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| 灯盏细辛对小鼠骨骼肌缺血再灌注损伤的作用及机制 |
| Effect and mechanism of erigeron breviscapus (Vant.) Hand-Mazz on skeletal muscle ischemia-reperfusion injury in mice |
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| DOI：10.3969/j.issn.1006-7108.2026.08.008 |
| 中文关键词: 灯盏细辛 miR-135a-5p 缺氧诱导因子1-α抑制剂 骨骼肌缺血再灌注 凋亡 |
| 英文关键词:erigeron breviscapus (Vant.) Hand-Mazz MiR-135a-5p hypoxia inducible factor 1-
alpha subunit inhibitor skeletal muscle ischemia-reperfusion apoptosis |
| 基金项目:2021年度黑龙江省省属高等学校基本科研项目（2021-KYYWF-0511） |
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| 中文摘要: |
| 目的 探讨灯盏细辛[Erigeron breviscapus (Vant.) Hand-Mazz，EBHM]对骨骼肌缺
血再灌注损伤（skeletal muscle ischemia-reperfusion injury，SMIRI）小鼠的影响及机
制。方法 将84只SPF级C57BL6雄性小鼠按照随机数字表法分为7组（12只/组）：假手
术组、Model组、EBHM低、中、高剂量组、EBHM高剂量+antagomir NC组、EBHM高剂量+miR-
135a-5p antagomir组。除假手术组外，其余6组均构建SMIRI大鼠模型。各给药组在建模
前通过灌胃、尾静脉注射相应药物，每天1次，连续7 d。假手术组组仅接受麻醉切开及伤
口缝合操作，未实施右侧股动脉环形结扎术。再灌注24 h后处死小鼠，qRT-PCR法检测miR-
135a-5p的表达；HE及TUNEL染色分别观察腓肠肌组织病理变化和细胞凋亡情况；ELISA检
测小鼠血清中炎症因子及腓肠肌组织中骨骼肌损伤因子水平；Western blot法检测Cleaved
Caspase-3及Hif1an蛋白表达；双荧光素酶报告实验验证miR-135a-5p和Hif1an之间的靶
向关系。结果 相较于假手术组，Model组miR-135a-5p表达降低，腓肠肌组织学损伤评分、
细胞凋亡率、炎症因子IL-6、IL-1β、TNF-α水平、CK-MM、LDH、MPO水平、Cleaved Caspase-
3、Hif1an蛋白表达升高（P＜0.05）；相较于Model组，EBHM低、中、高剂量组miR-135a-
5p表达依次升高，腓肠肌组织损伤学评分、细胞凋亡率、炎症因子IL-6、IL-1β、TNF-α、
CK-MM、LDH、MPO水平、Cleaved Caspase-3、Hif1an蛋白表达依次降低（P＜0.05），呈剂
量依赖性（P＜0.05）；相较于EBHM高剂量+antagomir NC组，EBHM高剂量+miR-135a-5p
antagomir组上述指标趋势均发生逆转（P＜0.05）；miR-135a-5p直接靶向Hif1an，并负
调控Hif1an蛋白表达。结论 SMIRI使小鼠腓肠肌发生细胞凋亡和炎症损伤，EBHM处理可降
低炎症因子水平和腓肠肌组织细胞凋亡，从而缓解SMIRI损伤，其机制可能与靶向上调miR-
135a-5p/Hif1an通路相关。 |
| 英文摘要: |
| Objective To investigate the effect and mechanism of erigeron breviscapus (Vant.) Hand-
Mazz (EBHM) on skeletal muscle ischemia-reperfusion injury (SMIRI) in mice. Methods Eighty-
four SPF grade C57BL6 male mice were randomly divided into 7 groups (12 mice/group) according
to the random number table method, including sham operation group, model group, EBHM low,
medium, and high dose groups, high-dose EBHM+antigomir NC group, and high-dose
EBHM+miR-135a-5p antigomir group. Except for the sham operation group, SMIRI mouse models
were constructed in the other six groups. Mice in each administration group received the
corresponding drugs by gavage and tail vein injection once a day for 7 consecutive days before
modeling. Mice in the sham operation group received anesthesia incision and wound suture
operations only, and no circular ligation of the right femoral artery was performed. The mice were
sacrificed 24 h after reperfusion. qRT-PCR method was applied to detect the expression of miR-
135a-5p. HE and TUNEL staining were applied to observe the pathological changes and cell
apoptosis of gastrocnemius muscle tissue, respectively. ELISA was applied to detect the levels of
inflammatory factors in serum and skeletal muscle injury factors in gastrocnemius tissue. Western
blotting was applied to detect the expressions of Cleaved Caspase-3 and Hif1an proteins. Dual
luciferase reporter experiment was applied to verify the targeting relationship between miR-135a-
5p and Hif1an. Results Compared to those in the sham operation group, the expression of miR-
135a-5p in the model group reduced, the histological injury score of gastrocnemius muscle, cell
apoptosis rate, levels of IL-6, IL-1β, TNF-α, CK-MM, LDH, and MPO, and the expressions of
Cleaved Caspase-3 and Hif1an proteins increased (P<0.05). Compared to those in the model group,
the expression of miR-135a-5p in the low, medium, and high dose EBHM groups increased
sequentially, the histological injury score of gastrocnemius muscle, cell apoptosis rate, levels of IL-
6, IL-1β, TNF-α, CK-MM, LDH, and MPO, and the expressions of Cleaved Caspase-3 and Hif1an
proteins decreased sequentially (P<0.05) in a dose-dependent manner (P<0.05). Compared to the
high-dose EBHM + antagomir NC group, the high-dose EBHM + miR-135a-5p antagomir group
showed a reversal trend in the aforementioned indicators. MiR-135a-5p directly targeted Hif1an and
negatively regulated the expression of Hif1an protein. Conclusion SMIRI causes apoptosis and
inflammatory injury in the gastrocnemius muscle of mice. EBHM treatment reduces the levels of
inflammatory factors and apoptosis of gastrocnemius muscle cells, thereby alleviating SMIRI injury.
The mechanism may be related to the targeted upregulation of the miR-135a-5p/Hif1an pathway. |
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