糖尿病抑制局麻药致神经元DNA损伤后重组修复通路中DNA-PK的机制研究

Patients with diabetes in the accept for nerve block anesthesia or analgesic therapy , the risk of nerve injury was significantly increased. Our research team previous study showed: under high glucose conditions local anesthetic (LA)after DNA damage induced nerve cell repair enzyme XPD decreased expression,neuronal apotosis was significantly increased.Inferred:high glucose increased LA neurotoxicity mechanism of inhibition is unknown?The repair pathways after of LA induced nerve cell damage are not reported. We speculate that high glucose increased local anesthetic neurotoxicity is related to that high glucose inhibits nerve cell repair. Our pre-experiments about the screening repair genes showed the DNA-PK which is one gene of the recombination repair pathway was significantly increased,while in high glucose it was significantly inhibited. In this study the cDNA microplate array technology and various molecular biology methods will be used , through the high expression and low expression gene DNA-PK,BRCA1,ATM cell line in vitro , and through the knockout and overexpression of the IG in vivo experiments,to investigate the the exact mechanism of nerve cell DNA damage repair caused by local anesthetics, and the high glucose environment DNA damage repair pathway inhibited reasons. This study provides new ideas and targets for elucidating the mechanisms of local anesthetic induced neural cells DNA damage repair and further for the prevention and treatment of diabetic peripheral nerve complications

糖尿病患者在行神经阻滞麻醉或镇痛治疗后，神经损伤的风险明显增加。本科研小组前期研究表明：在高糖环境下局麻药（LA）致神经细胞DNA损伤后XPD修复酶表达减少，神经细胞凋亡明显增多。由此推测：高糖环境加重LA神经毒性作用机制与其抑制DNA损伤修复通路有关,但其机制不明？LA致神经细胞DNA损伤后何修复通路参与也未见报道。我们行修复基因筛查的预实验显示LA致神经细胞DNA损伤后重组修复通路中DNA-PK表达显著增高，而高糖环境下其明显抑制。为进一步研究重组修复通路在LA致神经细胞DNA损伤后的作用及高糖抑制该修复通路的机制，本研究拟采用cDNA微孔板列阵技术和多种分子生物学方法，通过构建基因筛查出表达明显差异的重组修复通路基因DNA-PK、BRCA1、ATM等高表达和低表达细胞株；并在敲除和过表达上述基因的小鼠上予以验证，为LA神经损伤及糖尿病加重其损伤的防治提供新的思路和靶点。

糖尿病患者在行神经阻滞麻醉或镇痛治疗后，神经损伤的风险明显增加。本科研小组前期研究表明：在高糖环境下局麻药致神经细胞 DNA 损伤后 XPD 修复酶表达减少，神经细胞凋亡明显增多。由此推测：高糖环境加重局麻药神经毒性作用机制与其抑制 DNA损伤修复通路有关,但其机制不明？局麻药致神经细胞 DNA 损伤后何修复通路参与也未见报道。我们行修复基因筛查的预实验显示布比卡因致神经细胞 DNA 损伤后重组修复通路中DNA-PK复合物 表达显著增高，而高糖环境下其明显抑制。. 为进一步研究重组修复通路在局麻药致神经细胞DNA损伤后的作用及高糖抑制该修复通路的机制，我们构建了相应目标修复基因（Ku70）的高表达细胞株，并在离体和在体糖尿病模型上进一步研究其在局麻药及高糖环境下对神经细胞的修复作用；验证高糖为何加重布比卡因引发神经损伤机制。为布比卡因导致神经损伤及糖尿病加重其损伤的防治提供新的思路和靶点。