温和艾灸通过上调MALAT1抑制miR-125b改善缺血再灌注小鼠的脑损伤和神经细胞凋亡*

作者：贺涟漪，陈 青，王晓倩，张月娟

单位：湖南中医药大学第一附属医院，湖南 长沙 410007

引用：引用：贺涟漪，陈青，王晓倩，张月娟.温和艾灸通过上调MALAT1抑制miR-125b改善缺血再灌注小鼠的脑损伤和神经细胞凋亡[J].中医药导报,2025,31(6):67-72.

DOI：10.13862/j.cn43-1446/r.2025.06.012

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摘要：目的：探讨温和艾灸对于脑梗死的治疗效果，并基于MALAT1与miR-125b揭示其作用机制。方法：将70只小鼠随机分为Sham组、Model组、Model+温和艾灸组、Model+温和艾灸+sh-NC组、Model+温和艾灸+sh-MALAT1组、Model+温和艾灸+sh-MALAT1+inhibitor-NC组、Model+温和艾灸+sh-MALAT1+miR-125b-inhibitor组，每组10只，除Sham组外，其余各组均构建大脑动脉闭塞（MCAO）模型。采用改良神经功能评分（mNSS评分）、TUNEL染色与TTC染色判断造模是否成功；Western blotting检测cleaved Caspase-3、Bax和Bcl-2的表达；ELISA检测IL-6和TNF-α的水平；RT-qPCR检测MALAT1和miRNA-125b的表达。在细胞实验中，通过双荧光素酶报告基因实验评估MALAT1与miR-125b的结合关系。结果：建立MCAO模型后，mNss评分升高、cleaved Caspase-3、Bax和miRNA-125b表达升高（P<0.05），IL-6和TNF-α水平增加（P<0.05），而Bcl-2和MALAT1表达降低（P<0.05）。温和艾灸治疗后，mNSS评分降低，并显著降低了cleaved Caspase-3、Bax和miRNA-125b的表达以及IL-6和TNF-α的水平（P<0.05），但可上调Bcl-2和MALAT1的表达（P<0.05）。双荧光素酶报告基因实验表明MALAT1可以靶向抑制miR-125b。同时，敲低MALAT1可以显著逆转温和艾灸的治疗作用，进一步抑制miR-125b表达阻断了MALAT1敲低的不利作用。结论：温和艾灸可通过上调MALAT1靶向抑制miR-125b，从而改善缺血再灌注小鼠的脑损伤和神经细胞凋亡。

关键词：脑梗死；艾灸；温和灸；大脑动脉闭塞；MALAT1；miR-125b

Abstract：Objective: To investigate the therapeutic effect of mild moxibustion on cerebral infarction and to reveal its mechanism of action based on MALAT1 and miR-125b. Methods: Totally 70 rats were randomly divided into Sham group, Model group, Model+mild moxibustion group, Model+mild moxibustion+sh-NC group, Model+mild moxibustion+sh-MALAT1 group, Model+mild moxibustion+sh-MALAT1+inhibitor-NC group, and Model+mild moxibustion+sh-MALAT1+miR-125b-inhibitor group, 10 rats in each group. Except for the Sham group, the MCAO mouse model was constructed. Modified neurological severity score (mNSS score), TUNEL staining and TTC staining were used to determine  the success of modeling. Western blotting was used to detect the expression of cleaved Caspase-3, Bax and Bcl-2. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of IL-6 and TNF-α. RT-qPCR was used to detect the expression of MALAT1 and miRNA-125b. In cell experiments, the binding relationship between MALAT1 and miR-125b was assessed by a dual luciferase reporter gene assay. Results: Establishment of the MCAO model was associated with elevated mNSS scores,  elevated cleaved Caspase-3, Bax, and miRNA-125b expression (P<0.05), and increased levels of IL-6 and TNF-α (P<0.05), whereas Bcl-2 and MALAT1 expression were decreased (P<0.05). Mild moxibustion treatment decreased mNSS scores and reduced the expression of cleaved Caspase-3, Bax, and miRNA-125b as well as the levels of IL-6 and TNF-α (P<0.05), while it upregulated the expression of Bcl-2 and MALAT1 (P<0.05). Dual luciferase reporter gene experiments demonstrated that MALAT1 could target and inhibit miR-125b. Meanwhile, knockdown of MALAT1 significantly reversed the therapeutic effect of mild moxibustion, and further inhibition of miR-125b expression blocked the detrimental effect of MALAT1 knockdown. Conclusion: Mild moxibustion can ameliorate brain injury and neuronal apoptosis in ischaemia-reperfusion mice by up-regulating MALAT1-targeted inhibition of miR-125b.

Key words：cerebral infarction; moxibustion; mild moxibustion; MCAO; MALAT1; miR-125b

发布时间：2026-01-03

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