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The Analgesic Study of Morphine Tolerance by Down-regulating δ Opioid Receptor in Rats

Author: ZuoKunFang
Tutor: YangHui
School: Huazhong University of Science and Technology
Course: Anesthesiology
Keywords: Lentiviral vectors Morphine tolerance Opioid receptors SNI rats RNAinterference
CLC: R614
Type: Master's thesis
Year: 2013
Downloads: 3
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ObjectiveThe aim of the study was to investigate the effect of δ opioid receptor(DOR)inmorphine tolerance(MT)and to provide a theoretical basis for finding a reasonable way ofanalgesic through the research of chronic morphine analgesic tolerance rats withneuropathic pain.MethodsA lentiviral-mediated short hairpin RNA (shRNA) system was synthesized to deliverthe DOR shRNAs to the spinal cord of the SNI rats.10days after transfection usingreal-time PCR to determine the expression level of DOR mRNA so as to confirm the effectof transfection of the lentivirus. MT was induced by intrathecal morphine (10μg b.i.d) forconsecutive11days. Mechanical paw withdrawal threshold were assessed to determine theanalgesic effects of morphine. Expression of DOR mRNA and protein was determined byreal-time polymerase chain reaction and western blotting analysis immunohistochemistyrespectively.Results10days after siRNA injection, the expression of DOR mRNA was significantlydown-regulate in the spinal cord of group which were injected with LV-shDOR comparedto the rats in group without silencing siRNA. Compared to the control group, a single injection of LV-shDOR significantly preventedmorphine antinociceptive tolerance and shows a stable analgesic effect. DOR mRNA andprotein levels were dramatically down-regulated in the LV-shDOR group and differently,chronic administration of morphine profoundly decreased the level of MOR inmorphine-tolerant rats. The expression of MOR was not reduced in the spinal cordcompared to the rats without silencing siRNA. The chronic administration of morphineinduced a stabilized analgesic tolerance.ConclusionsThis study have proved that a single injection of LV-shDOR can successfullydown-regulate DOR in the spinal cord and prevent the development of morphine tolerance.In addition,long time use of morphine intrathecally can cause the up-regulation of DORand down-regulation of MOR in the spinal cord. DOR play an important role in morphinetolerance.

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