艾灸督脉经穴通过p38 MAPK通路抑制血管性痴呆大鼠海马CA1区IL-1β和TNF-α表达

Moxibustion at Acupoints on the Governor Vessel Inhibits the Expression of Interleukin-1β and Tumor Necrosis Factor-α in the Hippocampal CA1 Region of Rats with Vascular Dementia via the p38 MAPK Pathway

目的 基于p38 MAPK通路探究艾灸督脉“百会”“大椎”“风府”穴治疗血管性痴呆的作用机制。方法 将60只SPF级雄性SD大鼠随机分为假手术组、模型组、艾灸组、阻滞剂组、艾灸+阻滞剂组。采用双侧颈总动脉永久结扎法复制血管性痴呆大鼠模型。采用Morris水迷宫实验检测大鼠认知功能,透射电子显微镜观察海马神经细胞超微结构,Western blot法和RT-qPCR法分别检测海马组织中p38丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)、肿瘤坏死因子-α(tumor necrosis factor-alpha,TNF-α)、白细胞介素-1β(interleukin-1 beta,IL-1β)蛋白及其mRNA表达水平。结果 与假手术组比较,模型组大鼠平均逃避潜伏期显著延长(P<0.05),穿越平台次数显著减少(P<0.05);海马CA1区神经细胞核染色体凝集、边集,胞浆内细胞器数量明显减少,胞浆基质完全溶解空泡化,神经纤维变形、溶解、坏死;p38 MAPK、TNF-α、IL-1β蛋白及mRNA表达水平均显著升高(P<0.05)。与模型组比较,艾灸组、阻滞剂组平均逃避潜伏期显著缩短(P<0.05),穿越平台次数显著增加(P<0.05);海马CA1区神经细胞结构有不同程度改善;p38 MAPK、TNF-α、IL-1β蛋白及mRNA表达水平均显著降低(P<0.05)。艾灸+阻滞剂组与阻滞剂组上述指标差异无统计学意义(P>0.05)。结论 艾灸对血管性痴呆大鼠的认知功能障碍具有一定的改善作用,其机制可能是通过调控p38 MAPK信号通路减少TNF-α、IL-1β蛋白表达。

Objective To investigate the mechanism of action of moxibustion at Baihui, Dazhui, and Fengfu points of the governor vessel in the treatment of vascular dementia via the p38 MAPK pathway. Methods A total of 60 specific pathogen-free male Sprague-Dawley rats were randomly divided into sham-operation group, model group, moxibustion group, blocker group, and moxibustion+blocker group. Permanent ligation of both common carotid arteries was performed to establish an animal model of vascular dementia. The Morris water maze test was used to measure cognitive function;transmission electron microscopy was used to observe the ultrastructure of hippocampal neural cells;Western blot and RT-qPCR were used to measure the protein and mRNA expression levels of mitogen-activated protein kinase (MAPK), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) in hippocampal tissue. Results Compared with the sham-operation group, the model group had a significant increase in mean escape latency ( P<0.05 ) and a significant reduction in the number of platform crossings ( P<0.05 ), with chromosome condensation and margination in the nucleus of hippocampal CA1 neurons, a significant reduction in the number of organelles in cytoplasm, complete dissolution and vacuolization of cytoplasmic matrix, and deformity, dissolution, and necrosis of nerve fibers, as well as significant increases in the protein and mRNA expression levels of p38 MAPK, TNF-α, and IL-1β ( P<0.05 ). Compared with the model group, the moxibustion group and the blocker group had a significant reduction in mean escape latency ( P<0.05 ), a significant increase in the number of platform crossings ( P<0.05 ), varying degrees of improvement in the structure of hippocampal CA1 neurons, and significant reductions in the protein and mRNA expression levels of p38 MAPK, TNF-α, and IL-1β ( P<0.05 ). There were no significant differences in the above indicators between the moxibustion+blocker group and the blocker group ( P> 0.05 ). Conclusion Moxibustion has a certain effect in improving cognitive impairment in rats with vascular dementia, possibly by regulating the p38 MAPK signaling pathway and reducing the protein expression levels of TNF-α and IL-1β.