有氧运动通过调控miR-221-3p介导的脂肪组织巨噬细胞极化改善小鼠胰岛素抵抗

doi:10.12122/j.issn.1673-4254.2026.01.08

摘要/Abstract

摘要：

目的探讨微RNA 221-3p（miR-221-3p）在有氧运动改善高脂饮食诱导的脂肪组织巨噬细胞极化及胰岛素抵抗（IR）中的作用及潜在生理机制。方法 60只5周龄SPF级C57BL/6J雄性小鼠随机分为普通膳食组（20 只）和高脂膳食组（40 只）。在高脂膳食干预12周后，筛选出IR小鼠。经过造模和筛选，最终选取16只普通小鼠和16只IR小鼠进入后续实验。将小鼠随机分为4组（n=8）：正常安静组（CS组），正常运动组（CE组），高脂安静组（HS组）和高脂运动组（HE组）。运动组进行为期8周的跑台运动（坡度0°，跑速15~20 m/min，1 h/d，5 d/周）。同时对小鼠单核巨噬细胞进行miR-221-3p转染，进一步验证miR-221-3p对巨噬细胞极化的作用。实验干预结束后，检测小鼠体质量、体成分；ELISA和生化试剂盒检测小鼠空腹血糖、胰岛素及血脂4项水平；采用荧光定量PCR（qPCR）检测miR-221-3p、Socs1、Tnf-α和Arg-1相对含量；双荧光素酶报告基因实验（Luciferase）验证miR-221-3p与细胞因子信号抑制蛋白1（SOCS1）的靶向关系Western blotting检测脂肪组织SOCS1、Janus激酶1（JAK1）、磷酸化信号转导子和转录激活子1和3（p-STAT1、p-STAT3）的蛋白表达水平。结果有氧运动显著降低小鼠体质量、脂肪质量和体脂率，空腹血糖、胰岛素水平胰岛素抵抗指数（HOMA-IR）、总胆固醇（TC）、甘油三酯（TG）水平显著降低（P<0.01），瘦体质量显著升高（P<0.01）。qPCR结果显示，与CS组相比，HS组小鼠血浆和脂肪组织miR-221-3p的相对表达量均显著升高（P<0.01）；有氧运动显著降低小鼠血浆和脂肪组织miR-221-3p的相对表达量（P<0.01）。Western blotting结果显示，有氧运动显著降低了iNOS、JAK1和p-STAT1/STAT1的蛋白表达（P<0.01），Arg-1、SOCS1和p-STAT3/STAT3的蛋白表达显著提高（P<0.05，P<0.05，P<0.01）。体外实验结果显示过表达miR-221-3p可以显著降低Socs1、Arg-1的基因表达（P<0.01），miR-221-3p抑制剂显著促进了M2型巨噬细胞极化状态。结论有氧运动可能通过抑制miR-221-3p的表达，靶向激活SOCS1及下游JAK/STAT信号通路，抑制脂肪组织巨噬细胞M1极化，促进巨噬细胞M2极化，减轻脂肪组织炎症反应，从而提高组织胰岛素敏感性。

关键词: 有氧运动, 胰岛素抵抗, 微小RNA, 巨噬细胞极化

Abstract:

Objective To explore the role of miR-221-3p in mediating the positive effects of aerobic exercise on macrophage polarization in the adipose tissues and insulin resistance (IR). Methods Sixteen normal C57BL/6J mice and 16 mice with IR induced by high-fat diet (HFD) feeding for 12 weeks were both randomized into sedentary group and exercise group with aerobic exercise training on a treadmill (5 times per week for 8 consecutive weeks). All the mice were examined for changes in body weight, body composition, fasting blood glucose, blood lipid levels, insulin levels, miR-221-3p expression level, mRNA levels of Socs1, Tnf-α and Arg-1, and protein levels of SOCS1, JAK1, p-STAT1, and p-STAT3 in the adipose tissues, and the targeting relationship between miR-221-3p and SOCS1 was validated using dual-luciferase reporter gene assay. In RAW264.7 macrophages, the effects of transfection with miR-221-3p mimic or inhibitor on macrophage polarization were observed. Results In mice with normal feeding, aerobic exercise significantly decreased body weight, fat mass, fat percent, fasting blood glucose, serum insulin level, HOMA-IR, and TC and TG levels, and reduced miR-221-3p levels in both the plasma and the adipose tissues. The sedentary IR mice showed significantly increased miR-221-3p levels in both the plasma and adipose tissue, increased protein levels of iNOS, JAK1, and p-STAT1/STAT1, and decreased protein levels of Arg-1, SOCS1 and p-STAT3/STAT3, which were significantly reversed after aerobic exercise intervention. Dual-luciferase reporter gene assays validated the targeting relationship between miR-221-3p and SOCS1. In RAW264.7 macrophages, miR-221-3p overexpression significantly reduced Socs1 and Arg-1 mRNA expression, whereas miR-221-3p inhibition obviously promoted M2 polarization of the macrophages. Conclusion Aerobic exercise improves HFD-induced IR in mice possibly by inhibiting miR-221-3p to activate the SOCS1 and JAK/STAT signaling pathway, thereby promoting macrophage M2 polarization and alleviating chronic inflammation in the adipose tissue.

Key words: aerobic exercise, insulin resistance, microRNA, macrophage polarization

李楠, 张亮, 郭俏凤, 周越, 刘长江. 有氧运动通过调控miR-221-3p介导的脂肪组织巨噬细胞极化改善小鼠胰岛素抵抗[J]. 南方医科大学学报, 2026, 46(1): 74-82.

Nan LI, Liang ZHANG, Qiaofeng GUO, Yue ZHOU, Changjiang LIU. Aerobic exercise regulates macrophage polarization and improves insulin resistance in mice: the mediating role of miR-221-3p[J]. Journal of Southern Medical University, 2026, 46(1): 74-82.

图/表 9

表1 qRT-PCR引物序列

Tab.1 Primers sequences for RT-qPCR

Gene	Primer sequences (5'-3')
miR-221-3p	RT	CTCAACTGGTGTCGTGGAGTCGGCAATTCAGTTGAGGAAACCCA
miR-221-3p	F	ACACTCCAGCTGGGAGCTACATTGTCTGC
U6	F	CTCGCTTCGGCAGCACA
U6	R	AACGCTTCACGAATTTGCGT
Tnf-α	F	CCCTCACACTCAGATCATCTTCT
Tnf-α	R	GCTACGACGTGGGCTACAG
Arg-1	F	ACAGCAGAGGAGGTGAAGAGTAC
Arg-1	R	AGTCAGTCCCTGGCTTATGGT
Socs1	F	CTGCGGCTTCTATTGGGGAC
Socs1	R	AAAAGGCAGTCGAAGGTCTCG
β-actin	F	GTGCTATGTTGCTCTAGACTTCG
β-actin	R	ATGCCACAGGATTCCATACC

图1 有氧运动干预后各组小鼠体质量变化

Fig.1 Changes in body weight of the mice in each group (n=6). A: Changes in body weight. B: Changes in fat mass. C: Changes in lean body mass. D: Changes in fat percent. CS: Normally fed sedentary group without exercise; CE: Normally fed sedentary group with aerobic exercise; HS: High-fat diet sedentary group; HE: High-fat diet group with aerobic exercise. **P<0.01 vs CS group, ##P<0.01 vs HS group.

表2 有氧运动干预后各组小鼠基础指标的变化

Tab.2 Changes in the indicators of the mice in each group

Parameter	CS	CE	HS	HE
Fasting blood glucose (mmol/L)	6.70±0.38	6.13±0.44	13.10±0.99**	7.56±0.69^##
Insulin (μIU/mL)	7.74±0.62	8.27±2.70	24.73±2.41**	11.70±3.30**^##
HOMA-IR	2.32±0.22	2.24±0.72	14.46±2.27**	3.87±0.84*^##
TC (mmol/L)	3.08±0.13	3.02±0.53	4.67±0.40**	3.90±0.70**^##
TG (mmol/L)	0.59±0.07	0.60±0.09	0.88±0.19**	0.72±0.07^##
HDL (mmol/L)	2.87±0.47	2.86±0.27	3.76±0.73**	3.56±0.23
LDL (mmol/L)	0.15±0.08	0.14±0.06	0.32±0.13	0.31±0.13

图2 有氧运动干预后小鼠血液与脂肪组织中miR-221-3p变化

Fig.2 Changes in miR-221-3p levels in each group. A: Expression levels of miR-221-3p in plasma (n=6). B: Expression levels of miR-221-3p in adipose tissue (AT). **P<0.01 vs CS group, ##P<0.01 vs HS group.

图3 miR-221-3p与Socs1的靶向关系

Fig.3 Targeting relationship between Socs1 and miR-221-3p (A) and dual-luciferase activity expression (B). n=3, **P<0.01 vs WT group.

图4 细胞转染后各组miR-221-3p和Socs1的mRNA表达情况

Fig.4 Relative mRNA levels of miR-221-3p (A) and Socs1 (B) enriched in RAW 264.7 cells (n=3). **P<0.01 vs NC group, ##P<0.01 vs mimic NC group, $$P<0.01 vs inhibitor NC group.

图5 细胞转染后各组炎症因子的mRNA表达

Fig.5 Relative mRNA levels of Tnf-α and Arg-1 enriched in RAW 264.7 cells (n=3). *P<0.05, **P<0.01 vs NC group, #P<0.05, ##P<0.01 vs mimic NC group, $$P<0.01 vs inhibitor NC group.

图6 有氧运动干预后小鼠脂肪组织巨噬细胞极化相关蛋白变化

Fig.6 Changes of macrophage polarization-related proteins in the adipose tissue of mice after aerobic exercise intervention. A: Expression of iNOS proteins in adipose tissue. B: Expression of Arg-1 proteins in adipose tissue. n=6. *P<0.05, **P<0.01 vs CS group, #P<0.05, ##P<0.01 vs HS group.

图7 各组小鼠脂肪组织JAK/STAT通路相关蛋白的表达

Fig.7 Comparison of the expression levels of JAK/STAT pathway proteins SOCS1 (A), JAK1 (B), p-STAT1/STAT1 (C), and p-STAT3/STAT3 (D) in the adipose tissue of the mice. n=6. **P<0.01 vs CS group, #P<0.05, ##P<0.01 vs HS group.

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