Biomedical subjects

J Y Xu

Publications and source records attributed to J Y Xu.

At least 19 recordsLinked to original sources

PubMed2026

ARID1A Mediates ROS-Induced Osteoclast Activation in TMJ Osteoarthritis.

Excessive osteoclast activation drives rapid subchondral bone destruction, serving as a critical early-stage event precipitating temporomandibular joint osteoarthritis (TMJ-OA). Although epigenetic remodeling is widely recognized as an important interface between pathological environmental signals and genomic response, the specific epigenetic mechanisms translating TMJ-OA-associated stimulation into pathological osteoclast activation remain to be elucidated. Here, using a mechanically induced TMJ-OA mouse model, we identify aberrant reactive oxygen species (ROS) accumulation as a critical upstream driver initiating excessive osteoclast activation and subsequent joint deterioration. By integrating transcriptomic and epigenomic analyses, we delineate the chromatin remodeler AT-rich interaction domain 1A (ARID1A) as an essential oxidative stress sensor within the osteoclast lineage. Mechanistically, ROS accumulation induces ARID1A upregulation and recruitment to the Src enhancer, transcriptionally activating Src and amplifying PI3K-AKT signaling to drive pathological osteoclastogenesis. Conditional knockout of Arid1a in myeloid cells effectively abrogates subchondral bone loss and cartilage destruction in TMJ-OA. Translating these mechanistic insights, we engineered an ROS-responsive, osteoclast-targeting hydrogel for the on-demand delivery of an ARID1A-dependent canonical BRG1/BRM-associated factor complex inhibitor, which successfully alleviates TMJ-OA progression. Our findings establish the epigenetic response to ROS accumulation as a key pathogenic mechanism in TMJ-OA and highlight ARID1A as a promising therapeutic target for early disease intervention.

biomaterial(s)↗

PubMed2003

Dynamic nonlinear effect on lasing in a random medium.

We have studied both experimentally and numerically the dynamic effect of nonlinearity on lasing in disordered medium. The third-order nonlinearity not only changes the frequency and size of lasing modes, but also modifies the laser emission intensity and laser pulse width. When the nonlinear response time is longer than the lifetime of the lasing mode, the nonlinearity changes the laser output through modifying the size of the lasing mode. When the nonlinear response is faster than the buildup of the lasing mode, positive nonlinearity always extracts more laser emission from the random medium due to the enhancement of single particle scattering.

Journal Article↗

PubMed2003

Vascular endothelial growth factor inhibits outward delayed-rectifier potassium currents in acutely isolated hippocampal neurons.

In the present study, whole-cell patch-clamp recording was used to study whether vascular endothelial growth factor (VEGF) had a regulatory effect on the potassium-channel currents. The outward delayed-rectifier potassium currents (I(K)) were recorded in acutely isolated hippocampal neurons from 14-day-old rat brains. A local application of VEGF at the concentrations from 50 ng/ml to 200 ng/ml dose-dependently inhibited I(K). Administration of VEGF (100 ng/ml) to the neurons only for seconds could significantly reduce I(K) in 26 of 39 recorded cells. The currents could recover to 82.8+/-3.7% of the control level at 60 s after removing VEGF in the buffer. In the I-V curve analysis, VEGF negatively shifted the I-V curve of I(K); the inhibition was gradually enhanced as the membrane potential increased from -40 mV to 50 mV in 13 cells. Thus, the results reveal that VEGF inhibits I(K) in acute, reversible and voltage-dependent manners. Double staining combined with confocal laser scanning microscopy was used to simultaneously detect the distribution of VEGF receptors (flt-1 and flk-1) in the hippocampal section and isolated neuron. Results showed that flt-1-positive staining, but not flk-1, could be observed on the membrane of the hippocampal neuron in both preparations, suggesting the presence of neuronal membrane VEGF flt-1 receptors in the hippocampus. To investigate if the inhibition by VEGF on I(K) is related to the presence of flt-1 receptors, we further did flt-1-receptor immunostaining for the recorded neurons, which was labeled with Lucifer Yellow during the recording. Among nine recorded cells, five showing the inhibition by VEGF had detectable signals for flt-1 receptors on their membrane, whereas the other four showing no inhibition had no flt-1 receptors either. The results suggest that VEGF can acutely inhibit I(K) in the hippocampal neurons probably related to the presence of membrane flt-1 receptors in the neurons.

Action Potentials↗

PubMed2003

Multiple endocrine neoplasia type 1 (MEN1): genetic and clinical analysis in the Southern Chinese.

OBJECTIVE: Multiple endocrine neoplasia type 1 (MEN1) is characterized by a triad of neoplasia affecting the parathyroid glands, enteropancreatic endocrine tissue and the anterior pituitary gland. DESIGN: In order to define the prevalence of MEN1 germ-line mutations in Southern Chinese patients with MEN1 syndrome, we performed direct sequencing of the entire open reading frame of the MEN1 gene for 12 index patients and their first-degree relatives. RESULTS: Six patients had familial MEN1 syndrome and six had apparently sporadic disease. Nine different germ-line mutations at the MEN1 gene were identified, including three novel mutations [248-249delTT in exon 2, K559X(AAG --> TAG) in exon 10 and IVS 2nt + 2(G --> T) in intron 2]. All patients with familial MEN1 syndrome were heterozygous carriers of a germ-line mutation and MEN1-related disorders were only evident in their first-degree relatives who also carried the mutation. All patients with an enteropancreatic lesion were mutation carriers and the absence of mutation in three apparently sporadic MEN1 patients with only hyperparathyroidism and pituitary microadenoma might represent the presence of MEN1 phenocopy. CONCLUSIONS: The finding of MEN1 germ-line mutation in all patients with familial MEN1 syndrome suggests that genetic screening should be useful in our population to identify affected individuals within a kindred and allow early detection of MEN1-related tumours.

Adult↗

PubMed2002

Probing localized states with spectrally resolved speckle techniques.

We have studied the spatial extent of localized states in random media using speckle correlation techniques. Optical gain is introduced to a local region of a random medium to induce lasing of the localized states. The far-field speckle pattern of laser emission from a localized state gives its spatial field correlation function at the surface of the random medium. The envelope of the spatial field correlation function decays exponentially with the transverse coordinate on the surface. The decay length is independent of the pumping rate and the excitation area. We demonstrate that localized states exist in a disordered system that does not reach the localization threshold.

Journal Article↗

PubMed2002

Mutations in the hepatocyte nuclear factor-1alpha gene in Chinese MODY families: prevalence and functional analysis.

AIMS/HYPOTHESIS: Maturity-onset diabetes of the young is an autosomal dominant form of diabetes characterised by an early age of onset (usually <25 years). We investigated the prevalence and trans-activating activity of hepatocyte nuclear factor (HNF) -1 alpha mutations in southern Chinese families with MODY. METHODS: We screened for mutations in the HNF-1 alpha gene in 50 unrelated southern Chinese families, which fulfilled the minimum criteria for MODY. Functional properties of the mutant proteins were investigated using site-directed mutagenesis and luciferase reporter assay. RESULTS: Five of the 50 (10%) families were found to have mutations in the coding region, including a new nonsense mutation Q176X and four reported mutations (frameshift mutation P379fsdelCT, nonsense mutation R171X, missense mutations G20R and P112L). These mutations had decreased trans-activating activity on the human insulin gene promoter. We also detected a new intronic sequence variation IVS7nt-6 G-->A, which co-segregated with diabetes. The intronic variation creates a potential splice acceptor site and might alter the splicing of the HNF-1 alpha mRNA. CONCLUSION/INTERPRETATION: Mutations in the HNF-1 alpha gene seem to be an important cause of MODY in southern Chinese. The mutations could affect normal islet function by altering the expression of target genes.

Amino Acid Substitution↗

PubMed2001

Photon statistics of random lasers with resonant feedback.

We have measured the photon statistics of random lasers with resonant feedback. With an increase of the pump intensity, the photon number distribution in a single mode changes continuously from Bose-Einstein distribution at the threshold to Poisson distribution well above the threshold. The second-order correlation coefficient drops gradually from 2 to 1. By comparing the photon statistics of a random laser with resonant feedback and that of a random laser with nonresonant feedback, we illustrate very different lasing mechanisms for the two types of random lasers.

Journal Article↗

PubMed2001

Treatment of malignant digestive tract obstruction by combined intraluminal stent installation and intra-arterial drug infusion.

AIM: To study the palliative treatment of malignant obstruction of digestive tract with placement of intraluminal stent combined with intra-arterial infusion of chemotherapeutic drugs. METHODS: A total of 281 cases of digestive tract malignant obstruction were given per oral (esophagus, stomach, duodenum and jejunum), per anal (colon and rectum) and percutaneous transhepatic (biliary) installation of metallic stent. Among them, 203 cases received drug infusion by cannulation of tumor supplying artery with Seldinger's technique. RESULTS: Altogether 350 stents were installed in 281 cases, obstructive symptoms were relieved or ameliorated after installation. Occurrence of restenotic obstruction was 8-43 weeks among those with intra-arterial drug infusion, which was later than 4-26 weeks in the group with only stent installation. The average survival time of the former group was 43 (3-105) weeks, which was significantly longer than 13 (3-24) weeks of the latter group. CONCLUSION: Intraluminal placement of stent combined with intra-arterial infusion chemotherapy is one of the effective palliative therapies for malignant obstruction of the digestive tract with symptomatic as well as etiological treatment.

Adult↗

PubMed2001

Growth inhibition and apoptosis induction of Sulindac on Human gastric cancer cells.

AIM: To evaluate the effects of sulindac in inducing growth inhibition and apoptosis of human gastric cancer cells in comparison with human hepatocellular carcinoma (HCC) cells. METHODS: The human gastric cancer cell lines MKN45 and MKN28 and human hepatocellular carcinoma cell lines HepG(2) and SMMC7721 were used for the study. Anti-proliferative effect was measured by MTT assay, and apoptosis was determined by Hoechst-33258 staining, electronography and DNA fragmentation. The protein of cyclooxygenase-2 (COX-2) and Bcl-2 were detected by Western dot blotting. RESULTS: Sulindac could initiate growth inhibition and apoptosis of MKN45, MKN28, HepG(2) and SMMC7721 cells in a dose-and time-dependent manner. Growth inhibitory activity and apoptosis were more sensitive in HepG(2) cells than in SMMC7721 cells, MKN45 and MKN28 cells. After 24 hours incubation with sulindac at 2mmol x L(-1) and 4mmol x L(-1), the level of COX-2 and Bcl-2 protein were lowered in MKN45, SMMC7721 and HepG(2) cells but not in MKN28 cells. CONCLUSION: Sulindac could inhibit the growth of gastric cancer cells and HCC cells effectively in vitro by apoptosis induction, which was associated with regression of COX-2 and Bcl-2 expression. The growth inhibition and apoptosis of HCC cells were greater than that of human gastric cancer cells. The different effects of apoptosis in gastric cancer cells may be related to the differentiation of the cells.

Anti-Inflammatory Agents, Non-Steroidal↗

PubMed1999

Reversal effect of TTD on human multidrug resistant KBV200 cell line.

The reversal effect of TTD (a Chinese medicine) on human multidrug- resistant KBV200 cell line was studied and compared with verapamil (VPL). The chemosensitivity of KBV200 was detected by MTT assay in vitro and the level of MDR1 mRNA of KBV200 was investigated by RT-PCR. The cytotoxicity of TTD to KBV200 and the parent sensitive cell line KB is very low and nearly same with a concentration of 10(-6) mol x L(-1). With the concentration TTD increased VCR cytotoxicity on KBV200, 50% inhibitory concentration (IC50) decreased from 1122.5+/-72.36 mol x L(-1) to 31.76+/-5.4 nmol x L(-1) (p<0.001). It was more effective than VPL was (p<0.01). The combination of low concentration of TTD (10(-8) mol x L(-1)) and VCR (100 nmol x L(-1)) has significantly increased VCR cytotoxicity on KBV200, cell Surviving Fraction decreased from 0.91+/-0.056 to 0.74+/-0.07 (p<0.02). TTD did not inhibit the expression of MDR1 mRNA of KBV200 with the concentration of 10(-6) mol x L(-1). These data indicated that TTD could reverse VCR resistance of KBV200 and may be useful in enhancing the clinical effectiveness of VCR.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed1998

The nitric oxide/cyclic GMP system at the supraspinal site is involved in the development of acute morphine antinociceptive tolerance.

The role of the supraspinal nitric oxide (NO)/cyclic GMP system in the development of acute morphine antinociceptive tolerance was investigated by use of the mouse 55 degrees C warm-water tail-flick test. A single intracerebroventricular (i.c.v.) pretreatment of mice with morphine (3 nmol, 140 min before testing) produced an acute antinociceptive tolerance to subsequent i.c.v. doses of morphine, as demonstrated by a 120-fold rightward shift of the morphine dose-response curve. When co-administered with morphine (140 min before testing), the NO synthase inhibitors: N-nitro-L-arginine methyl ester (L-NAME), 3-bromo-7-nitroindazole, 7-nitroindazole and NG-monomethyl-L-arginine, attenuated the development of morphine tolerance. All four NO synthase inhibitors completely blocked the rightward shift of the morphine dose-response curve caused by i.c.v. morphine pretreatment (3 nmol, 140 min before testing). This effect was partially antagonized by L-arginine, but not D-arginine, in a dose-dependent manner. Also, D-NAME did not block the development of tolerance. Like the NO synthase inhibitors, LY-83,583, a guanylyl cyclase inhibitor, blocked the development of tolerance, which suggests that NO acting through the cyclic GMP pathway is involved in the development of acute antinociceptive tolerance. The effects of increased NO production on acute morphine antinociceptive tolerance were also studied. When co-administered with morphine (140 min before testing), neither L-arginine (100 nmol) nor the NO donors, sodium nitroprusside (5 nmol) and isosorbide dinitrate (10 nmol), had any effect on the magnitude of morphine antinociceptive tolerance. These results suggest that NO, acting through the cyclic GMP pathway, mediates the development of acute antinociceptive tolerance, but that NO production does not alter the magnitude of antinociceptive tolerance.

Analgesics, Opioid↗

PubMed1997

Inhibition of spinal nitric oxide synthase by N(omega)-nitro-L-arginine blocks the release of Met-enkephalin and antinociception induced by supraspinally administered beta-endorphin in the rat.

The antinociception induced by beta-endorphin given supraspinally has been demonstrated previously to be mediated by the release of Met-enkephalin acting on delta2-opioid receptors in the spinal cord. The present study was designed to determine the role of nitric oxide in the spinal cord on beta-endorphin-induced release of Met-enkephalin and antinociception. The experiments were performed in pentobarbital-anesthetized rats. The release of Met-enkephalin was performed using a spinal cord perfusion technique and the Met-enkephalin released in the spinal perfusates was measured by radioimmunoassay. Antinociception was assessed by the tail-flick test. beta-Endorphin (2 microg) given intraventricularly induced the release of Met-enkephalin from the spinal cord. The release of Met-enkephalin was dose-dependently attenuated by N(omega)-nitro-L-arginine (0.1 nM-1 microM) added into spinal perfusates and the attenuation was reversed by intrathecally applied L-arginine. The stereoisomer N(omega)-nitro-D-arginine given intrathecally, however, did not inhibit the release of Met-enkephalin induced by intraventricularly administered beta-endorphin. beta-Endorphin (4 microg) given intraventricularly produced antinociception in rats pretreated intrathecally with saline. The antinociception induced by beta-endorphin was blocked by intrathecally administered N(omega)-nitro-L-arginine (5 microg) and the blockade of antinociception was reversed by intrathecal injection of L-arginine (50 microg). N(omega)-Nitro-D-arginine (5 microg) given intrathecally did not block the intraventricularly administered beta-endorphin-induced antinociception. N(omega)-Nitro-L-arginine (10 microg) given intraventricularly did not affect intraventricularly administered beta-endorphin-induced Met-enkephalin release nor did it affect intraventricular beta-endorphin-induced antinociception, indicating that the effect of N(omega)-nitro-L-arginine is not at supraspinal sites. Intrathecal pretreatment with N(omega)-nitro-L-arginine did not affect intrathecally administered [D-Ala2]deltorphin II-induced antinociception. Our results indicate that N(omega)-nitro-L-arginine given intrathecally attenuates intraventricular beta-endorphin-administered inhibition of the tail-flick response by presynaptically inhibiting the release of Met-enkephalin.

Analgesics↗

PubMed1997

[Met]enkephalin in the spinal cord is involved in the antinociception induced by intracerebroventricularly-administered etorphine in the mouse.

We have recently reported that the antinociception induced by etorphine given i.c.v. is mediated in part by the stimulation of both mu- and epsilon-opioid receptors and the activation of both monoaminergic and opioidergic descending pain control systems. [Xu J. Y. et al. (1992) J. Pharmac. exp. Ther. 263, 246-252]. Since the opioid epsilon-receptor-mediated antinociception induced by beta-endorphin is mediated by the release of [Met]enkephalin and subsequent stimulation of delta-opioid receptors in the spinal cord, the present studies were designed to determine if beta-endorphin-like action is also involved in etorphine-induced antinociception. The tail-flick test was used to assess the antinociceptive response performed in male ICR mice. Etorphine at doses from 5 to 20 ng given i.c.v. produced a dose-dependent inhibition of the tail-flick response. The inhibition of the tail-flick response induced by etorphine given i.c.v. was antagonized by intrathecal pretreatment for 60 min with antiserum against [Met]enkephalin (10 microg), but not with antiserum against [Leu]enkephalin (10 microg) or dynorphin A (1-13) (10 microg). Desensitization of delta-opioid receptors in the spinal cord by intrathecal pretreatment with [Met]enkephalin (5 microg) for 60 min attenuated i.c.v. administered etorphine-induced tail-flick inhibition. However, intrathecal pretreatment with [Leu]enkephalin (5 microg) or dynorphin A (1-17) (0.1 microg) for 60 min did not attenuate i.c.v. administered etorphine-induced tail-flick inhibition. The results indicate that antinociception induced by etorphine given i.c.v. is mediated in part by the stimulation of the epsilon-opioid receptor at the supraspinal sites and by the release of [Met]enkephalin, which subsequently stimulates delta-opioid receptors in the spinal cord.

Analgesics, Opioid↗

PubMed1997

[Effects and mechanism of emodin and sandostatin on pancreatic ischemia in acute haemorrhagic necrotizing pancreatitis].

OBJECTIVE: To investigate pancreatic ischemia and abnormal metabolism of eicosanoids in acute haemorrhagic-necrotizing pancreatits (AHNP) and the effects of emodin or sandostatin on them. METHODS: Rats with AHNP were triggered with sodium taurocholate; the pancreatic blood flow (PBF) was detected with computerized tissue blood flowmeter, and plasma prostaglandin E2 (PGE2), 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha) and thromboxane (TXB2) were determined with radioimmunoassay. RESULTS: There was a significant decrease of PBF in the early stage of AHNP. Compared with that in the untreated group, significant improvement of PBF was demonstrated in emodin as well as in sandostatin group which showed reduced PBF following infusion of sandostatin before triggering AHNP. In untreated group plasma TXB was significantly higher, with an increase of 4.5 times, than that in sham-operated group while 6-keto-PGF1 alpha or PGE2 tended to decrease. The above mentioned abnormal synthesis of eicosanoids was blocked either in emodin or in sandostatin group in which lessened damage of acini cells was shown by pathologic scoring or transmission electron microscope. Both of the two groups shared significantly lower mortalities than the untreated group. CONCLUSION: Either emodion or sandostatin could partly reverse the decrease of PBF in the early stage of AHNP, which may be ascribed at least in part to inhibition of abnormal synthesis of eicosanoids and improvement of cytoprotection of acini cells, and combined application of the two drugs might promise positively synergetic action as well.

6-Ketoprostaglandin F1 alpha↗

PubMed1996

Tc-99m MAA total-body imaging to detect intrapulmonary right-to-left shunts and to evaluate the therapeutic effect in pulmonary arteriovenous shunts.

The appearance of radiotracer in the systemic circulation to document the visualization of the brain, kidneys, and spleen after intravenous administration of Tc-99m MAA indicates right-to-left shunts because MAA particles (20-60 microns) are supposedly trapped in the pulmonary bed (less than 15 microns). Six hypoxemia patients (1 male, 5 females; age range, 12-52 years) with intrapulmonary right-to-left shunts were evaluated by Tc-99m MAA dynamic perfusion imaging and total-body scans. Tc-99m MAA total-body imaging of the six patients with intrapulmonary right-to-left shunts (3 patients with chronic liver disease/cirrhosis of the liver and 3 patients with pulmonary arteriovenous fistulae) revealed significant radiotracer uptake in extrapulmonary organs such as the brain, kidneys, and spleen; a shunt ratio, estimated by a semiquantitative method, ranged from 17.8% to 52%. All dynamic pulmonary perfusion scans showed a normal sequence of cardiopulmonary flow without intracardiac shunts. Three patients with pulmonary arteriovenous fistulae underwent a second Tc-99m MAA total-body imaging after embolization therapy (2 patients) or lobectomy (1 patient). The result in lobectomized patients were negative for uptake in extrapulmonary organs; the two patients who underwent embolization therapy demonstrated only mild improvement. As a consequence of these findings, the authors conclude that Tc-99m MAA total-body imaging can be used for the diagnosis of intrapulmonary right-to-left shunts, as well as for the evaluation of postshunt therapy.

Adult↗

PubMed1996

[Studies on the bioactive constituents of Aurantii Fructus Immaturus].

An ethanol extract of "Kijitsu" (Aurantii Fructus Immaturus, Citrus aurantium L.) collected in China was assessed for the antitumor activity using murine leukemia P388 in vivo, and the extract was found to be active by the antitumor bioassay in vivo and in vitro. The extract was separated into a petroleum ether-soluble fraction and an ethyl acetate-soluble fraction. Fractionation was carried out using an index of cell-growth inhibitory activity against mouse leukemia L1210 cells to isolate antitumor active substances or compounds. The active compounds were purified employing silica gel column chromatography and HPLC. The antitumor effect of the isolated active compounds was studied. Five compounds, auraptene, marmin, tangeretin, nobiretin and 5-[(6',7'-dihydroxy-3',7'-dimethyl-2-octenyl)oxy]psoralen were isolated from Citrus aurantium L. Though they are all known compounds, 5-(6',7'-dihydroxy-3',7-dimethyl-2-octenyl)oxy-psoralen from this plants was first isolated. These compounds showed a cell-growth inhibitory effect against L1210 and K562 in vitro.

Animals↗

PubMed1996

[Synergism of sobuzoxane in combination with doxorubicin against leukemia P388 in mice].

AIM: To study the antitumor activity of sobuzoxane (Sob) in combination with doxorubicin (Dox) and the effect of Sob on Dox-induced cardiotoxicity. METHODS: DBA/2 mice bearing transplanted leukemia P388 were given i.v. Dox 2 mg.kg-1.d-1 for 3 d, 4 mg.kg-1.d-1 for 1 d combined with ig Sob 20, 40 mg.kg-1.d-1 for 7 d. The increase in life span (ILS) of each group was recorded in 30 d. The myocardium of moribund mice was examined by transmission electron microscopy. RESULTS: The ILS of combination therapeutic groups of Sob with Dox was 48.7%, 57.3%, 59.0%, and 62.4% respectively, which were 30%-90% higher than the sum of ILS of two groups treated with Dox and Sob separately (P < 0.01). The ultrastructural injury of cardiomyocytes of P388-bearing mice caused by combination chemotherapy with Dox plus Sob was markedly attenuated compared with Dox alone. CONCLUSION: Sob with Dox exhibited an antitumor synergistic effect on leukemia P388, and the cardiotoxicity of Dox was reduced by Sob.

Animals↗

PubMed1996

N-cyclobutylmethyl analog of normorphinone, N-CBM-TAMO: a short-term opioid agonist and long-term Mu-selective irreversible opioid antagonist.

The antinociceptive and opioid binding properties of the N-cyclobutylmethyl analog of normorphinone, 14 alpha, 14' beta-[dithiobis[(2-oxo-2, 1-ethanediyl)imino]]bis[7,8-dihydro-N-(cyclobutylmethyl)-normor phinone] (N-CBM-TAMO) were investigated. This compound is a dimer, containing a disulfide capable of binding to thiol groups on the opioid receptor. Competition radioligand binding assays with bovine striatal membranes demonstrated that N-CBM-TAMO displayed a higher affinity for mu opioid receptors than for kappa and delta receptors. Incubation of membranes with N-CBM-TAMO resulted in wash-resistant inhibition of the binding of the mu-selective peptide [3H][D-Ala2,(Me)Phe4, Gly(ol)5]-enkephalin, the kappa-selective opioid [3H]U69,593 ((trans)-3, 4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl)-cyclohexyl]benzenacetamide+ ++ methanesulfonate hydrate)) and, to a lesser extent, the delta-selective peptide [D-Pen2, p-Cl-phenylalanine4, D-Pen5]enkephalin. Scatchard analysis of saturation binding data showed that N-CBM-TAMO decreased the Bmax value without affecting the Kd value for [3H][D-Ala2,(Me)Phe4, Gly(ol)5]enkephalin binding, whereas, N-CBM-TAMO increased the Kd value without altering the Bmax value for [3H]U69,593, which suggests that N-CBM-TAMO interacted covalently with the mu but not the kappa receptor. In the mouse 55 degrees C warm-water tail-flick test, N-CBM-TAMO given supraspinally acted as an agonist with low efficacy because only submaximal antinociception was observed at doses up to 100 nmol. The antinociception induced by N-CBM-TAMO in the tail-flick test was partially blocked by both the mu-selective antagonist beta-funaltrexamine and the kappa-selective antagonist nor-binaltorphimine. In the mouse acetic acid writhing test, N-CBM-TAMO acted as a full agonist with a D50 value of 0.08 (0.04-0.14) nmol, and the antinociception was blocked by coadministration of the kappa-selective antagonist nor-binaltorphimine. Pretreatment of mice with an i.c.v. dose of N-CBM-TAMO of 10 nmol, a dose that exhibited modest short-term antinociception in the tail-flick test, produced a time- and dose-dependent long-term antagonism of morphine-induced antinociception in an irreversible manner in this assay. Pretreatment of mice with i.c.v. N-CBM-TAMO at doses of 3 nmol and higher, which produced supermaximal short-term antinociception in the writhing test, produced a time- and dose-dependent long-term antagonism of U50,488 (trans)-3, 4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl)-cyclohexyl]benzeneacetamide methanesulfonate hydrate)-induced antinociception in a reversible manner, probably because of the development of tolerance. These in vivo data, together with the in vitro binding data, demonstrate that N-CBM-TAMO is a potent kappa agonist and at higher doses produces antinociception mediated by mu receptors. N-CBM-TAMO also produces long-term noncompetitive antagonism of antinociception mediated by mu opioid receptors.

Analgesics, Opioid↗