基于RIPK1/RIPK3/MLKL途径探讨温肺降浊方对VaD大鼠坏死性凋亡的影响  

The Wenfei Jiangzhuo Formula exerts protective effects on VaD model rats by inhibiting nec⁃rotic apoptosis through attenuating the RIPK1/RIPK3/MLKL pathway

作　　者：张鼎 陈炜[2] 秦红玲[2] 张玲[1] 向军军[2] 王开龙[2] 姚春[1] 胡跃强[2] ZHANG Ding;CHEN Wei;QING Hong-ling;ZHANG Ling;XIANG Jun-jun;WANG Kai-long;YAO Chun;HU Yue-qiang(Guangxi University of Chinese Medicine,Nanning,Guangxi 530001,China;The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning,Guangxi 530001,China)

机构地区：[1]广西中医药大学,广西南宁530001 [2]广西中医药大学第一附属医院,广西南宁530023

出　　处：《时珍国医国药》2025年第1期1-8,共8页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(82260904);广西中医药大学“桂派杏林拔尖人才”计划(04B2300807,04B2300907);国家中医药管理局高水平重点学科-中医内科学(ZYYZDXK-2023166);广西中医药大学“歧黄工程”高层次人才团队(202410);广西中医药重点学科建设项目(GZXK-Z-20-13)。

摘　　要：目的探讨温肺降浊方是否抑制RIPK1/RIPK3/MLKL途径减少坏死性凋亡,从而对血管性痴呆(VD)模型大鼠发挥脑保护作用。方法健康雄性SD大鼠随机分为假手术组、模型组、抑制剂组、温肺降浊方低剂量、中剂量和高剂量组,除假手术组,其余各组制备VaD模型,造模成功后各组以相应药物灌胃,治疗3周后进行各项检测。观察各组大鼠行为学、海马CA1区形态学损伤和Nissl小体变化情况,ELISA检测血清中IL-1β、IL-6和TNF-α浓度;Western blot和qRT-PCR检测RIPK1/RIPK3/MLKL通路蛋白及mRNA的表达情况。结果与假手术组比较,模型组和各干预组大鼠逃避潜伏期延长(P<0.05),穿越平台次数缩短(P<0.05),海马神经元形态稀松,部分细胞形态破裂,出现水肿,细胞核缩小,Nissl小体严重缺失,结构空洞;血清中IL-1β、IL-6和TNF-α表达升高(P<0.05);大鼠海马组织中RIPK1/RIPK3/MLKL通路蛋白及mRNA表达升高(P<0.05);与模型组比较,抑制剂组和温肺降浊方高剂量组大鼠逃避潜伏期缩短(P<0.05),穿越平台次数增加(P<0.05),海马神经元数量增多,细胞排列紧密,形态逐渐规整,结构较为完整;血清中IL-1β、IL-6和TNF-α表达下降(P<0.05);海马组织中RIPK1/RIPK3/MLKL通路蛋白及mRNA表达下降(P<0.05)。结论温肺降浊方可以抑制RIPK1/RIPK3/MLKL途径以减少坏死性凋亡,从而改善大脑认知情况,发挥脑保护效应。Objective To investigate whether the Wenfei Jiangzhuo Formula inhibits RIPK1/RIPK3/MLKL pathway to reduce necrotic apoptosis and thus exerts cerebroprotective effects on VaD model rats.Methods Healthy male SD rats were randomly divided into the Sham group,the model group,the inhibitor group,and the low-dose,medium-dose and high-dose groups of the Wenfei Jiangzhuo Formula;except for the Sham group,the other groups were prepared as VaD models,and each group was gavaged with the corresponding drugs after successful modeling,and the tests were carried out after 3 weeks of treatment.Behavior,morphological damage in hippocampal CA1 area and changes in Nissl vesicles were observed in each group.Serum concentrations of IL-1β、IL-6 and TNF-α were detected by ELISA;and the expression of RIPK1/RIPK3/MLKL pathway proteins and mRNAs were detected by Western blot and qRT-PCR.Results Compared with the Sham group,rats in the model group and each intervention group had prolonged evasion latency(P<0.05)and shortened number of crossing platforms(P<0.05);hippocampal neurons had a sparse morphology,with ruptured morphology of some cells,edema,shrunken nuclei,and severe deletion of Nissl vesicles with structural vacuoles;and the expression of IL-1β,IL-6,and TNF-α in the serum was elevated(P<0.05);The expression of RIPK1/RIPK3/MLKL pathway proteins and mRNA in rat hippocampal tissues was elevated(P<0.05);compared with the model group,the evasion latency of rats in the inhibitor group and the high-dose group of the Wenfei Jiangzhuo Formula was shortened(P<0.05),and the number of traversals across the plateau was increased(P<0.05);the number of hippocampal neurons increased,the cells were arranged tightly,and their morphology gradually regularized,and their structure more intact;the expression of IL-1β,IL-6 and TNF-α in serum decreased(P<0.05);the expression of RIPK1/RIPK3/MLKL pathway protein and mRNA in hippocampal tissue decreased(P<0.05).Conclusion The Wenfei Jiangzhuo Formula can inhibit the RIPK1/RIPK3/MLKL pathway to reduce

关 键 词：温肺降浊方 坏死性凋亡 RIPK1/RIPK3/MLKL信号通路 血管性痴呆 神经元凋亡 作用机制 

分 类 号：R285.5[医药卫生—中药学]