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为观察导入CTLA4Ig基因能否延长胰岛素基因治疗的疗效时间,静脉注射链脲佐菌素(streptozotocin,STZ)制备1型糖尿病(type 1diabetes mellitus,T1DM)小鼠模型,将24只造模成功的雄性C57小鼠随机分为注射空载腺病毒组、表达胰岛素的腺病毒组(recombinant adenovirus vector/insulin,rAdV/Ins)和共表达胰岛素和CTLA4Ig基因的重组腺病毒(recombinant adenovirus vector/insulin-CTLA4Ig,rAdV/Ins-CTLA4Ig)组,每组8只。采用实时定量聚合酶链反应(quantitative real-time polymerase chain reaction,qRT-PCR)方法检测胰岛素、CTLA4Ig及其相关基因在小鼠肝组织的mRNA表达水平。应用简便血糖仪定期检测小鼠血糖水平。rAdV/Ins-CTLA4Ig组小鼠肝组织中CTLA4Ig mRNA的表达量显著高于rAdV/Ins组和空载腺病毒组(P<0.01)。rAdV/Ins-CTLA4Ig组小鼠肝组织中IFN-γmRNA的表达显著低于空载腺病毒组(P=0.04)。与注射空载腺病毒组相比较,rAdV/Ins-CTLA4Ig和rAdV/Ins两组小鼠的血糖水平显著降低(均P<0.05),且rAdV/Ins-CTLA4Ig组小鼠较rAdV/Ins组小鼠能维持更长的正常血糖时间(P<0.01)。胰岛素和CTLA4Ig共表达基因的重组腺病毒能转染小鼠肝脏,CTLA4Ig能在肝内成功表达并通过抑制炎症反应延长了胰岛素基因表达时间。
To observe whether the introduction of CTLA4Ig gene can prolong the therapeutic effect of insulin gene therapy, a mouse model of type 1 diabetes mellitus (T1DM) was established by intravenous injection of streptozotocin (STZ) C57 mice were randomly divided into the group of injected empty adenovirus, recombinant adenovirus vector (rAdV / Ins) and recombinant adenovirus vector / insulin-CTLA4Ig co-expressing insulin and CTLA4Ig gene, rAdV / Ins-CTLA4Ig) group, 8 in each. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect mRNA expression of insulin, CTLA4Ig and related genes in mouse liver. Blood sugar level in mice was measured regularly by simple blood glucose meter. The expression of CTLA4Ig mRNA in rAdV / Ins-CTLA4Ig group was significantly higher than that in rAdV / Ins group and blank adenovirus group (P <0.01). The expression of IFN-γmRNA in liver tissue of rAdV / Ins-CTLA4Ig group was significantly lower than that of empty adenovirus group (P = 0.04). Compared with the rAdV / Ins group, the rAdV / Ins-CTLA4Ig and rAdV / Ins groups had significantly lower blood glucose levels (all P <0.05), compared with the rAdV / Ins group Rats maintained longer normal blood glucose time (P <0.01). Recombinant adenovirus expressing both insulin and CTLA4Ig could transfect mouse liver. CTLA4Ig could be successfully expressed in the liver and prolonged insulin gene expression by inhibiting the inflammatory response.