1.湖北中医药大学 针灸骨伤学院,武汉 430061
2.湖北省中医院/湖北中医药大学 附属医院,武汉 430061
3.中医肝肾研究及应用湖北省重点实验室(湖北省中医院),武汉 430061
4.湖北时珍实验室,武汉 430061
石龙,在读硕士,从事中医药防治肌肉骨骼疾病研究,E-mail:Monooxx@hotmail.com
张志文,主任医师,博士,博士生导师,从事中西医结合防治肌肉骨骼疾病研究,E-mail:zzwjjdd@163.com
收稿:2025-05-28,
修回:2025-08-13,
录用:2025-08-13,
网络出版:2025-08-22,
纸质出版:2026-02-05
移动端阅览
石龙,李扬,颜鸿宇等.基于SIRT1/Nrf2/HO-1信号通路探讨二精丸改善肌少症骨骼肌细胞炎症反应和细胞凋亡的机制[J].中国实验方剂学杂志,2026,32(03):57-66.
SHI Long,LI Yang,YAN Hongyu,et al.Erjingwan Alleviate Inflammatory Response and Apoptosis in Skeletal Muscle Cells of Sarcopenia via SIRT1/Nrf2/HO-1 Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2026,32(03):57-66.
石龙,李扬,颜鸿宇等.基于SIRT1/Nrf2/HO-1信号通路探讨二精丸改善肌少症骨骼肌细胞炎症反应和细胞凋亡的机制[J].中国实验方剂学杂志,2026,32(03):57-66. DOI: 10.13422/j.cnki.syfjx.20251704.
SHI Long,LI Yang,YAN Hongyu,et al.Erjingwan Alleviate Inflammatory Response and Apoptosis in Skeletal Muscle Cells of Sarcopenia via SIRT1/Nrf2/HO-1 Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2026,32(03):57-66. DOI: 10.13422/j.cnki.syfjx.20251704.
目的
2
探讨经典中药复方二精丸通过调控沉寂信息调节因子1(SIRT1)/核因子E
2
相关因子2(Nrf2)/血红素加氧酶-1(HO-1)信号通路对肌少症小鼠模型骨骼肌细胞炎症反应和细胞凋亡的影响及其机制。
方法
2
将40只C57/BL6雄性小鼠随机分为正常组,模型组,二精丸低、中、高剂量组(8、16、32 g·kg
-1
),采用
D
-半乳糖诱导骨骼肌衰老建立肌少症模型。检测不同剂量二精丸影响下小鼠体质量、握力以评估其生理功能;苏木素-伊红(HE)染色及马松(Masson)染色观察小鼠骨骼肌病理变化及纤维化程度;酶联免疫吸附测定法(ELISA)检测小鼠血清中肿瘤坏死因子-
α
(TNF-
α
)、白细胞介素-6(IL-6)含量,生化检测小鼠血清中超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽(GSH)水平;蛋白免疫印迹法(Western blot)及实时荧光定量聚合酶链式反应(Real-time PCR)检测各组小鼠SIRT1、Nrf2、B细胞淋巴瘤-2(Bcl-2)相关X蛋白(Bax)、Bcl-2蛋白表达量及mRNA表达水平。
结果
2
药物干预4周后,与正常组比较,模型组体质量、握力降低(
P
0.01);与模型组比较,二精丸各剂量组小鼠体质量均在第8周显著上升(
P
0.01),握力自第6周起显著上升(
P
0.01)。HE染色结果显示,正常组肌纤维结构清晰;模型组肌纤维断裂萎缩;二精丸组肌纤维结构呈剂量依赖性修复。Masson染色显示,正常组胶原纤维少、纤维化轻;模型组胶原纤维增生、纤维化重;二精丸组胶原增生且呈剂量依赖性抑制纤维化。ELISA检测结果显示,与正常组比较,模型组小鼠血清中TNF-
α
、IL-6的含量显著升高(
P
0.01);与模型组比较,二精丸低剂量组小鼠血清中TNF-
α
含量下降(
P
0.05);二精丸中、高剂量组小鼠血清中TNF-
α
、IL-6的含量均显著降低(
P
0.01)。生化检测结果显示,与正常组比较,模型组中SOD、GSH水平显著下降(
P
0.01),MDA水平显著升高(
P
0.01);与模型组比较,二精丸中、高剂量组中SOD、GSH水平显著升高(
P
0.01),MDA水平显著下降(
P
0.01)。Western blot及Real-time PCR检测结果显示,与正常组比较,模型组小鼠肌肉组织SIRT1、Nrf2、HO-1、Bcl-2蛋白及mRNA表达水平显著降低(
P
0.01),而Bax蛋白及mRNA表达水平则显著升高(
P
0.01)。与模型组比较,经二精丸干预后,各剂量组小鼠肌肉组织中SIRT1、Nrf2、HO-1、Bcl-2蛋白的表达水平均显著升高(
P
0.01);Bax蛋白及mRNA的表达水平均显著降低(
P
0.01),低剂量组Nrf2、HO-1 mRNA表达水平均升高(
P
0.05,
P
0.01),中、高剂量组SIRT1、
Nrf2、HO-1、Bcl-2 mRNA表达水平均显著升高(
P
0.01)。
结论
2
二精丸可降低肌少症小鼠模型骨骼肌细胞炎症因子含量,提高抗氧化能力,减轻肌肉病理变化和纤维化程度,其机制可能与SIRT1/Nrf2/HO-1通路协同调控炎症反应及细胞凋亡网络有关。
Objective
2
To investigate the effects of the classical Chinese medicine compound prescription Erjingwan on the inflammatory response and apoptosis of skeletal muscle cells in a mouse model of sarcopenia and decipher the mechanism based on the silent information regulator 1 (SIRT1)/nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling pathway.
Methods
2
Forty C57/BL6 male mice were randomized into a control group, a model group, and groups with different doses of Erjingwan (8,16,32 g·kg
-1
). The mouse model of sarcopenia was established by
D
-gal-induced skeletal muscle senescence. The body weight and grip strength of mice treated with different doses of Erjingwan were examined to evaluate their physiological functions. Hematoxylin-eosin (HE) staining and Masson staining were used to observe the pathological changes and fibrosis in the skeletal muscle of mice. Enzyme-linked immunosorbent assay (ELISA) was adopted to determine the levels of tumor necrosis factor-
α
(TNF-
α
) and interleukin-6 (IL-6) in the serum samples of mice, and biochemical tests were conducted to quantify the levels of superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione (GSH) in the serum. The protein and mRNA levels of SIRT1, Nrf2, B-cell lymphoma (Bcl-2), and Bcl-2-associated X protein (Bax) were determined by Western blot and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR), respectively.
Results
2
After 4 weeks of drug intervention, the model group exhibited significant reductions in body weight and grip strength (
P
0.01) compared with the control group. Compared wi
th the model group, all doses of Erjingwan increased the body weight in mice at week 8 (
P
0.01) and grip strength from week 6 (
P
0.01). HE staining revealed clear muscle fiber structure in the control group, muscle fiber rupture and atrophy in the model group, and dose-dependent repair of muscle fiber structure in the Erjingwan groups. Masson staining showed minimal collagen fibers and mild fibrosis in the control group, collagen fiber proliferation and severe fibrosis in the model group, and collagen proliferation with dose-dependent inhibition of fibrosis in the Erjingwan groups. ELISA results showed that serum levels of TNF-
α
and IL-6 were elevated in the model group compared with those in the control group (
P
0.01). After intervention, the low-dose Erjingwan group exhibited a decreased TNF-
α
level (
P
0.05), while the medium and high-dose groups showed decreases in both TNF-
α
and IL-6 levels (
P
0.01). Biochemical assays revealed that the model group had decreased SOD and GSH levels (
P
0.01) and an increased MDA level (
P
0.01) compared with the control group. The medium and high-dose Erjingwan groups exhibited increases in SOD and GSH levels (
P
0.01) and decreases in MDA level (
P
0.01), compared with the model group. WB and Real-time PCR results showed that compared with the control group, the model group presented down-regulated protein and mRNA levels of SIRT1, Nrf2, HO-1, and Bcl-2 in the muscle tissue (
P
0.01) and up-regulated protein and mRNA levels of Bax (
P
0.01). Compared with the model group, Erjingwan at different doses up-regulated the protein levels of SIRT1, Nrf2, HO-1, and Bcl-2
(
P
0.01) and down-regulated the protein and mRNA levels of Bax (
P
0.01) in the muscle tissue. Low-dose Erjingwan elevated the mRNA levels of Nrf2 and
HO-1 (
P
0.05,
P
0.01), and medium and high-dose Erjingwan up-regulated the mRNA levels of SIRT1, Nrf2, HO-1, and Bcl-2 (
P
0.01).
Conclusion
2
Erjingwan reduced the content of inflammatory factors in skeletal muscle cells, improved the antioxidant capacity, and attenuated pathological changes and fibrosis in the muscle of the mouse model of sarcopenia by regulating the SIRT1/Nrf2/HO-1 pathway, inflammatory response, and apoptosis network.
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