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基于PI3K/AKT/mTOR信号通路探讨化瘀通络灸促血管性痴呆大鼠髓鞘再生的作用机制OACSTPCD

中文摘要英文摘要

目的 观察化瘀通络灸对血管性痴呆(vascular dementia,VD)大鼠胼胝体磷脂酰肌醇 3 激酶(phosphatidyli-nositol 3 kinase,PI3K)/蛋白激酶B(protein kinase B,AKT)/哺乳动物雷帕霉素靶蛋白(mammalian target of rapa-mycin,mTOR)信号通路的影响,探讨化瘀通络灸促 VD大鼠髓鞘再生的作用机制.方法 经 Morris水迷宫筛选后,随机选取 12 只大鼠纳入假手术组,剩余大鼠复制 VD模型成功后,随机分为模型组、艾灸组、艾灸+LY294002组,每组 12 只.艾灸组予以化瘀通络灸干预,艾灸+LY294002 组在化瘀通络灸干预的基础上予以 PI3K抑制剂LY294002 腹腔注射,采用 Longa评分法评价各组大鼠神经功能损伤程度,Morris水迷宫实验检测各组大鼠学习记忆能力,Western blot法检测各组大鼠 PI3K/AKT/mTOR信号通路相关蛋白的表达水平,神经髓鞘固蓝染色法观察各组大鼠胼胝体髓鞘的形态,透射电子显微镜观察各组大鼠髓鞘超微结构.结果 与假手术组比较,模型组和艾灸+LY294002 组大鼠的 Longa评分显著升高(P<0.05),逃避潜伏期显著延长(P<0.05),PI3K/AKT/mTOR通路相关蛋白表达水平显著降低(P<0.05),胼胝体内髓鞘纹理不清,排列混乱,边缘呈空泡或空网状改变,髓鞘线圈样结构离散,部分膨出和崩解,有髓神经轴突数量显著减少(P<0.05);与模型组和艾灸+LY294002 组比较,艾灸组大鼠 Longa评分显著下降(P<0.05),逃避潜伏期显著缩短(P<0.05),PI3K/AKT/mTOR通路相关蛋白表达水平显著提高(P<0.05),胼胝体内髓鞘结构有所恢复,排列整齐,边缘结构较为致密,有髓神经轴突数量显著增加(P<0.05).结论 化瘀通络灸可能通过激活 PI3K/AKT/mTOR通路,修复 VD大鼠损伤髓鞘并促进其重塑,恢复脑白质功能.

Objective To investigate the mechanism of action of stasis-resolving and collateral-dredging moxibustion in promo-ting myelin regeneration in rats with vascular dementia(VD)by observing its effect on the phosphoinositide 3 kinase(PI3K)/protein kinase B(AKT)/mammalian target of rapamycin(mTOR)signaling pathway.Methods After screening by the Morris water maze test,12 rats were randomly selected as sham-operation group,and the remaining rats were randomly divided into model group,moxibustion group,and moxibustion+LY294002 group after successful modeling of VD,with 12 rats in each group.The rats in the moxibustion group were given stasis-resolving and collateral-dredging moxibustion,and those in the mox-ibustion+LY294002 group were given intraperitoneal injection of the PI3K inhibitor LY294002 in addition to stasis-resolving and collateral-dredging moxibustion.The Longa scoring system was used to evaluate the degree of neurovascular injury in rats;the Morris water maze test was used to evaluate the learning and memory abilities of rats;Western blot was used to measure the expression of proteins associated with the PI3K/AKT/mTOR signaling pathway;LFB myelin staining was used to observe the morphology of myelin in the corpus callosum of rats;transmission electron microscopy was used to observe the myelin ultra-structure of rats.Results Compared with the sham-operation group,the model group and the moxibustion+LY294002 group had significant increases in Longa score(P<0.05)and escape latency(P<0.05)and significant reductions in the expression levels of the proteins associated with the PI3K/AKT/mTOR signaling pathway(P<0.05);the myelin in the corpus callosum showed unclear texture,disordered arrangement,vacuolar changes at the edge,and fragmentation with partial protrusions and disintegration,as well as a significant reduction in the number of myelinated nerve axons(P<0.05).Compared with the model group and the moxibustion+LY294002 group,the moxibustion group had significant reductions in Longa score(P<0.05)and escape latency(P<0.05)and significant increases in the expression levels of the proteins associated with the PI3K/AKT/mTOR signaling pathway(P<0.05),with certain recovery of the structure of myelin in the corpus callosum,ordered arrange-ment,dense structure at the edge,and a significant increase in the number of myelinated nerve axons(P<0.05).Conclusion Stasis-resolving and collateral-dredging moxibustion can repair myelin damage,promote myelin remodeling,and re-store white matter function by activating the PI3K/AKT/mTOR signaling pathway.

梁嘉琪;樊吟秋;石海平;乔晓迪;邓倩;郑紧紧;张庆萍

安徽中医药大学研究生院,安徽 合肥 230012安徽中医药大学第一附属医院,安徽 合肥 230012安徽中医药大学针灸推拿学院,安徽 合肥 230012

中医学

血管性痴呆;化瘀通络灸;PI3K/AKT/mTOR信号通路;髓鞘再生

Vascular dementia;Stasis-resolving and collateral-dredging moxibustion;Phosphoinositide 3 kinase/protein kinase B/mammalian target of rapamycin signaling pathway;Myelin regeneration

《安徽中医药大学学报》 2024 (002)

化瘀通络灸促VD大鼠髓鞘再生及轴突电信号传递调控机制

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国家自然科学基金项目(82104994);安徽省高校自然科学研究项目(KJ2021A0559);安徽中医药大学科研基金项目(2021yfylc15)

10.3969/j.issn.2095-7246.2024.02.015

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