左归降糖通脉方对2型糖尿病合并脑梗死大鼠坏死性凋亡通路的影响  

Effects of Zuogui Jiangtang Tongmai Recipe on necroptosis pathway in a rat model of type 2 diabetes mellitus complicated with cerebral infarction

作　　者：蔡昱哲 李定祥 刘艺璇 罗政 阳晶晶 雷翰霖[4] 张亚男 吴琴 陈静 邓奕辉[2,3] CAI Yu-zhe;LI Ding-xiang;LIU Yi-xuan;LUO Zheng;YANG Jing-jing;LEI Han-lin;ZHANG Ya-nan;WU Qin;CHEN Jing;DENG Yi-hui(College of Integrated Traditional Chinese and Western Medicine,Hunan University of Chinese Medicine,Changsha 410208,China;Hunan Provincial Key Laboratory of Integrated Traditional Chinese and Western Medicine for the Prevention and Treatment of Cardio-Cerebral Diseases,Hunan University of Chinese Medicine,Changsha 410208,China;School of Traditional Chinese Medicine,Hunan University of Chinese Medicine,Changsha 410208,China;The First Hospital Affiliated to Hunan University of Chinese Medicine,Changsha 410007,China)

机构地区：[1]湖南中医药大学中西医结合学院,湖南长沙410208 [2]湖南中医药大学中西医结合心脑疾病防治湖南省重点实验室,湖南长沙410208 [3]湖南中医药大学中医学院,湖南长沙410208 [4]湖南中医药大学第一附属医院,湖南长沙410007

出　　处：《中成药》2024年第9期2936-2942,共7页Chinese Traditional Patent Medicine

基　　金：湖南省科技创新团队项目(2020RC4050);湖南省教育厅科学研究项目(21A0228);湖南省中医药科研计划项目(E2022010);校级研究生创新课题(2022CX11);2021年度中西医结合一流学科重点项目(2021ZXYJH08)。

摘　　要：目的探讨左归降糖通脉方对2型糖尿病(T2DM)并发脑梗死(CI)大鼠坏死性凋亡通路的影响。方法将SD大鼠随机分为假手术组、模型组、二甲双胍组(0.045 g/kg)和左归降糖通脉方低、中、高剂量组(6.5、13、26 g/kg),每组9只。除假手术组外,其余各组大鼠通过喂养高糖高脂饲料4周结合腹腔注射链脲佐菌素建立T2DM大鼠模型,血糖稳定1周后,进行大脑中动脉阻塞(MCAO)造模,并在MCAO术前3 d开始灌胃给予相应剂量药物。给药7 d后,采用mNSS法、TTC染色评价脑梗死损伤效应;采用血糖仪检测血糖;ELISA法检测糖化血清蛋白水平和血清TNF-α、IL-1β水平;RT-qPCR法检测脑组织FADD、RIPK1、RIPK3、MLKL mRNA表达;Western blot法检测脑组织FADD、p-RIPK1、p-RIPK3、p-MLKL蛋白表达。结果与假手术组比较,模型组血糖值、糖化血清蛋白、神经功能评分、脑梗死体积、脑组织FADD、RIPK1、RIPK 3、MLKL mRNA表达、脑组织FADD、p-RIPK1、p-RIPK3、p-MLKL蛋白表达、血清TNF-α、IL-1β水平均升高(P<0.01),神经元排列紊乱、形态多样、胞核缩小;与模型组比较,左归降糖通脉方中、高剂量组及二甲双胍组以上指标均有改善(P<0.05,P<0.01),且神经元形态较规则、排列较整齐、细胞间隙缩小。结论左归降糖通脉方能够改善T2DM并发CI大鼠的神经功能障碍,其作用可能与抑制坏死性凋亡信号通路的激活有关。AIM To investigate the effects of Zuogui Jiangtang Tongmai Recipe on necroptosis pathway in a rat model of type 2 diabetes mellitus(T2DM)complicated with cerebral infarction(CI).METHODS The SD rats were randomly divided into the sham operation group,the model group,the metformin group(0.045 g/kg),and the low,medium and high dose Zuogui Jiangtang Tongmai Recipe groups(6.5,13,26 g/kg),with 9 rats in each group.In contrast to rats of the sham operation group,rats of the other groups were given 4 weeks feeding of high-sugar and high-fat diet combined with intraperitoneal injection of streptozotocin to establish a T2DM rat model with one week stable blood glucose,followed by gavage of corresponding drugs 3 days before the establishment of the middle cerebral artery occlusion(MCAO)model.After 7 days of administration,the rats had their CI injury assessed by mNSS method and TTC staining;their level of blood glucose detected by blood glucose meter;their levels of glycated serum protein,serum TNF-αand IL-1βdetected by ELISA;their cerebral mRNA expressions of FADD,RIPK1,RIPK3 and MLKL detected by RT-qPCR;and their cerebral protein expressions of FADD,p-RIPK1,p-RIPK3 and p-MLKL detected by Western blot.RESULTS Compared with the sham operation group,the model group displayed increased levels of blood glucose value,glycosylated serum protein,neurological function score,cerebral infarction volume,cerebral FADD,RIPK1,RIPK3 and MLKL mRNA expressions,cerebral FADD,p-RIPK1,p-RIPK3 and p-MLKL protein expressions,serum TNF-αand IL-1βlevels(P<0.01);and more disordered and morphologically diverse neurons with smaller nucleus.Compared with the model group,the groups intervened with medium or high dose Zuogui Jiangtang Tongmai Recipe,or metformin shared improvement in terms of the aforementioned indices(P<0.05,P<0.01);and more neurons with regular morphology neat arrangement,and reduced cell gap.CONCLUSION Zuogui Jiangtang Tongmai Recipe can improve the neurological dysfunction of the rat model of T2DM complicated with CI,which may

关 键 词：左归降糖通脉方 2型糖尿病 脑梗死 坏死性凋亡 

分 类 号：R285.5[医药卫生—中药学]