基于自噬探讨左归降糖通脉方对糖尿病合并脑梗死大鼠的神经保护作用机制  

Exploring the Mechanism of Neuroprotective Effect of Zuogui Jiangtang Tongmai Prescription on Diabetic Rats with Cerebral Infarction Based on Autophagy

作　　者：李钰佳 彭珣 李定祥[1] 邓奕辉[1] LI Yujia;PENG Xun;LI Dingxiang;DENG Yihui(Hunan University of Chinese Medicine,Changsha 410208 Hunan,China)

机构地区：[1]湖南中医药大学,湖南长沙410208

出　　处：《中药新药与临床药理》2023年第2期176-183,共8页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金面上项目(81874416);湖南省科技创新团队项目(2020RC4050);湖南省中医药科研计划项目(E2022010);湖南省教育厅科研项目(21A0228,20A365)。

摘　　要：目的 从自噬角度探讨左归降糖通脉方(熟地黄、枸杞子、山茱萸、黄芪、黄连等)对糖尿病合并脑梗死大鼠的神经保护作用机制。方法 将96只雄性SD大鼠随机分为假手术组、模型组、左归降糖通脉方组(24 g·kg^(-1))及二甲双胍+阿司匹林组(150 mg·kg^(-1)+15 mg·kg^(-1)),每组24只。除假手术组大鼠外,其余大鼠采用高糖高脂饲料喂养4周后腹腔注射链脲佐菌素建立2型糖尿病大鼠模型,模型复制成功后开始灌胃给药,每日1次,连续5 d。末次灌胃后采用线栓法制备大脑中动脉栓塞大鼠模型。术后进行神经功能评分;采用TTC染色法测定脑梗死率;检测大鼠血糖值及糖化血清蛋白含量;HE染色观察缺血半脑皮质区组织损伤;ELISA法检测缺血侧大脑皮质组织中白细胞介素1β(IL-1β)、IL-6、肿瘤坏死因子α(TNF-α)的含量;Western Blot法检测皮质组织中微管相关蛋白轻链3(LC3)、p62蛋白表达情况;免疫荧光法检测皮质组织中LC3蛋白表达。结果 与假手术组比较,模型组大鼠的神经功能损伤评分、脑梗死率、血糖水平、治疗前后的体质量差值及糖化血清蛋白水平显著上升(P<0.01);脑缺血侧皮质组织中IL-1β、IL-6、TNF-α含量显著升高(P<0.01),LC3-Ⅱ/Ⅰ蛋白表达显著上调(P<0.01),p62蛋白表达显著下调(P<0.01),LC3荧光表达面积显著增加(P<0.01)。与模型组比较,左归降糖通脉方组大鼠的神经功能评分及脑梗死率明显下降(P<0.05),血糖水平、治疗前后的体质量差值及糖化血清蛋白水平均显著降低(P<0.01);皮质组织中IL-1β、IL-6、TNF-α含量显著降低(P<0.01),LC3-Ⅱ/Ⅰ蛋白表达显著下调(P<0.01),p62蛋白表达显著上调(P<0.01),LC3荧光表达面积显著减少(P<0.01)。结论 左归降糖通脉方能降低糖尿病合并脑梗死大鼠的神经功能损伤,缩小脑梗死体积,降低血糖及糖化血清蛋白水平,减轻大鼠缺血侧大脑皮质炎性损伤,其机制与通过抑制大鼠Objective To investigate the neuroprotective mechanism of Zuogui Jiangtang Tongmai Prescription(Rehmanniae Radix praeparata,Lycii Fructus,Corni Fructus,Astragali Radix,Coptidis Rhizoma)in rats with diabetes mellitus complicated with cerebral infarction from the perspective of autophagy.Methods Ninety-six healthy male SD rats were randomly divided into the sham-operation group,the model group,the Zuogui Jiangtang Tongmai Prescription group(24g·kg^(-1))and the Metformin+Aspirin group(150 mg·kg^(-1)+15 mg·kg^(-1)),with 24 rats in each group.Except for the sham-operation rats,the rats were fed with high sugar and high fat diet for 4 weeks and then injected intraperitoneally with streptozotocin to establish a type 2 diabetic rat model.After successful replication of the model,intragastric administration was started once daily for concecutive 5 days.A rat middle cerebral artery embolism model was prepared by the suture-occluded method after the final gavage.The neurological function score was performed after the operation;the cerebral infarction rate was determined by TTC staining;the tissue damage in the ischemic hemispheric cortical area was observed by HE staining;the contents of interleukin 1β(IL-1β),IL-6 and tumour necrosis factor α(TNF-α)in the cortical tissue on the ischemic side were measured by ELISA;the protein expressions of microtubule-associated protein light chain 3(LC3)and p62 in cortical tissue were detected by Western Blot;the protein expression of LC3 in rat cortex was detected by immunofluorescence method.Results Compared with the sham-operation group,the neurological impairment scores,cerebral infarction rates,blood glucose levels,body mass differences before and after treatment and glycated serum protein level of rats in model group were significantly increased(P<0.01);the levels of IL-1β,IL-6 and TNF-α were significantly increased in cortical tissue on the ischemic side of the brain(P<0.01),LC3-Ⅱ/Ⅰ protein expression was significantly up-regulated(P<0.01),p62 protein expression was si

关 键 词：左归降糖通脉方 糖尿病合并脑梗死 自噬 微管相关蛋白轻链3 P62 大鼠 

分 类 号：R285.5[医药卫生—中药学]