脊髓SGK-1/Kalirin-7/NR2B信号通路在大鼠2型糖尿病神经病理性痛中的作用  

Role of spinal SGK-1/Kalirin-7/NR2B signaling pathway in type 2 diabetic neuropathic pain in rats

作　　者：陈佳丽 张茂表 曹红[1] 李军[1] Chen Jiali;Zhang Maobiao;Cao Hong;Li Jun(Department of Anesthesiology and Perioperative Medicine,the Second Affiliated Hospital&Yuying Children′s Hospital,Wenzhou Medical University,Wenzhou 325024,China)

机构地区：[1]温州医科大学附属第二医院育英儿童医院麻醉与围术期医学科,温州325024

出　　处：《中华麻醉学杂志》2025年第1期65-70,共6页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(81771487);温州市基础性公益科研项目(Y2023972)。

摘　　要：目的评价脊髓血清和糖皮质激素诱导激酶1(SGK-1)/Kalirin-7/含2B亚基的N-甲基-D-天冬氨酸受体(NR2B)在大鼠2型糖尿病神经病理性痛(DNP)中的作用。方法清洁级雄性SD大鼠72只,6周龄,体质量120~160 g,随机接受正常饲料喂养[正常对照组(C组,n=12)]和高脂高糖喂养(n=60)。采用高脂高糖喂养8周后腹腔注射链脲佐菌素(STZ)35 mg/kg的方法制备2型糖尿病模型。以2型糖尿病大鼠痛阈[机械缩足反应阈(MWT)和热缩足潜伏期(TWL)]下降至基础值的85%以下为DNP建模成功。取2型糖尿病建模成功的非DNP(痛阈高于85%基础值)大鼠12只,纳入2型糖尿病非DNP组(NDNP组)。取DNP建模成功的大鼠36只,采用随机数字表法分为3组(n=12):DNP组、DNP+SGK-1抑制剂GSK-650394组(DNP+G组)、DNP+溶剂对照组(DNP+SC组)。于STZ注射后14 d,DNP+G组大鼠鞘内注射300 nmol/L GSK-650394(10μl),每天1次,连续给药14 d;DNP组不做任何处理,DNP+SC组鞘内注射等容量溶剂二甲基亚砜。于喂养8周后、STZ注射后14 d和鞘内注射3、7、14 d(即STZ注射后17、21、28 d)时测定MWT和TWL。于鞘内注射7 d后,取大鼠脊髓腰膨大组织,采用Western blot法测定SGK-1、磷酸化SGK-1(Ser422-SGK1)、Kalirin-7、磷酸化NR2B(Tyr1472-NR2B)和NR2B的表达水平,并计算Ser422-SGK1/SGK1比值和Tyr1472-NR2B/NR2B比值。结果与C组比较,DNP组STZ注射后14 d、鞘内注射相应时点MWT下降,TWL缩短,脊髓Ser422-SGK1/SGK1比值和Tyr1472-NR2B/NR2B比值升高,Kalirin-7表达上调(P<0.05),NDNP组上述指标差异无统计学意义(P>0.05);与DNP组比较,DNP+G组鞘内注射3、7、14 d时MWT升高,TWL延长,脊髓组织Ser422-SGK1/SGK1比值和Tyr1472-NR2B/NR2B比值降低,Kalirin-7表达下调(P<0.05),DNP+SC组上述指标差异无统计学意义(P>0.05)。结论DNP大鼠脊髓SGK-1磷酸化水平增强,可能通过激活Kalirin-7/NR2B信号通路,在DNP的发生及维持中起重要作用。ObjectiveTo evaluate the role of spinal serum and glucocorticoid-inducible kinase 1(SGK-1)/Kalirin-7/N-methyl-D-aspartate receptor 2B(NR2B)signaling pathway in the type 2 diabetic neuropathic pain(DNP)in rats.MethodsClean-grade male Sprague-Dawley rats,aged 6 weeks,weighing 120-160 g,were randomized to receive a normal diet(normal control group[group C,n=12])or a high-fat and high-glucose diet(n=60).Diabetes mellitus was induced by single intraperitoneal injection of streptozotocin 35 mg/kg after 8 weeks of a high-fat and high-glucose diet.The successful DNP model was defined as a decrease in the pain threshold(mechanical paw withdrawal threshold[MWT]and thermal paw withdrawal latency[TWL])to less than 85%of the baseline value in type 2 diabetic rats.Twelve non-DNP(pain threshold above 85%of the baseline value)rats with type 2 diabetes mellitus were randomly selected and included in non-DNP group(NDNP group).Thirty-six rats with DNP were divided into 3 groups(n=12 each)using a random number table method:DNP group,DNP plus SGK1 inhibitor GSK-650394 group(DNP+G group),and DNP plus solvent control group(DNP+SC group).At day 14 after STZ injection,GSK-650394(10μl)300 nmol/L was intrathecally injected once a day for 14 consecutive days in DNP+G group.DNP group did not receive any treatment,and the rats in DNP+SC group received intrathecal injection of the equal volume of dimethyl sulfoxide.The MWT and TWL were measured after 8 weeks of feeding,at 14 days after STZ injection,and at 3,7 and 14 days after intrathecal injection(at 17,21 and 28 days after STZ injection).After 7 days of intrathecal injection,the lumbar enlargement tissues of the spinal cord were collected for determination of the expression of SGK-1,phosphorylated SGK-1(Ser422-SGK1),Kalirin-7,phosphorylated NR2B(Tyr1472-NR2B),and NR2B by Western blot.The ratios of Ser422-SGK1 to SGK1 and Tyr1472-NR2B to NR2B were calculated.ResultsCompared with C group,the MWT was significantly decreased and TWL was shortened at 14 days after STZ injection and at the corre

关 键 词：Rho鸟嘌呤核苷酸交换因子 受体 N-甲基-D-天冬氨酸 糖尿病 2型 糖尿病神经病变 血清和糖皮质激素诱导激酶1 

分 类 号：R614[医药卫生—麻醉学]