Application Of SiRNA To Inhibit MOR In The Treatment Of Opioid Addiction

MicroRNAs(miRNAs)are small(19-23 nucleotides)non-coding RNA molecules that have different expression pattern under physiological and pathological conditions resulting in amplification or depression of target gene.For many important oncogene or tumor suppressor gene were regulated by miRNA,miRNA-gene regulation pathways may well explain the development of cancer and provide the potential treatment.siRNAis another double-stranded small RNA(21-25nt)molecule which triggers the degradation of target mRNA by fully complementary pairing with target genes.siRNA is widely used in gene function and therapeutic drug because of its high suppression efficiency,high specificity,no drug toxicity and good tolerance.Opioid addiction is a chronic disease that affects the public health crisis.It not only causes patients to have physical dependence,but also produces strong mental dependence.Once the addiction occurs,it will be difficult to cure,and it is easy to relapse.The damage can be said to be devastating.Opioid addiction is associated with very specific health risks,including the spread of various blood-borne viruses such as human immunodeficiency virus(HTV)and hepatitis B and hepatitis C.In addition,opioid addiction is often linked to violence,crime,AIDS,infectious diseases,mental illness(anxiety,depression,etc.).Opioidreceptors are receptors that mediate endogenous opioid peptides and opioids,mainly including mu receptors,κ receptors,and δ.Among them,the most closely involved in the drug-dependent addiction process is the μreceptor.Most of the drugs currently used to treat addiction are targets of the μ receptor.In this study,MOR protein was used as a research object,and the effect of MOR expression level inhibition on the reward mechanism was studied.First,a specific siRNA was designed for the MOR sequence,and a sequence with the highest inhibition efficiency was screened in vitro,and the siRNA expression plasmid was constructed Since the μ receptor is a central nervous system receptor and drug-targeted therapy needs to cross the blood-brain barrier,we have engineered the constructed siRNA expression plasmid to use the rabies virus glycoprotein RVG and the membrane surface fusion protein RVG-Lamp2b.The sequence in which the glycine chains are ligated together to form a targeting tag is inserted downstream of the CMVpromoter,allowing it to be expressed simultaneously with the siRNA to inhibit expression of the nervous system MOR.We used morphine to induce conditional preference in mice,and injected them with physiological saline,nc siRNA/RVG plasmid,MOR siRNA plasmid,MOR siRNA/RVG plasmid,and then CPP to detect the effect of drug on mouse condition preference.The results showed that the position preference of the mice didnot change much before and after the injection of siRNA,and remained consistent with the spontaneous detoxification results after detoxification before administration.However,once again,mice were injected with morphine once,and the group injected with MOR siRNA/RVG plasmid was significantly shorter in the white box than the other three groups.Combined with the western results of brain tissue,it was shown that the MOR siRNA/RVG plasmid was successfully expressed in vivo to the nervous system,and siRNA interference inhibited MOR expression,which had a certain effect on reducing the body’s desire for relapse.Considering that morphine withdrawal process is often accompanied by negative emotions such as depression,this study also uses three classical depression research experiments to explore the association between addiction treatment process and depressive symptoms.The results showthat there is no necessary correlation between them.In summary,this topic is to develop siRNA/miRNA drugs for MOR,and to find a delivery method that is safe,efficient,and can target the transport of siRNA into central nervous tissues.The perfection of the delivery system is of great significance for the clinical transformation of siRNAtherapy,and the delivery of the engineered plasmid to the endogenous exosome may help siRNA therapy move forward.