| Opioids were widely applied to treatment of analgesia and treatment against coughing, diarrhea, anxiety and sleepless. Opioids had toxic effects while playing pharmaceutical roles; Oxidative injury was one of the most important toxic effects. Multi-organ or systems in our body were more liable to be damaged either by opioids induced reduction in concentration of glutathione, the reductant or antioxidant or by opioids induced increase in concentration of reactive oxidative species. To further clarify the research progress in opioids induced oxidative injury is very important towards opioids applications as well as prevention and cure of drug abuse. NO is synthesized from L-arginine by the endothelial NO synthase (eNOS), together with a stoechionetric production of L-citrulline. The reaction requires molecular oxygen. NO is affinitive with the pathophysiologic progress in kidney. It is getting more and more attribute importance to the relationship between morphine dependence and NOS. Nitric oxide synthase (NOS) is the key enzyme to catalystic and synthesize NO in vivo. NOS was documented with three isoforms, the neuronal NOS (nNOS), the inducible (iNOS), and the endothelial NOS (eNOS). Nitric oxide synthase (NOS) inhibitors attenuate or block morphine action. NO/NOS system has an affect on morphine-induced place preference. Action of NO/NOS system are involved in the activation of opioid receptors. The effect of NO/NOS system on the expression of morphine withdrawal symptoms may be partially mediated by noradrenergic neuron system. The aquaporins are a family of small, membrane-spanning proteins the act as highly selective water channels. They were found as the special channel existing in the cell membrane of mammalian and vegetation. AQP-1 was the first water channel protein that was cloned in erythrocyte cell membrane, its molecular weight is 28kD. AQP-1 distributes extensively on the endodermis cell of capillary vessel in many tissues, such as kidney, lung and airway, osseous muscles, peritoneum, pleura, glands. They also located in choroids coat epithelial cell in brain, corneal, small intestines, placenta and chorionic epithelium. AQP-1 is present in the cells of the proximal tubule and the descending limb of Henle's hoop and endodermis cell in exterior medulla. Opioid strongly affect the renal function by variant mechanism. Both AQP-1 and eNOS were expressed in the endodermis cells in exterior medulla. We investigated the relationship between morphine induced renal injury and the changes of eNOS and AQP-1 gene expression in kidney of morphine dependent rat. The main contents of this paper are the following: Part one: Establishing the morphine dependent rat model: We established the morphine dependent rats model successfully through administrating morphine by intraperitoneal injection twice per day. Rats represent dependence and withdraw syndromes along with the administrating time passing by. The weight of natural withdraw rats is descending along with the administrating time passing by. Part two: Identify the RT-PCR amplified eNOS product by restriction enzyme. We apply restriction enzyme to identify if the eNOS RT-PCR product is our objective gene with the help of bioinformatic, biology software and internet. The RT-PCR product was verified preliminarily being aim eNOSgene. The eNOS bands was amplified in cardiac and purified with PCR fragment Recovery Kit and digested with BamHâ… , the resultant product conformed the anticipated consequence, that is the RT-PCR product should be eNOS gene fragment. Part Three: The pathological histology of kidney in morphine dependent rats. A physical dependent model in rats was established by intraperitoneal injection of morphine and the pathological and histological changes in kidney tissue were reviewed by light microscope. In the natural withdraw rats, hydropic change and degeneration was found in kidney cells of all morphine dependent rats. It was also seen heterogeneous vacuolar degeneration, interstitial angiectasis, glomerulus pycnosis and necrosis. Thus there is mild pathological damage in kidney tissue of morphine dependent rats. Part four: Changes of eNOS gene expression in kidney of morphine dependent rats. Changes of NO content was determined the level of NOS mRNA. The result of our study showed the changes of eNOS mRNA expression in morphine dependent rats was very similar with other document. During the chronic progress, after different administrating time, the eNOS mRNA content was detected by RT-PCR, southern blot hybridization. Comparing with the control group, the content of eNOS gene transcription changed unapparently after administration of morphine for 3 days, and tended to increase after administration for 7 days, raised significantly after administration for 7 days then stopped for 3days. The increasing trend was a little descending after stopping administration for 7days, but still kept in higher level. The level of eNOS in kidney of morphine dependent rats showed different changes at varied degree. But the expression of AQP-1 was very...
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