenum in anesthetized animals. The effects of EA at ST25 were measured in male Sprague-Dawley rats, some of which were treated with propranolol or clenbuterol(EA intensities: 1, 3, 5, 7, and 9 m A), and in male transient receptor potential vanilloid-1(TRPV1)(capsaicin receptor) knockout mice(EA intensities: 1, 2, and 4 m A).RESULTS: Anesthetized rats exhibited three types of fasting jejunal motor patterns(types A, B, and C), and only type C rats responded to EA stimulation. In type C rats, EA at ST25 significantly suppressed the motor activity of the jejunum in an intensity-dependent manner. The inhibitory effect of EA was weakened by propranolol(β adrenoceptor antagonist) and disappeared with clenbuterol(β adrenoceptor agonist) induced inhibition of motility, suggesting that the effect of EA on motility is mediated via a sympathetic pathway. Compared with wild-type mice, EA at ST25 was less effective in TRPV1 knockout mice, suggesting that this multi-modal sensor channel participates in the mechanism. CONCLUSION: EA at ST25 was found to inhibit jejunal motility in an intensity-dependent manner, via a mechanism in which sympathetic nerves and TRPV1 receptors play an important role.

Acupuncture at heterotopic acupoints enhances jejunal motility in constipated and diarrheic rats

AIM: To investigate the effect and mechanism of acupuncture at heterotopic acupoints on jejunal motility, particularly in pathological conditions.

Evaluation of the gastrointestinal anti-motility effect of Anacardium occidentale stem bark extract:A mechanistic study of antidiarrheal activity

Diarrhea is a prevalent gastrointestinal problem associated with fatal implications.It is a huge public health concern that requires better alternatives to current drugs.This study investigated the mechanisms involved in the antidiarrheal activity of Anacardium occidentale(Ao) stem bark extract,a plant commonly used in the management of diarrhea in Nigeria.Methanolic stem bark extract of the plant was partitioned into three fractions:hexane fraction,ethyl acetate fraction(Ao EF) and methanol fraction.In vitro studies on the effect of these fractions on guinea pig ileum(GPI) strips,as well as the modulatory effect of Ao EF on standard agonists-and antagonists-induced GPI contraction and relaxation,revealed Ao EF as the most active fraction.In vivo studies to assess the effect of Ao EF on the dopaminergic,muscarinic,and serotonergic pathways were carried out using gastric emptying(GE) and gastrointestinal transit(GT) as experimental end points.Ao EF was subjected to GC-MS analysis,while the identified compounds were docked with the muscarinic acetylcholine receptor M3(CHRM3) using Autodock Vina.Results indicated that Ao EF inhibited GE and GT via inhibition of CHRM3.In addition,GC-MS analysis revealed the presence of 24 compounds in Ao EF,while docking indicated that octadecanoic acid 2-(2-hydroxylethoxy)ethyl ester exhibited the highest binding affinity to CHRM3.This study indicated that the antidiarrheal activity of Ao is through its antimotility effect via the inhibition of the muscarinic pathway.And since none of the identified compounds exhibited higher binding affinity to CHRM3 relative to loperamide,the antimotility activity of these phytoconstituents may be via synergism.

毛建红茶对大鼠胃肠运动的作用机制研究

为探究毛建红茶(MJBT)治疗“脘腹胀满”的作用机制及活性成分,本研究将正常大鼠随机分为空白组(生理盐水)、阳性对照组(莫沙必利,1 mg·kg^(-1)),以及MJBT水提物(MJBT_HE)高(68 mg·mL^(-1))、中(34 mg·mL^(-1))、低(17 mg·mL^(-1))剂量共计5组进行给药试验,检测MJBT-HE对胃内排空率和小肠推进率的影响;用双抗体夹心酶联免疫吸附试验(ELISA)测定胃泌素(GAS)和胃动素(MTL)两种激素分泌水平变化,并结合胃肠离体肌条运动试验,确定MJBT潜在作用部位和相关受体;经高分辨液质联用仪(LC-HRMS)和高效液相色谱(HPLC)分析MJBT_HE中的主要成分。结果表明,低剂量和中剂量MJBT_HE能显著促进胃肠运动(P<0.05),该作用与GAS和MTL分泌量提升(P<0.05),以及胃、小肠前端收缩张力和频率加强(P<0.05)有关;MJBT_HE可能通过作用于M受体和α受体起效;MJBT主要活性成分为圣草酚-7-O-葡萄糖苷、木犀草素-7-O-葡萄糖苷、圣草酚、木犀草素、柚皮素和咖啡因等物质。本研究为MJBT在消化不良治疗过程中的应用奠定了基础。

弱精子症患者精子锌稳态蛋白、GPR39和ANO1 mRNA的表达变化及其临床意义

目的:探究锌稳态相关蛋白、G蛋白偶联受体39(GPR39)及ANO1 mRNA在弱精子症精子中的表达变化,并分析其与精子运动能力的相关性。方法:选取2022年6月至2023年4月我中心收集的弱精子症患者精液标本(PR+NP<40%,PR<32%,精子浓度>15×10^(6)/ml)40例,正常精液标本(PR+NP≥40%,PR≥32%,精子浓度>15×10^(6)/ml)42例。通过CASA检测精液常规参数及精子活力,测量两组精浆锌的含量,运用实时荧光定量PCR(RT-qPCR)对两组精子锌转运蛋白(ZIP13、ZIP8、ZNT10)、金属硫蛋白(MT1G、MT1、MTF)、GPR39和钙依赖性氯离子通道蛋白(ANO1)表达进行定量检测;运用激光共聚焦检测精子中的游离锌分布;运用免疫荧光染色观察精子中GPR39、MT1蛋白的表达;进一步采用Spearman秩相关分析其与精液参数的相关性。结果:与正常组相比,弱精子症组精浆锌的浓度无明显差异(P>0.05),弱精子症组精子游离锌水平明显降低(P<0.05);RT-qPCR检测结果显示,弱精子症组精子MT1G、MTF、ZIP13、GPR39、ANO1 mRNA相对表达量低于正常组(P<0.05);两组间ZIP8、ZNT10、MT1 mRNA相对表达量无显著性差异(P>0.05);免疫荧光结果显示弱精子组GPR39蛋白表达较低(P<0.05);相关性分析结果显示MT1G、MTF、GPR39、ANO1 mRNA相对表达量与前向运动精子百分率和精子活动率呈正相关(P<0.05)。结论:弱精子症患者精子锌稳态蛋白(MT1G、MTF、ZIP13)、GPR39和ANO1 mRNA表达下调且其表达与精子运动能力呈正相关。

多不饱和脂肪酸对Δ15 Des转基因小鼠睾丸结构和功能的影响及机制

目的探讨多不饱和脂肪酸(PUFAs)对Δ15脂肪酸去饱和酶(Δ15 Des)转基因小鼠睾丸结构和功能的影响及机制。方法取C57BL/6野生型(WT)雄性小鼠和Δ15 Des转基因雄性小鼠分别作为对照组(n=5,WT组)和实验组(n=5,TG组),均饲喂含6%花生四烯酸(ARA)饲料8周,麻醉处死,分离睾丸组织及附睾组织,采用气相色谱(GC)检测睾丸组织中脂肪酸的组成及含量,采用精子质量分析仪(CASA)检测附睾组织中精子形态及运动活力[精子总活力、直线运动精子活力、精子平均路径速度(VAP)、精子曲线速度(VCL)及精子直线速度(VSL)],采用苏木精-伊红(HE)染色观察睾丸组织的形态学特征,采用实时荧光定量PCR(RT-qPCR)检测2组小鼠睾丸组织中游离脂肪酸受体1(FFAR1)、FFAR2、FFAR3、FFAR4及过氧化物酶体增殖物激活受体α(PPARα)、PPARβ、PPARγ信使RNA(mRNA)的表达,采用蛋白质印迹法检测2组小鼠睾丸组织中FFAR4和PPARγ蛋白的水平。结果与WT组相比,TG组小鼠睾丸组织中ARA与二十二碳四烯酸(DTA)含量降低(P<0.05或P<0.01),亚油酸(LA)、二十碳五烯酸(EPA)、二十二碳六烯酸(DHA)含量升高(P<0.05或P<0.01);2组小鼠的精子形态及数量无明显差异,TG组小鼠精子的直线运动精子活力、VSL较WT组升高(P<0.05),精子总活力、VAP、VCL无差异(P>0.05);与WT组相比,TG组小鼠睾丸组织曲细精管内空泡较少,成熟精子增多;与WT组相比,TG组小鼠睾丸组织中FFAR4、PPARαmRNA表达上调(P<0.01或P<0.05),PPARβmRNA表达下调(P<0.05),FFAR4蛋白水平增加(P<0.05)。结论n-3 PUFAs可改善Δ15 Des转基因小鼠睾丸组织的结构和功能,其机制可能与结合FFAR4、激活下游信号分子有关。

Effects of 5-HT2B, 5-HT3 and 5-HT4 receptor antagonists on gastrointestinal motor activity in dogs

AIM:To study the effects of 5-hydroxytryptamine(5-HT)receptor antagonists on normal colonic motor activity in conscious dogs.METHODS:Colonic motor activity was recorded using a strain gauge force transducer in 5 dogs before and after 5-HT2B,5-HT3 and 5-HT4 receptor antagonist administration.The force transducers were implanted on the serosal surfaces of the gastric antrum,terminal ileum,ileocecal sphincter and colon.Test materials or vehicle alone was administered as an intravenous bolus injection during a quiescent period of the whole colon in the interdigestive state.The effects of these receptor antagonists on normal gastrointestinal motor activity were analyzed.RESULTS:5-HT2B,5-HT3 and 5-HT4 receptor antagonists had no contractile effect on the fasting canine terminal ileum.The 5-HT3 and 5-HT4 receptor antagonists inhibited phaseⅢof the interdigestive motor complex of the antrum and significantly inhibited colonic motor activity.In the proximal colon,the inhibitory effect was dose dependent.Dose dependency,however,was not observed in the distal colon.The 5-HT2B receptor antagonist had no contractile effect on normal colonic motor activity.CONCLUSION:The 5-HT3 and 5-HT4 receptor antagonists inhibited normal colonic motor activity.The5-HT2B receptor antagonist had no contractile effect on normal colonic motor activity.

Gallbladder motor function, plasma cholecystokinin and cholecystokinin receptor of gallbladder in cholesterol stone patients

AIM: To study the interactive relationship of gallbladder motor function, plasma cholecystokinin (CCK) and cholecystokinin A receptor (CCK-R) of gallbladder in patients with cholesterol stone disease.METHODS: Gallbladder motility was studied by ultrasonography in 33 patients with gallbladder stone and 10 health subjects as controls. Plasma CCK concentration was measured by radioimmunoassay in fasting status (CCK-f) and in 30 min after lipid test meal (CCK-30).Radioligand method was employed to analyze the amount and activity of CCK-R from 33 gallstone patients having cholecystectomy and 8 persons without gallstone died of severe trauma as controls.RESULTS: The percentage of cholesterol in the gallstone composition was more than 70%. The cholesterol stone type was indicated for the patients with gallbladder stone in this study. Based on the criterion of gallbladder residual fraction of the control group, 33 gallstone patients were divided into two subgroups, contractor group (14 cases)and non-contractor group (19 cases), The concentration of CCK-30 was significantly higher in non-contractor group than that in both contractor group and control group (55.86±3.86 pmol/l vs 37.85±0.88 pmol/l and 37.95±0.74 pmol/L, P＜0.01), but there was no difference between contractor group and control group. Meanwhile no significant difference of the concentration of CCK-f could be observed among three groups. The amount of CCK-R was lower in non-contractor group than those in both control group and contractor group (10.27±0.94 fmol/mg vs24.59±2.39 fmol/mg and 22.66±0.55 fmol/mg, P＜0.01).The activity of CCK-R shown as KD in non-contractor group decreased compared to that in control group and contractor group. Only was the activity of CCK-R lower in contractor group than that in control group. The ejection fraction correlated closely with the amount of CCK-R (r = 0.9683,P＜0.01), and the concentration of CCK-30 correlated negatively with the amount of CCK-R closely (r = -0.9627,P＜0.01).CONCLUSION: The distinctive interactive relationship of gallbladder emptying, plasma CCK and CCK-R in gallbladder from this study suggested that the defect of CCK-R may be a key point leading to the impairment of gallbladder motor function and the pathogenesis of cholesterol gallstoneformation may differ in two subgroups of gallstone patient,gallbladder non-contractor group or contractor group.

G protein-coupled estrogen receptor in colon function, immune regulation and carcinogenesis

Estrogens play important roles in the development and progression of multiple tumor types.Accumulating evidence points to the significance of estrogen action not only in tumors of hormonally regulated tissues such as the breast,endometrium and ovary,but also in the development of colorectal cancer(CRC).The effects of estrogens in physiological and pathophysiological conditions are mediated by the nuclear estrogen receptorsαandβ,as well as the membranebound G protein-coupled estrogen receptor(GPER).The roles of GPER in CRC development and progression,however,remain poorly understood.Studies on the functions of GPER in the colon have shown that this estrogen receptor regulates colonic motility as well as immune responses in CRC-associated diseases,such as Crohn’s disease and ulcerative colitis.GPER is also involved in cell cycle regulation,endoplasmic reticulum stress,proliferation,apoptosis,vascularization,cell migration,and the regulation of fatty acid and estrogen metabolism in CRC cells.Thus,multiple lines of evidence suggest that GPER may play an important role in colorectal carcinogenesis.In this review,we present the current state of knowledge regarding the contribution of GPER to colon function and CRC.

Deficiency of platelet-derived growth factor receptor-α-positive cells in Hirschsprung's disease colon

AIM: To investigate whether the expression of platelet-derived growth factor receptor-α-positive (PDGFRα+)-cells is altered in Hirschsprung’s disease (HD).METHODS: HD tissue specimens (n = 10) were collected at the time of pull-through surgery, while colonic control samples were obtained at the time of colostomy closure in patients with imperforate anus (n = 10). Immunolabelling of PDGFRα+-cells was visualized using confocal microscopy to assess the distribution of these cells, while Western blot analysis was undertaken to quantify PDGFRα protein expression.RESULTS: Confocal microscopy revealed PDGFRα+-cells within the mucosa, myenteric plexus and smooth muscle in normal controls, with a marked reduction in PDGFRα+-cells in the HD specimens. Western blotting revealed high levels of PDGFRα protein expression in normal controls, while there was a striking decrease in PDGFRα protein expression in the HD colon.CONCLUSION: These findings suggest that the altered distribution of PDGFRα+-cells in both the aganglionic and ganglionic HD bowel may contribute to the motility dysfunction in HD.

YFa and analogs:Investigation of opioid receptors in smooth muscle contraction

AIM:To study the pharmacological profile and inhibition of smooth muscle contraction by YFa and its analogs in conjunction with their receptor selectivity. METHODS:The effects of YFa and its analogs (D-Ala2) YFa, Y (D-Ala2) GFMKKKFMRF amide and Des-Phe-YGGFMKKKFMR amide in guinea pig ileum (GPI) and mouse vas deferens (MVD) motility were studied using an isolated tissue organ bath system, and morphine and DynA (1-13) served as controls. Acetylcholine was used for muscle stimulation. The observations were validated by specific antagonist pretreatment experiments using naloxonazine, naltrindole and norbinaltor-phimine norBNI. RESULTS:YFa did not demonstrate significant inhibition of GPI muscle contraction as compared with mor-phine (15% vs 62%, P = 0.0002), but moderate inhibition of MVD muscle contraction, indicating the role of κ opioid receptors in the contraction. A moderate inhibition of GPI muscles by (Des-Phe) YFa revealed the role of anti-opiate receptors in the smooth muscle contraction. (D-Ala-2) YFa showed significant inhibition of smooth muscle contraction, indicating the involvement of mainly δ receptors in MVD contraction. These results were supported by specific antagonist pretreatment assays. CONCLUSION:YFa revealed its side-effect-free analgesic properties with regard to arrest of gastroin-testinal transit. The study provides evidences for the involvement of κ and anti-opioid receptors in smooth muscle contraction.

菊苣、陈皮、鸡内金、山楂和余甘子对胃动力障碍大鼠的改善作用研究

该文采用大鼠胃动力障碍模型,研究了菊苣、陈皮、鸡内金、山楂和余甘子等5种药食同源食物的促胃动力效果,主要测定了胃排空率、胃酸、蛋白酶、胃液、血清饥饿素、胃泌素、酪氨基酶受体基因(c-kit)和干细胞因子的转录水平等指标,观察胃窦组织切片。结果发现菊苣、山楂和余甘子能显著提高胃排空速率(P<0.05),菊苣和陈皮显著降低胃游离酸的浓度和排出速率(P<0.05),山楂显著提高胃蛋白酶活性和排出量(P<0.05),鸡内金显著提高了胃蛋白酶排出速率和胃液量(P<0.05)。另外,菊苣、山楂显著提高血清饥饿素含量(P<0.05),余甘子显著提高血清胃泌素含量(P<0.05),菊苣对胃底腺层有修复作用并能显著上调c-kit的转录水平。综上,菊苣、山楂和余甘子对胃动力的促进效果较好,具有开发成促消化食品的潜力。

基于自主神经系统研究厚朴三物汤调控胃肠动力的作用机制

目的基于自主神经系统研究胃肠动力障碍性疾病(DGIMD)可能的发病机制及厚朴三物汤调控胃肠动力的作用机制。方法将40只雄性SD大鼠按计算机数字随机法分为对照组、模型组、厚朴三物汤组、α受体抑制剂组、β受体抑制剂组,每组8只。除对照组外,其余组均腹腔注射20%左旋精氨酸溶液5 d建立DGIMD模型,对照组注射等体积生理盐水。第6天开始,厚朴三物汤组给予厚朴三物汤4.725 g(生药)/(kg·d)灌胃,α受体抑制剂组给予酚苄明2.1 mg/(kg·d)灌胃[剂量逐渐增至4.2 mg/(kg·d)],β受体抑制剂组给予普萘洛尔2.1 mg/(kg·d)灌胃[剂量逐渐增至8.4 mg/(kg·d)],对照组与模型组给予等体积生理盐水灌胃,均每日2次,灌胃7 d。灌胃结束后,进行小肠推进率和胃排空率的测定,HE染色观察胃、空肠、结肠组织病理形态,ELISA法检测血及胃、空肠、结肠组织中去甲肾上腺素(NE)含量,RT-PCR法及Western blot法检测胃、空肠、结肠组织中去甲肾上腺素转运体(NET)mRNA及蛋白表达情况,免疫组织化学法检测胃、空肠、结肠组织中α_(1)、α_(2)、β_(1)、β_(2)型肾上腺素能受体阳性表达情况。结果模型组大鼠的胃排空率、小肠推进率均明显低于对照组(P均<0.05);厚朴三物汤组及α受体抑制剂组小肠推进率与胃排空率均明显高于模型组(P均<0.05)。各组大鼠胃、空肠、结肠组织解剖结构、腺体及肠绒毛完整,未见炎性细胞浸润。模型组大鼠血及胃、空肠、结肠组织中NE含量均明显高于对照组(P均<0.05),各药物组大鼠血及胃、空肠、结肠组织中NE含量均明显低于模型组(P均<0.05),但厚朴三物汤组大鼠血及胃、空肠、结肠组织中NE含量均明显高于α受体抑制剂组和β受体抑制剂组(P均<0.05)。模型组大鼠胃、空肠、结肠组织中NET mRNA表达量均明显低于对照组(P均<0.05),厚朴三物汤组、α受体抑制剂组大鼠胃、空肠、结肠组织中NET mRNA表达量均明显高于模型组(P均<0.05)。模型组大鼠胃、结肠组织中NET蛋白相对表达量明显高于对照组(P<0.05),空肠组织中NET蛋白相对表达量明显低于对照组(P<0.05);各药物组大鼠胃、结肠组织中NET蛋白相对表达量均明显低于模型组(P均<0.05),空肠组织中NET蛋白相对表达量均明显高于模型组(P均<0.05);厚朴三物汤组大鼠胃组织中NET蛋白相对表达量均明显高于α受体抑制剂组和β受体抑制剂组(P均<0.05),结肠和空肠组织中NET蛋白相对表达量均明显低于α受体抑制剂组和β受体抑制剂组(P均<0.05)。各组大鼠胃组织中α_(1)受体蛋白阳性表达平均光密度值比较差异均无统计学意义(P均>0.05)。与对照组比较,模型组大鼠结肠组织中α_(1)、α_(2)、β_(1)受体蛋白阳性表达平均光密度值,胃组织中α_(2)、β_(1)、β_(2)受体蛋白阳性表达平均光密度值,空肠组织中β_(2)受体蛋白阳性表达平均光密度值均明显增高(P均<0.05);空肠组织中α_(1)、α_(2)、β_(1)受体蛋白阳性表达平均光密度值均明显降低(P均<0.05)。与模型组比较,厚朴三物汤组大鼠结肠组织中α_(1)受体蛋白和胃组织中α_(2)、β_(2)受体蛋白阳性表达平均光密度值均明显降低(P均<0.05),空肠组织中α_(1)、β_(1)、β_(2)受体蛋白和结肠组织中β_(2)受体蛋白阳性表达平均光密度值均明显增高(P均<0.05)。结论NET蛋白和α_(2)、β_(1)、β_(2)受体蛋白高表达及NE在胃肠道局部组织与外周血的高含量是导致胃肠动力障碍的重要原因。厚朴三物汤在胃与结肠局部可能有着类似于α_(2)受体抑制剂的效果,其可能通过降低NET蛋白和α_(2)、β_(1)、β_(2)受体蛋白表达及胃肠道局部组织与外周血中NE的含量而发挥治疗DGIMD的作用。

广陈皮对功能性消化不良大鼠胃肠动力的影响

目的探讨广陈皮对功能性消化不良(FD)大鼠胃肠动力的作用及其机制。方法无特定病原体(SPF)级大鼠42只,随机分为正常对照组(7只)和模型组(35只)。采用郭氏夹尾法、不规则喂养和0℃、0.9%氯化钠溶液灌胃等多因素应激干预法制造FD大鼠模型。大鼠造模成功后根据体质量和行为学得分随机分为模型对照组、多潘立酮组、新皮组、十年皮组和模拟陈化组,每组7只,各组大鼠灌胃给药7 d。观察记录大鼠的一般状态和体质量;采用旷场实验观察大鼠的自主活动能力;采用胃肠运动实验观测大鼠的胃残留率和小肠推进率;苏木精-伊红染色后观察大鼠胃窦组织病理形态;采用酶联免疫法测定血清中胃动素和5-羟色胺(5-HT)的含量;采用蛋白质印迹法测定十二指肠中5-HT4R蛋白表达。结果与模型对照组比较,各陈皮给药组一般状态明显改善,体质量和行为学得分明显提高,胃残留率显著降低,小肠推进率显著提高,血清中胃动素和5-羟色胺含量均明显升高,十二指肠5-HT4R蛋白表达明显上调。结论广陈皮可促进FD大鼠的胃肠动力,作用机制可能与上调胃动素和5-HT、5-HT4R有关。广陈皮的陈化时间对FD大鼠胃肠动力作用的影响不明显。