1.The function of C-terminal tail of opioid receptors and the proteins interaction with C-terminal tail of opioid receptors
Peilan ZHOU ; Lingdi YAN ; Zehui GONG
Chinese Pharmacological Bulletin 2003;0(08):-
The C-terminal tail of opioid receptor is important for opioid receptor phosphorylation,desensitization and internalization after opioid agonists treatment.The C-terminal point mutant and truncated mutant of amino acid residues of ?opioid receptors such as T394A,T383A,T357A,S355A affect the function of the opioid receptor obviously.The identical results could be seen in the C-terminal point mutant S363A of ?opioid receptor and S369A of ?opioid receptor.It is well known that opioid receptor requires interaction with other proteins for function,regulation and trafficking.Although proteins such as PPL,FilaminA,PLD2,PKCI,GASP and EBP50/NHERF have been identified to interact with C-terminals of opioid receptors,the role of interaction of these proteins with opioid receptor in the dependence,tolerance and addiction of opioids is dubious.Therefore,looking for the proteins specifically interacting with the C-terminals of opioid receptor may be important to understand the mechanism of opioids addiction.
2.Analysis of 28 Fatal Cases Caused by Drug Adverse Reaction
Peilan DENG ; Honglan ZHOU ; Jun FENG
China Pharmacy 2001;12(6):367-
OBJECTIVE: To study the disease distribution and lethal factors of ADRs in 28 fatal cases,so as to improve the prophylactic measures for diseases and rationality and therapeutic effect of drug-use.METHODS: According to ADRs evaluation standard,the cases who died in this hospital in recent 2 years were analysed with computer internet.RESULTS: 28 cases directly died of ADRs.Insulin and glibenclamidium were the most common lethal agents,accounting for 32.14% ,and mannitol and azathiopurine were the next ones.Senile patient accounted for 63.28% of all patients died of ADRs.CONCLUSION: (1)Rational drug-use has clinical significance in reducing mortality of geratic diseases.(2)It will be an important way in reducing the fatality of ADRs that we inquire about patients' illness carefully and handle the indications and dosages of drugs seriously.
3.Establishment of a recombinant cell line that stably expresses rat purinergic receptor P2X4 and verification of its function
Linlin LI ; Peilan ZHOU ; Ruibin SU
Chinese Journal of Pharmacology and Toxicology 2014;(3):426-431
OBJECTIVE Toestablisharecombinantcelllinethatcanstablyexpressratpurinergic P2receptorP2X4(rP2X4R).METHODS ToconstructgreenfluorescentproteinandrP2X4recombinant plasmid (pEGFP-N1-rP2X4),lipofectamine was used to transfect pEGFP-N1-rP2X4 into human embry-onic kidney (HEK293)cells that were screened with G41 8 (1 g·L-1 ).The quantitative expression of rP2X4 receptor was verified by qRT-PCR and Western blotting analysis.Whole-cell patch clamp record-ing was used to investigate the function of the stably expressed rP2X4 receptor. RESULTS The sequence of plasmid pEGFP-N1-rP2X4 was verified by PubMed Blastn comparison.qRT-PCR and Western blotting analysis demonstrated that the expression of rP2X4 receptor in HEK293-pEGFP-N1-rP2X4 cell lines remained stable after 25 generations (P1 ,P3,P5,P10,P15,P20 and P25).Whole-cell patch clamp recording experiments showed that the rP2X4 receptor agonist,purine-5′-triphosphate (ATP,3.0 μmol·L-1 ),could activate rP2X4 receptors in HEK293-pEGFP-N1-rP2X4 cell lines.Specific activating current could be blocked by non-selective rP2X4 receptor antagonist TNP-ATP (30.0μmol·L-1).CONCLUSION rP2X4receptorisstablyexpressedinHEK293-pEGFP-N1-rP2X4cell line and maintains stable expression and function within 25 continuous generations.The establish ment of HEK293-pEGFP-N1-rP2X4 cell line can contribute to further investigations of the roles of rP2X4 receptors in neuropathic pain.
4.Mechanism of opioid receptor ubiquitination and its effect on receptor function
Jiebing JLANG ; Peilan ZHOU ; Zhibing ZHENG ; Ruibin SU
Chinese Journal of Pharmacology and Toxicology 2015;(2):302-309
Opioid receptors, as an important member of G protein coupled receptors (GPCR), are the binding targets of endogenous opioid peptides and exogenous opiates. The activation of opioid receptors influences the nervous system, immune physiology and endocrine system. However, prolonged activation of opioid receptors is likely to produce opioid tolerance, leading to opioid addiction. Receptor endocytosis and sorting into the recycling pathway contribute to recovery of cellular opioid responsiveness. Recent studies have revealed that GPCR can be modulated by ubiquitination which plays a unique roles in governing GPCR trafficking. Moreover, ubiquitination of the opioid receptors (μ, κand δ) is increased after stimulation of most opioid agonists. Mutation of the ubiquitin sites affects the internalization and degradation of opioid receptors, which contributes to changes in signal pathways and regulation of opioid receptors. ln this paper, ubiquitination of opioid receptors and the fundamental role of ubiquitination in trafficking of opioid receptors are reviewed.
5.Study on Jangzhuo Decotion for Treating Plasma Concenration of Oxidized Low Density Lipoprotein D Dimer and Serum Lipid of Ischemic Stroke and Its Relation to Preventing Repeated Stroke
Ronggen ZHOU ; Peilan YANG ; Lei ZHAO ; Xiaofen SHI ;
Chinese Traditional Patent Medicine 1992;0(01):-
Objective: Clinical study on Jangzhuo Decoction for treating plasma concentrations of oxidized low density lipoprotein, d dimer of ischemic stroke and its relation to preventing repeated stroke.Methods: 72 cases with ischemic stroke were divided randomly and blindly into two groups. 36 patients with ischemic stroke in therapeutic group were treated with Jangzhuo Decoction M ASA, and 36 patients in control group were treated with M ASA alone. The follow up survey for six months was conducted when physical signs and symptoms of the paints with ischemic stroke begain to be stable. Results: The incidence of the repeated stroke in therapeutic group were one case(2.7%) and in control group were three cases(8.3%) respectively. Ox LDL levels of therapeutic group was 1021?102?g/dl( ?s ), decreased to 604?77?g/dl( P 0.05). There was significant difference between the two groups. D dimer levels of two grouups were all increased significantly( P
6.Role of histamine H3 receptor in regulation of hypoxia
Jingjing LIAN ; Lingdi YAN ; Peilan ZHOU ; Ruibin SU
Chinese Journal of Pharmacology and Toxicology 2016;30(4):369-374
Hypoxia refers to the reduction in tissue oxygen supply or utilization. It occurs in various pathological symptoms like embolism,anthracemia,and chronic obstructive sleep. At high altitude, lower partial pressure of oxygen compromises the supply of adequate oxygen to the tissues and leads to many clinical syndromes,such as acute mountain sickness,high-altitude cerebral edema,and high-altitude pulmonary edema. Histamine H3 receptor, primarily as a presynaptic receptor, is widely expressed in the central and peripheral systems. Histamine,dopamine,acetylcholine and many other neurotransmitters are regulated by histamine H3 receptor. Studies have shown that histamine H3 receptor is involved in the hypoxic response of the respiratory network. In addition,histamine,espe?cially histamine H3 receptor,participates in the regulation of cerebral ischemia in the central nervous system. In this paper,we reviewed the structure and functions of histamine H3 receptor and explained its role in the regulation of hypoxia so as to evaluate the possibility of histamine H3 receptor as a drug target for the therapy of hypoxia-induced injuries.
7.Antinociception of omega-conotoxin ω-SO3 in rat formalin test after acute or chronic intrathecal administration
Xiaoli JI ; Lingdi YAN ; Peilan ZHOU ; Huajin DONG ; Zehui GONG ; Peitang HUANG
Chinese Pharmacological Bulletin 2010;26(4):476-482
Aim To evaluate the antinociception of ω-SO3,a novel Omega-superfamily conotoxin,in rat formalin test.Methods Potency and duration of ω-SO3 antinociception compared with morphine were investigated in rat formalin test after acute intrathecal administration.Development of tolerance or cross tolerance to analgesia of ω-SO3 and morphine was tested in formalin test after chronic intrathecal administration.Locomotor activity of rat after acute intrathecal administration was tested to evaluate possible central side effects.Results In rat formalin test after intrathecal bolus injection,ω-SO3 produced dose-and time-dependent antinociception by suppressing acute(ED_(50),1.79 ng·g~(-1))and tonic phases(ED_(50),0.41 ng·g~(-1)),which was approximately 10-fold potency and twice longer-acting of morphine in blocking tonic phase responses.After repeated intrathecal injections twice daily for 5 consecutive days,ω-SO3 produced analgesia without loss of potency whereas morphine produced analgesia tolerance in rat formalin test;further,ω-SO3 still produced potent analgesia in morphine-tolerant rats.No changes in motor function were seen in rats receiving above antinociceptive doses.Conclusion sNovel ω-SO3 produces potent and long-acting spinal antinociception without observable motor dysfunction and after chronic intrathecal administration.ω-SO3 produces neither tolerance nor cross-tolerance to morphine analgesia.
8.Effect of inhaled low density nitric oxyde in pig' s pulmonary thromboembolism
Sugang GONG ; Jinming LIU ; Tong ZHOU ; Bo SUN ; Peilan GAO ; Wenzeng LIU ; Xia LI ; Tao GUI
Chinese Journal of Emergency Medicine 2009;18(8):846-850
Objective To investigate the mechanism and significance of low concentration nitric oxide (NO) inhalation in the treatment of pulmonary thromboembelism in swine. Method The pulmonary thromboem-bolism(PTE) model was made in 15 healthy infantile swines which were subsequently assigned to either control group (n = 8) or NO group (n = 7). Swines of the control group were not treated with any medicine, while 10 ppm of NO was administered by continuous inhalation for 2 hours to swines of NO group. Volume of physiological dead space (VDphy), volume of alveolar dead space (VDalv), intrapulmonary shunt (Qs/Qt), mean pulmonary arterial pressure (PAP), systolic blood pressure (SBP), heart rate (HR), cardiac output (CO), arterial blood pH (pH), arterial partial pressure of carbon dioxide (PaCO2) and arterial partial pressure of oxygen (PaO2) were measured 30 min before and 0 min, 30 min, 60 min, 120 min and 180 min after establishment of VIE. Results The post-FIE VDphy, VDalv, Qs/Qt and PAP in both groups increased markedly after PTE compared with the cor-responding pre-PTE measurements (P < 0.01). Post-FIE PaO2 of both groups decreased significandy (P <0.05 and P <0.01), while significance difference was found between pre- and post-PTE HR, SBP, CO, pH or PaCO2 in neither groups (P > 0.05). Both post-PTE PAP and VDalv in NO group were markedly lower(P <0.05 and P <0.01) and beth PaCO2 and PaO2 were much higher than those of the control group (P <0.05). No signi-fieant difference were found in other measurements between two groups. Conclusions Pulmonary arterial pressure may be lowered, alveoli dead space may be reduced and PaCO2 increased by low concentration NO inhalation for the treatment of PIE without decline in haemodynamic status.
9.Establishment of recombinant cell line stably expressing humanα2A-adrenoceptor
Yi YANG ; Yulei LI ; Meng LIU ; Peilan ZHOU ; Ruibin SU ; Zehui GONG
Chinese Journal of Pharmacology and Toxicology 2016;30(5):576-581
OBJECTIVE To establish a new cell line that can stably express humanα2A-adrenoceptor (α2A- AR). METHODS Recombinant plasmid of α2A- AR with hygromycin B (Hygro) resistance (pcDNA3.1/Hygro-HA-α2A-AR)was stably transfected into Chinese hamster ovary(CHO)cells which had expressed protein kinase A catalytic subunits(PKAcat) with labeling of enhanced green fluorescent protein(EGFP)by a Lipofectamine based method. A single positive clone expressingα2A-AR was selected through cultivation in the presence of 200 mg · L-1 hygromycin B followed by PKA redistribution assay. The transcriptional expression ofα2A-AR was detected by quantitative real-time PCR(qRT-PCR). Time-resolved fluorescence resonance energy transfer immunoassay was used to identify the function of inhibiting cAMP accumulation of α2A-AR. RESULTS The CHO-PKAcat-α2A-AR cell line No.7 exhibited stable response in PKA redistribution assay. qRT-PCR analysis demonstrated that the high expression ofα2A-AR in the cell line remained stable after a few generations compared with CHO-PKAcat-EGFP cells (P<0.01). The cAMP accumulation caused by forskolin was significantly inhibited by α2A-AR agonist in CHO-PKAcat-α2A-AR cells(P<0.01). CONCLUSION CHO-PKAcat-α2A-AR cell line is constructed successfully, which provides an effective model for drug screening and studies of mechanisms.
10.Tacrolimus inhibits podocyte injury and stabilizes the expression of cytoskeleton and nephrin
Yueqiang WEN ; Huiyuan LI ; Peilan ZHOU ; Zebin WANG ; Jianbo LIANG
The Journal of Practical Medicine 2017;33(22):3694-3697
Objective To investigate the function and molecular mechanism of tacrolimus in podocyte injury and restoration.Methods Cultured podocytes were stimulated by Angiotensin Ⅱ (Ang Ⅱ) or Ang Ⅱ plus tacrolimus.Cells were collected at different time points (0 h,12 h and 24 h).The distribution of F-actin was observed after immunofluorescence staining,and the protein expression of nephrin and podocin were detected by Western Blot (WB).Results In normal control podocytes,F-actin was arranged in cytoplasm powerfully.Ang Ⅱ induced the disruption and discontinuity of F-actin.Tacrolimus inhibited the effect of Ang Ⅱ,stabilized the regular arrangement the F-actin.Compared to normal cells,the protein expression of nephrin in Ang Ⅱ group significantly decreased at 24 h after stimulation (0.76 ± 0.32 in AngⅡ group vs.1.18 ± 0.40 in normal group,P < 0.05).And tacrolimus stabilized the expression of nephrin protein (1.00 ± 0.19 in treatment group vs.0.76 ± 0.32 in Ang Ⅱ group,P < 0.05).Ang Ⅱ and tacrolimus did not affect the expression of podocin protein.Conclusion Tacrolimus inhibits podocyte injury induced by Ang Ⅱ,stabilizes the regular arrangement of cytoskeleton and protein expression of nephrin.