Methionine Enkephalin Research Articles

Pharmaceutical Excipients Enhance Iron-Dependent Photo-Degradation in Pharmaceutical Buffers by near UV and Visible Light: Tyrosine Modification by Reactions of the Antioxidant Methionine in Citrate Buffer.

To evaluate the effect of excipients, including sugars and amino acids, on photo-degradation reactions in pharmaceutical buffers induced by near UV and visible light. Solutions of citrate or acetate buffers, containing 1 or 50μM Fe3+, the model peptides methionine enkephalin (MEn), leucine enkephalin (LEn) or proctolin peptide (ProP), in the presence of commonly used amino acids or sugars, were photo-irradiated with near UV or visible light. The oxidation products were analyzed by reverse-phase HPLC and HPLC-MS/MS. The sugars mannitol, sucrose and trehalose, and the amino acids Arg, Lys, and His significantly promote the oxidation of peptide Met to peptide Met sulfoxide. These excipients do not increase the yields of hydrogen peroxide, suggesting that other oxidants such as peroxyl radicals are responsible for the oxidation of peptide Met. The addition of free Met reduces the oxidation of peptide Met, but, in citrate buffer, causes the addition of Met oxidation products to Tyr residues of the target peptides. Commonly used excipients enhance the light-induced oxidation of amino acids in model peptides.

Effects of Pulsed Radiofrequency with Different Temperature on Model Rats of Chronic Constriction Injury

The treatment for neuropathic pain is still a big challenge. Pulsed radiofrequency technique has been widely used to relieve neuropathic pain in recent years. The purpose of this study is to optimize the temperature for pulsed radiofrequency therapy. Animal, experimental study. Seventy-five male SD rats were randomly divided into five groups: Sham operation group (Sham group), chronic constriction injury group (CCI group), PRF 42°C group (P42 group), PRF 50°C group (P50 group), and PRF 60°C group (P60 group). The hindpaw withdrawal threshold (HWT), paw thermal withdrawal latency (PTWL), sciatic nerve structure, and the concentration of spinal methionine enkephalin(M-ENK) were detected to identify which temperature is the best for PRF treatment. PRF at 42°C, 50°C and 60°C significantly alleviated the pain in CCI rats. The therapeutic effects of 50°C and 60°C were similar, and both were better than 42°C. In addition, PRF using 42°C, 50°C, and 60°C mediated nerve injury to sciatic nerve were grade 1, 1, and 2, respectively. The concentration of M-ENK in spinal cord increased accompanying with the increasing of the temperature of PRF. PRF using 50°C could induce less damage while achieving better improvement of mechanical and thermal pain threshold than 42°C and 60°C in CCI rats, which may be achieved by promoting the expression of M-ENK in spinal cord.

Alkali metal ion-induced conformation changes of methionine- and leucine enkephalin investigated by gas-phase hydrogen/deuterium exchange combined with theoretical calculations

We have investigated the effects of alkali-metal ions (G+ = K+, Rb+, or Cs+) on the conformation of leucine enkephalin (LE) and methionine enkephalin (ME) using hydrogen/deuterium exchange mass spectrometry and theoretical calculations at the molecular level. The MS result revealed a 1:1 stoichiometric ratio of the G+ coordination with LE/ME. We used [LE/ME+H]+ as the native conformation for comparison purposes, we observed different HDX behaviors following the reactivity order [LE/ME+H]+>[LE/ME+K]+>[LE/ME+Rb]+>[LE/ME+Cs]+ and [LE+G]+>[ME+G]+. Eight hydrogen atoms were HD-exchanged for [LE/ME+H]+, whereas only 4, 3, and 2 hydrogen atoms underwent HDX for [LE/ME+K]+, [LE/ME+Rb]+, and [LE/ME+Cs]+, respectively. Additionally, we obtained different dissociation processes and different fragmentations for the various complexes, and their varied HDX behaviors suggest that the alkali-metal ions K+, Rb+, and Cs+ produce effects on the LE/ME conformations that differ from those of H+. Further, we investigated the favorable conformations of the complexes by theoretical calculations, and found that K+, Rb+, and Cs+ are coordinated with three carbonyl oxygens and one deprotonated carboxylic acid of LE/ME, whereas the terminal amino group is the most stable site for protonation. We calculated the proton affinity (PA) for each active hydrogen atom on the optimized complexes, which reasonably explained the HDX results that a high PA value corresponds to low HDX reactivity. Overall, the calculations showed that alkali-metal ions can induce ME into a more rigid conformation than LE. Even the native conformation of ME is more exposed than LE, and the conformation of LE/ME is stabilized and rigid owing to the increase in the size of the complexed alkali metals.

Probing Structural Information of Gas-Phase Peptides by Near-Edge X-ray Absorption Mass Spectrometry.

Near-edge X-ray absorption mass spectrometry (NEXAMS) is an action-spectroscopy technique of growing interest for investigations into the spatial and electronic structure of biomolecules. It has been used successfully to give insights into different aspects of the photodissociation of peptides and to probe the conformation of proteins. It is a current question whether the fragmentation pathways are sensitive toward effects of conformational isomerism, tautomerism, and intramolecular interactions in gas-phase peptides. To address this issue, we studied the cationic fragments of cryogenically cooled gas-phase leucine enkephalin ([LeuEnk+H]+) and methionine enkephalin ([MetEnk+H]+) produced upon soft X-ray photon absorption at the carbon, nitrogen, and oxygen K-edges. The interpretation of the experimental ion yield spectra was supported by density-functional theory and restricted-open-shell configuration interaction with singles (DFT/ROCIS) calculations. The analysis revealed several effects that could not be rationalized based on the peptide's amino acid sequences alone. Clear differences between the partial ion yields measured for both peptides upon C 1s → π*(C═C) excitations in the aromatic amino acid side chains give evidence for a sulfur-aromatic interaction between the methionine and phenylalanine side chain of [MetEnk+H]+. Furthermore, a peak associated with N 1s → π*(C═N) transitions, linked to a tautomeric keto-to-enol conversion of peptide bonds, was only present in the photon energy resolved ion yield spectra of [MetEnk+H]+.

Study on the mechanism of the Pu-erh tea (Camellia sinensis var. assamica) extract inhibiting contraction of isolated mouse duodenum

The aims of this study were to investigate the effects of three different concentrations of the Pu-erh tea extract (PTE) on the contractile activity of the isolated mouse duodenum and explore their mechanism. The contraction amplitude and frequency of the isolated mouse duodenum were inhibited by all three concentrations of PTE. The high-concentration PTE significantly (P < 0.01) inhibited the promotion effects of acetylcholine chloride or BayK8644 on the amplitude and frequency of intestinal contraction, which were comparable to those of atropine sulphate and verapamil hydrochloride, respectively. The results of UV-Vis and ELISA showed that the content of methionine-enkephalin (Met-ENK) in the PTE-treated groups was decreased to varying degrees; contrarily, the activities of tyrosine hydroxylase (TH), total nitric oxide synthase, and the content of nitric oxide were increased to different degrees. The results suggest that PTE can inhibit the contraction of the isolated mouse duodenum, and the mechanism of action is that PTE can not only inhibit the signal transduction pathways of AC-cAMP-PKA and PLC-IP3-Ca2+, but also the Ca2+ signal systems mediated by G protein-coupled M receptors through the myenteric plexus. By reducing the release of Met-ENK from the motor neurons of the myenteric plexus, the GTP-cAMP-PKK signalling pathway and the Ca2+ signalling system mediated by G protein-coupled delta receptors were inhibited. By increasing the TH activity of the motor neurons in the myenteric plexus, the norepinephrine content was increased, thereby the AC-cAMP-PKA signal transduction pathway mediated by G protein-coupled β receptors was activated. This study increases knowledge regarding the medicinal value of the Pu-erh tea.

Can eosinophils in adipose tissue add fuel to the fire?

In inguinal adipose tissue, beige adipocytes are interspersed among white adipocytes and in close communication with dynamic populations of immune cells. Recent data have demonstrated that anti-inflammatory macrophages (M2) increase thermogenic activity of beige adipocytes, although the mechanism is currently under debate. ILC2, cells via secretion, of methionine enkephalin peptide were found to be able to increase beige thermogenesis. Knights et al. have recently generated a whole-body knock-out of KLF3 (KLF3-/-) to explore its contribution to thermogenesis and weight gain in a diet-induced obesity animal model.

Effects of botulinum toxin A on endometriosis‑associated pain and its related mechanism.

Endometriosis (EMS) is a common disease in women aged 25–45 years, and pain is the main clinical symptom. The primary clinical treatment is surgical excision and drug therapy targeting the ectopic lesions, but these have not been very effective. Botulinum neurotoxin serotype A (BTX-A) has been reported to be useful in the treatment of pain in a variety of diseases. Based on this, the aim of the present study was to explore the therapeutic effect and mechanism of BTX-A on EMS. A model of nerve injury induced by oxygen glucose deprivation (OGD) was constructed in PC12 cells and EMS mice. Model cells and mice were treated with different concentrations of BTX-A to observe the changes in pain behavior, to detect cell viability and the secretion of norepinephrine (NE) and methionine enkephalin (M-EK) in cells and the spinal cord, and to evaluate the expression of apoptosis-related molecules in spinal cord nerves. The results revealed that BTX-A significantly reduced the amount of writhing in model mice, enhanced the activity of PC12 OGD cells, increased the secretion of NE and M-EK in model cells and the spinal cord of mice, and decreased the apoptosis of neural cells in the spinal cord of the model mice. Therefore, it was hypothesized that BTX-A may alleviate the pain induced by EMS by increasing the secretion of analgesic substances and promoting the repair of nerve injury. The present study provided a theoretical basis for the treatment of pain induced by EMS.

Glutathione and Glutathione-Like Sequences of Opioid and Aminergic Receptors Bind Ascorbic Acid, Adrenergic and Opioid Drugs Mediating Antioxidant Function: Relevance for Anesthesia and Abuse.

Opioids and their antagonists alter vitamin C metabolism. Morphine binds to glutathione (l-γ-glutamyl-l-cysteinyl-glycine), an intracellular ascorbic acid recycling molecule with a wide range of additional activities. The morphine metabolite morphinone reacts with glutathione to form a covalent adduct that is then excreted in urine. Morphine also binds to adrenergic and histaminergic receptors in their extracellular loop regions, enhancing aminergic agonist activity. The first and second extracellular loops of adrenergic and histaminergic receptors are, like glutathione, characterized by the presence of cysteines and/or methionines, and recycle ascorbic acid with similar efficiency. Conversely, adrenergic drugs bind to extracellular loops of opioid receptors, enhancing their activity. These observations suggest functional interactions among opioids and amines, their receptors, and glutathione. We therefore explored the relative binding affinities of ascorbic acid, dehydroascorbic acid, opioid and adrenergic compounds, as well as various control compounds, to glutathione and glutathione-like peptides derived from the extracellular loop regions of the human beta 2-adrenergic, dopamine D1, histamine H1, and mu opioid receptors, as well as controls. Some cysteine-containing peptides derived from these receptors do bind ascorbic acid and/or dehydroascorbic acid and the same peptides generally bind opioid compounds. Glutathione binds not only morphine but also naloxone, methadone, and methionine enkephalin. Some adrenergic drugs also bind to glutathione and glutathione-like receptor regions. These sets of interactions provide a novel basis for understanding some ways that adrenergic, opioid and antioxidant systems interact during anesthesia and drug abuse and may have utility for understanding drug interactions.

Proenkephalin+ regulatory T cells expanded by ultraviolet B exposure maintain skin homeostasis with a healing function

Regulatory T (Treg) cells, expressing CD25 (interleukin-2 receptor α chain) and Foxp3 transcription factor, maintain immunological self-tolerance and suppress various immune responses. Here we report a feature of skin Treg cells expanded by ultraviolet B (UVB) exposure. We found that skin Treg cells possessing a healing function are expanded by UVB exposure with the expression of an endogenous opioid precursor, proenkephalin (PENK). Upon UVB exposure, skin Treg cells were expanded with a unique TCR repertoire. Also, they highly expressed a distinctive set of genes enriched in "wound healing involved in inflammatory responses" and the "neuropeptide signaling pathway," as indicated by the high expression of Penk. We found that not only was PENK expression at the protein level detected in the UVB-expanded skin Treg (UVB-skin Treg) cells, but that a PENK-derived neuropeptide, methionine enkephalin (Met-ENK), from Treg cells promoted the outgrowth of epidermal keratinocytes in an ex vivo skin explant assay. Notably, UVB-skin Treg cells also promoted wound healing in an in vivo wound closure assay. In addition, UVB-skin Treg cells produced amphiregulin (AREG), which plays a key role in Treg-mediated tissue repair. Identification of a unique function of PENK+ UVB-skin Treg cells provides a mechanism for maintaining skin homeostasis.

A close neuroanatomical relationship between the enkephalinergic (methionine-enkephalin) and tachykininergic (substance P) systems in the alpaca diencephalon.

In the alpaca diencephalon, the distribution of immunoreactive cell bodies and fibers containing methionine-enkephalin (MET) or substance P (SP) has been studied. The immunohistochemical study was performed by standard method on the diencephalon of four male alpacas that lived at sea level. Nerve fibers containing MET or SP were widely distributed in the thalamus and hypothalamus. METand SP-immunoreactive fibers showed a similar distribution in the whole diencephalon. Immunoreactive cell bodies containing MET or SP were only observed in the hypothalamus. The distribution of MET-immunoreactive cell bodies was more widespread than that observed for cell bodies containing SP. A close neuroanatomical relationship between the tachykininergic (SP) and enkephalinergic (MET) systems was observed in the whole diencephalon suggestive of the existence of multiple physiological interactions between both systems.

Methionine Enkephalin Suppresses Osteocyte Apoptosis Induced by Compressive Force through Regulation of Nuclear Translocation of NFATc1.

ABSTRACTMechanical stress stimulates bone remodeling, which occurs through bone formation and resorption, resulting in bone adaptation in response to the mechanical stress. Osteocytes perceive mechanical stress loaded to bones and promote bone remodeling through various cellular processes. Osteocyte apoptosis is considered a cellular process to induce bone resorption during mechanical stress‐induced bone remodeling, but the underlying molecular mechanisms are not fully understood. Recent studies have demonstrated that neuropeptides play crucial roles in bone metabolism. The neuropeptide, methionine enkephalin (MENK) regulates apoptosis positively and negatively depending on cell type, but the role of MENK in osteocyte apoptosis, followed by bone resorption, in response to mechanical stress is still unknown. Here, we examined the roles and mechanisms of MENK in osteocyte apoptosis induced by compressive force. We loaded compressive force to mouse parietal bones, resulting in a reduction of MENK expression in osteocytes. A neutralizing connective tissue growth factor (CTGF) antibody inhibited the compressive force‐induced reduction of MENK. An increase in osteocyte apoptosis in the compressive force‐loaded parietal bones was inhibited by MENK administration. Nuclear translocation of NFATc1 in osteocytes in the parietal bones was enhanced by compressive force. INCA‐6, which inhibits NFAT translocation into nuclei, suppressed the increase in osteocyte apoptosis in the compressive force‐loaded parietal bones. NFATc1‐overexpressing MLO‐Y4 cells showed increased expression of apoptosis‐related genes. MENK administration reduced the nuclear translocation of NFATc1 in osteocytes in the compressive force‐loaded parietal bones. Moreover, MENK suppressed Ca2+ influx and calcineurin and calmodulin expression, which are known to induce the nuclear translocation of NFAT in MLO‐Y4 cells. In summary, this study shows that osteocytes expressed MENK, whereas the MENK expression was suppressed by compressive force via CTGF signaling. MENK downregulated nuclear translocation of NFATc1 probably by suppressing Ca2+ signaling in osteocytes and consequently inhibiting compressive force‐induced osteocyte apoptosis, followed by bone resorption. © 2020 The Authors. JBMR Plus published by Wiley Periodicals, Inc. on behalf of American Society for Bone and Mineral Research.

Roles of Neurotransmitters in Decapod Reproduction

In decapods neurotransmitters govern a variety of physiological events including growth, moulting, reproduction, locomotor activity, behavior and sexual differentiation. The most important reproductive neurotransmitters in decapods are histamine (HA), dopamine (DA), 5-hydrtryptamine (5HT, serotonin), leucine enkephalin (Leu-ENK), octopamine (OA), epinephrine (EPI), methionine enkephalin (Met-ENK), and Red pigment-concentrating hormone (RPCH). In this study, the discovery and identification of reproductive neurotransmitters in decapods are reviewed. The role of neurotransmitters (i.e. 5-HT, DA, OA, RPCH, Leu-ENK and Met-ENK) on the reproduction and sexual maturation of decapods are remarkable. It can be concluded that understanding the roles and effects of neurotransmitters are useful for further basic studies in reproductive biology of decapodsas well as commercial profitability in terms of development of new techniques for assisted reproduction in aquaculture.

Methionine-enkephalin-treatment suppresses the pituitary-ovary axis in the cichlid fish Oreochromis mossambicus

The involvement of endogenous opioid peptides in regulation of reproductive activity is implicated in a majority of vertebrates. Methionine-enkephalin (M-ENK) is an endogenous opioid peptide neurotransmitter produced in the central nervous system. Although a role for M-ENK in antinociception (pain suppression) and mediation of stress has been suggested, its influence on fish reproduction is not clearly understood. The aim of the present investigation was to determine the role of M-ENK on the reproductive-axis of the female tilapia Oreochromis mossambicus. Treatment of stripped O. mossambicus with 300 μg M-ENK during the ovarian cycle resulted in suppression of luteinizing hormone (LH) secretory activity. This was evidenced by a significant reduction in the area and the intensity of LH immunoreactivity in the proximal pars distalis of the pituitary gland as compared to controls during the prespawning phase. At the ovarian level, the treatment of M-ENK caused a significant increase in the mean number of follicles with chromatin nucleolar (stage I), perinucleolar (stage II) and cortical alveolar stages (Stage III). Although the mean number of early vitellogenic follicles (stage IV) did not significantly differ from that of initial controls and controls, the mean number of late vitellogenic follicles (stage V, preovulatory) and the serum levels of estradiol were significantly lower in M-ENK treated fish compared to those of controls. However, there was a significant increase in the serum levels of cortisol in M-ENK treated fish compared to controls. Together, these results suggest that treatment of M-ENK inhibits the recruitment of follicles at late vitellogenic stage through the suppression of steroidogenesis in the ovary and LH secretory activity at the level of the pituitary gland in fish. Besides, the ability of M-ENK to stimulate serum cortisol levels indicates a role for this peptide in regulation of the hypothalamo-pituitary-interrenal axis in fish.

Site-Selective Dissociation upon Sulfur L-Edge X-ray Absorption in a Gas-Phase Protonated Peptide.

Site-selective dissociation induced by core photoexcitation of biomolecules is of key importance for the understanding of radiation damage processes and dynamics and for its promising use as "chemical scissors" in various applications. However, identifying products of site-selective dissociation in large molecules is challenging at the carbon, nitrogen, and oxygen edges because of the high recurrence of these atoms and related chemical groups. In this paper, we present the observation of site-selective dissociation at the sulfur L-edge in the gas-phase peptide methionine enkephalin, which contains only a single sulfur atom. Near-edge X-ray absorption mass spectrometry has revealed that the resonant S 2p → σ*C-S excitation of the sulfur contained in the methionine side chain leads to site-selective dissociation, which is not the case after core ionization above the sulfur L-edge. The prospects of such results for the study of charge dynamics in biomolecular systems are discussed.

Methionine enkephalin inhibits influenza A virus infection through upregulating antiviral state in RAW264.7 cells

MENK, as an immune adjuvant, has potential immune-regulatory activity on innate and adaptive immune cells. The aim of this work was to investigate the antiviral effect of MENK on influenza virus-infected murine macrophage cells (RAW264.7) and its underlying mechanisms. The results showed that MENK markedly inhibited influenza A virus (H1N1) replication in pre- and post-MENK treatment, especially in pre-MENK treatment. The mechanisms exploration revealed that MENK (10 mg/mL) significantly inhibited the nucleoprotein (NP) of influenza virus and up-regulated levels of IL-6, TNF-α and IFN-β compared with those in H1N1 control group. Further experiments confirmed that antiviral effects of MENK was associated with promotion of opioid receptor (MOR) as well as activation of NF-κB p65 inducing cellular antiviral status. The data suggest that MENK should be potential candidate for prophylactic or therapeutic treatment against H1N1 influenza virus.

Characterization of Union Sites for Opioids in Human Lymphocyte Membranes

The opioid peptides Methionine enkephalin and β-Endorphin are stimulators of Natural Killer cytolytic activity. To know the biochemical mechanisms that explain this action, the binding of Methionine enkephalin and Naloxone in presence or ausence of Na+ ion and GTP to human Lymphocyte membranes was characterized.

Methionine enkephalin (MENK) regulates the immune pathogenesis of type 2 diabetes mellitus via the IL-33/ST2 pathway

The incidence and mortality of type 2 diabetes mellitus (T2DM) rank among the top ten worldwide. Emerging studies indicate pathological roles for the immune system in inflammation, insulin resistance and islet β-cell damage in subjects with T2DM. Methionine enkephalin (MENK) is present in endocrine cells of the pancreas and has been suggested to be an important mediator between the immune and neuroendocrine systems. Therefore, it may play a role in modulating insulin secretion from islet cells. Since little is known about the effect of MENK on T2DM, therefore it was the aim of this study to characterize the role and possible mechanism of action of MENK on plasma glucose and serum insulin levels in T2DM rats and INS-1 cells in vivo and in vitro. MENK significantly decreased the plasma glucose level and increased the serum insulin concentration in T2DM rats. It also increased the serum levels of the cytokines IL-5 and IL-10, while decreased TNF-α and IL-2 levels. We further confirmed that MENK regulated glucose metabolism by upregulating opioid receptor expression and modulating the IL-33/ST2 and MyD88-TRAF6-NF-κB p65 signaling pathways. Based on these results, an intraperitoneal injection of MENK represents a potentially new approach for T2DM.

Characterization of a Multiple-Scan-Rate Voltammetric Waveform for Real-Time Detection of Met-Enkephalin.

Opioid peptides are critically involved in a variety of physiological functions necessary for adaptation and survival, and as such, understanding the precise actions of endogenous opioid peptides will aid in identification of potential therapeutic strategies to treat a variety of disorders. However, few analytical tools are currently available that offer both the sensitivity and spatial resolution required to monitor peptidergic concentration fluctuations in situ on a time scale commensurate with that of neuronal communication. Our group has developed a multiple-scan-rate waveform to enable real-time voltammetric detection of tyrosine containing neuropeptides. Herein, we have evaluated the waveform parameters to increase sensitivity to methionine-enkephalin (M-ENK), an endogenous opioid neuropeptide implicated in pain, stress, and reward circuits. M-ENK dynamics were monitored in adrenal gland tissue, as well as in the dorsal striatum of anesthetized and freely behaving animals. The data reveal cofluctuations of catecholamine and M-ENK in both locations and provide measurements of M-ENK dynamics in the brain with subsecond temporal resolution. Importantly, this work also demonstrates how voltammetric waveforms can be customized to enhance detection of specific target analytes, broadly speaking.

Methionine enkephalin (MENK) inhibits human gastric cancer through regulating tumor associated macrophages (TAMs) and PI3K/AKT/mTOR signaling pathway inside cancer cells

This study was to explore the effect and mechanisms of anti- human gastric cancer by MENK in vitro and in vivo. The results showed in MENK-treated xenograft tissue, the percentage of M2-type macrophages decreased while M1-type macrophages increased. MENK increased the expression of M1-related cytokine TNF-α and attenuated the expression of M2-related cytokine IL-10 expression.MENK upregulated the expression of opioid receptor (OGFr), while it inhibited HGC27 and SGC7901 cells through blocking PI3K/AKT/mTOR signal pathway in vitro and in vivo. These effects of MENK could be cancelled when OGFr was knockdown. This indicates that binding to OGFr by MENK appears to be essential for the anti- GC cells. Therefore, it is concluded that MENK might skew macrophage toward M2 phenotype from M1 phenotype within tumor and induce cells apoptosis though blocking OGFr/PI3K/AKT/mTOR signaling pathway.

Rules to activate CD8+T cells through regulating subunits of opioid receptors by methionine enkephalin (MENK)

The goal of this work was to investigate how MENK could regulate the functions of CD8+T cells and to explore the relationship between this regulation and opioid receptor expression. Our results showed that the opioid receptors presented on the cell menbrane of CD8+T cells were MOR and DOR. MENK promoted the expression of opioid receptors as well as the elevation of the surface molecules such as CD28, PD-1, CTLA-4 and FasL and intracellular granzyme B. Selectively blocking the MOR by CTAP or DOR by NTI could result in inhibition of the corresponding CD8+T cells proliferation and the expressions of surface molecules. In addition, non-selectively blocking both MOR and DOR by NTX could further impair the functions and proliferation of CD8+T cells. Our currently data indicated that MENK could play a vital role in immune functions via precise regulation to subunits of opioid receptors.