Mechanism of Qigu capsule (芪骨胶囊) as a treatment for sarcopenia based on network pharmacology and experimental validation

doi:10.19852/j.cnki.jtcm.2025.02.001

Abstract

Abstract:

OBJECTIVE: To explore the potential molecular mechanism of Qigu capsule (芪骨胶囊，QGC) in the treatment of sarcopenia through network pharmacology and to verify it experimentally.

METHODS: The active compounds of QGC and common targets between QGC and sarcopenia were screened from databases. Then the herbs-compounds-targets network, and protein-protein interaction (PPI) network was constructed. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed by R software. Next, we used a dexamethasone-induced sarcopenia mouse model to evaluate the anti-sarcopenic mechanism of QGC.

RESULTS: A total of 57 common targets of QGC and sarcopenia were obtained. Based on the enrichment analysis of GO and KEGG, we took the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway as a key target to explore the mechanism of QGC on sarcopenia. Animal experiments showed that QGC could increase muscle strength and inhibit muscle fiber atrophy. In the model group, the expression of muscle ring finger-1 and Atrogin-1 were increased, while myosin heavy chain was decreased, QGC treatment reversed these changes. Moreover, compared with the model group, the expressions of p-PI3K, p-Akt, p-mammalian target of rapamycin and p-Forkhead box O3 in the QGC group were all upregulated.

CONCLUSION: QGC exerts an anti-sarcopenic effect by activating PI3K/Akt signaling pathway to regulate skeletal muscle protein metabolism.

Key words: sarcopenia, Network pharmacology, experimental validation, phosphatidylinositol 3-kinase, proto-oncogene proteins c-akt, signal transduction, Qigu capsule

SHI Jinyu, PAN Fuwei, GE Haiya, YANG Zongrui, ZHAN Hongsheng. Mechanism of Qigu capsule (芪骨胶囊) as a treatment for sarcopenia based on network pharmacology and experimental validation[J]. Journal of Traditional Chinese Medicine, 2025, 45(2): 399-407.

Figures/Tables 7

Table 1 Active compounds and target genes of QGC

Herb	Active compound	Target
Gusuibu (Rhizoma Drynariae)	15	122
Heshouwu (Radix Polygoni Multiflori)	16	202
Huangqi (Radix Astragali Mongolici)	25	424
Juhua (Flos Chrysanthemi)	29	626
Roucongrong (Herba Cistanches Deserticolae)	18	860
Shihu (Herba Dendrobii Nobilis)	8	60
Yinyanghuo (Herba Epimedii Brevicornus)	16	134

Table 2 Key targets of QGC in the treatment of sarcopenia (the top 10 nodes)

No.	Target	BC	CC	DC
1	AKT1	0.120718453	0.828125	43
2	INS	0.059676907	0.791045	40
3	TNF	0.031125367	0.768116	38
4	ALB	0.040891396	0.757143	37
5	IL6	0.027191572	0.757143	37
6	LEP	0.040162888	0.757143	37
7	TP53	0.120534503	0.757143	37
8	IGF1	0.023113745	0.736111	35
9	FOS	0.019708676	0.670886	30
10	MTOR	0.01690301	0.679487	29

Figure 1 Results of GO enrichment analysis and KEGG signaling pathway enrichment analysis A: results of GO enrichment analysis; B: results of KEGG signaling pathway enrichment analysis. GO: gene ontology; KEGG: kyoto encyclopedia of genes and genomes.

Table 3 Effect of QGC on body weight and grip strength ($\bar{x}±s$)

Group	n	Terminal body weight (g)	Grip strength (g)	Grip strength/body weight
NC DEX QGC	12 12 12	29.91±1.13 27.22±1.16^a 28.06±1.23	77.78±7.34 59.33±5.34^a 71.17±5.16^b	2.62±0.30 2.13±0.20^a 2.50±0.27^b

Figure 2 Effect of QGC on DEX-induced skeletal muscle atrophy A: the representative samples of dissected gastrocnemius. A1: NC group; A2: DEX group; A3: QGC group; B: ratios of gastrocnemius weight to whole body weight; C: the representative image of HE staining for gastrocnemius (× 400). C1: NC group; C2: DEX group; C3: QGC group; D: comparison of mean fiber CSA in each group; E: distribution of fiber CSA in each group. NC group: without any treatment; DEX group: treated with dexamethasone sodium phosphate 1 mg/kg body weight/day for six weeks; QGC group: treated with dexamethasone sodium phosphate 1 mg/kg body weight/day plus Qigu capsule for six weeks. NC: normal control; DEX: dexamethasone; QGC: Qigu capsule; CSA: cross-sectional areas. One‐way analysis of variance was used to compare more than two groups, followed by the LSD test to detect differences between groups. The data are presented as the mean ± standard deviation (n = 12). Compared with NC group, aP < 0.01; compared with DEX group, bP < 0.01.

Figure 3 Western blot analysis of the protein expression of muscle atrophy markers (MuRF-1, Atrogin-1) and MyHC in each group A: Western blot analysis of the protein expression of muscle atrophy markers (MuRF-1, Atrogin-1) in each group; B: comparison results of MuRF-1; C: comparison results of Atrogin-1; D: western blot analysis of the protein expression of MyHC in each group; E: comparison results of MyHC. NC group: without any treatment; DEX group: treated with dexamethasone sodium phosphate 1 mg/kg body weight/day for six weeks; QGC group: treated with dexamethasone sodium phosphate 1 mg/kg body weight/day plus qigu capsule for six weeks. NC: normal control; DEX: dexamethasone; QGC: qigu capsule; MuRF-1: muscle ring finger-1; MyHC: myosin heavy chain; GAPDH: glyceraldehyde-3-phosphate dehydrogenase. One‐way analysis of variance was used to compare more than two groups, followed by the least significant difference test to detect differences between groups. The data are presented as the mean ± standard deviation (n = 12). Compared with NC group, aP < 0.01; compared with DEX group, bP < 0.01.

Figure 4 Western blot analysis for p-PI3K, p-Akt, p-mTOR, p-FoxO3a in each group A: Western blot analysis of the protein expression of p-PI3K in each group; B: comparison results of p-PI3K.C: western blot analysis of the protein expression of p-Akt in each group; D: comparison results of p-Akt. E: western blot analysis of the protein expression of p-mTOR in each group; F: comparison results of p-mTOR. G: Western blot analysis of the protein expression of p-FoxO3a in each group. H: comparison results of p-FoxO3a. NC group: without any treatment; DEX group: treated with dexamethasone sodium phosphate 1 mg/kg body weight/day for six weeks; QGC group: treated with dexamethasone sodium phosphate 1 mg/kg body weight/day plus qigu capsule for six weeks. NC: normal control; DEX: dexamethasone; QGC: qigu capsule; PI3K: phosphatidylinositol 3-kinase; p-PI3K: phospho-phosphatidylinositol 3-kinase; Akt: serine/threonine kinase B; p-Akt: phospho-serine/threonine kinase B; mTOR: mammalian target of rapamycin; p-mTOR: phospho-mammalian target of rapamycin; FoxO3a: forkhead box O3; p-FoxO3a: phospho-forkhead box O3; GAPDH: glyceraldehyde-3-phosphate dehydrogenase. One‐way analysis of variance was used to compare more than two groups, followed by the LSD test to detect differences between groups. The data are presented as the mean ± standard deviation (n = 12). Compared with NC group, aP< 0.01; compared with DEX group, bP < 0.01; compared with DEX group, cP < 0.05.

References 39.

1.	Fielding RA, Vellas B, Evans WJ, et al. Sarcopenia: an undiagnosed condition in older adults. current consensus definition: prevalence, etiology, and consequences. International Working Group on Sarcopenia. J Am Med Dir Assoc 2011; 12: 249-56. DOI PMID
2.	Dodds RM, Syddall HE, Cooper R, et al. Grip strength across the life course: normative data from twelve British studies. PLoS One 2014; 9: e113637.
3.	Ferrucci L, de Cabo R, Knuth ND, et al. Of greek heroes, wiggling worms, mighty mice, and old body builders. J Gerontol A Biol Sci Med Sci 2012; 67A: 13-6.
4.	Frontera WR, Hughes VA, Fielding RA, et al. Aging of skeletal muscle: a 12-yr longitudinal study. J Appl Physiol (1985) 2000; 88: 1321-6.
5.	Doherty TJ. Invited review: aging and sarcopenia. J Appl Physiol (1985) 2003; 95: 1717-27.
6.	Han P, Kang L, Guo Q, et al. Prevalence and factors associated with sarcopenia in suburb-dwelling older chinese using the asian working group for sarcopenia definition. J Gerontol A Biol Sci Med Sci 2016; 71: 529-35.
7.	Landi F, Cruz-Jentoft AJ, Liperoti R, et al. Sarcopenia and mortality risk in frail older persons aged 80 years and older: results from ilSIRENTE study. Age Ageing 2013; 42: 203-9. DOI PMID
8.	Landi F, Russo A, Liperoti R, et al. Midarm muscle circumference, physical performance and mortality: results from the aging and longevity study in the sirente geographic area (ilSIRENTE study). Clin Nutr 2010; 29: 441-7. DOI PMID
9.	Yalcin A, Aras S, Atmis V, et al. Sarcopenia and mortality in older people living in a nursing home in Turkey. Geriatr Gerontol Int 2017; 17: 1118-24. DOI PMID
10.	Zheng Y, Wang X, Zhang Z, et al. Bushen Yijing Fang reduces fall risk in late postmenopausal women with osteopenia: a randomized double-blind and placebo-controlled trial. Sci Rep 2019; 9.2089.
11.	Hopkins AL. Network pharmacology. Nat Biotechnol 2007; 25: 1110-1. DOI PMID
12.	Wu X, Jiang R, Zhang MQ, et al. Network-based global inference of human disease genes. Mol Syst Biol 2008; 4: 189. DOI PMID
13.	Nair A, Jacob S. A simple practice guide for dose conversion between animals and human. J Basic Clin Pharm 2016; 7: 27-31.
14.	Cohen S, Nathan JA, Goldberg AL. Muscle wasting in disease: molecular mechanisms and promising therapies. Nat Rev Drug Discov 2015; 14: 58-74. DOI PMID
15.	Rolland Y, Czerwinski S, Abellan VKG, et al. Sarcopenia: its assessment, etiology, pathogenesis, consequences and future perspectives. J Nutr Health Aging 2008; 12: 433-50. DOI PMID
16.	Iellamo F, Volterrani M, Caminiti G, et al. Testosterone therapy in women with chronic heart failure: a pilot double-blind, randomized, placebo-controlled study. J Am Coll Cardiol 2010; 56: 1310-6. DOI PMID
17.	Kovacheva EL, Hikim AP, Shen R, et al. Testosterone supplementation reverses sarcopenia in aging through regulation of myostatin, c-Jun NH2-terminal kinase, Notch, and Akt signaling pathways. Endocrinology 2010; 151: 628-38. DOI PMID
18.	Egerman MA, Glass DJ. Signaling pathways controlling skeletal muscle mass. Crit Rev Biochem Mol Biol 2013; 49: 59-68.
19.	Glass DJ. Molecular mechanisms modulating muscle mass. Trends Mol Med 2003; 9: 344-50. DOI PMID
20.	Hers I, Vincent EE, Tavaré JM. Akt signalling in health and disease. Cell Signal 2011; 23: 1515-27. DOI PMID
21.	Clemmons DR. Role of IGF-I in skeletal muscle mass maintenance. Trends Endocrinol Metab 2009; 20: 349-56.
22.	Gruner S, Peter D, Weber R, et al. The structures of eIF4E-eIF4G complexes reveal an extended Interface to regulate translation Initiation. Mol Cell 2016; 64: 467-79. DOI PMID
23.	Ruegg MA, Glass DJ. Molecular mechanisms and treatment options for muscle wasting diseases. Annu Rev Pharmacol Toxicol 2011; 51: 373-95. DOI PMID
24.	Bodine SC, Latres E, Baumhueter S, et al. Identification of ubiquitin ligases required for skeletal muscle atrophy. Science 2001; 294: 1704-8. DOI PMID
25.	Gumucio JP, Mendias CL. Atrogin-1, MuRF-1, and sarcopenia. Endocrine 2013; 43: 12-21. DOI PMID
26.	Sandri M, Sandri C, Gilbert A, et al. Foxo transcription factors induce the atrophy-related ubiquitin ligase atrogin-1 and cause skeletal muscle atrophy. Cell 2004; 117: 399-412. DOI PMID
27.	Mammucari C, Milan G, Romanello V, et al. FoxO3 controls autophagy in skeletal muscle in vivo. Cell Metab 2007; 6: 458-71. DOI PMID
28.	Zhao J, Brault JJ, Schild A, et al. FoxO3 coordinately activates protein degradation by the autophagic/lysosomal and proteasomal pathways in atrophying muscle cells. Cell Metab 2007; 6: 472-83. DOI PMID
29.	Brunet A, Bonni A, Zigmond MJ, et al. Akt promotes cell survival by phosphorylating and inhibiting a forkhead transcription factor. Cell 1999; 96: 857-68. DOI PMID
30.	Stitt TN, Drujan D, Clarke BA, et al. The IGF-1/PI3K/Akt pathway prevents expression of muscle atrophy-induced ubiquitin ligases by inhibiting FOXO transcription factors. Mol Cell 2004; 14: 395-403. DOI PMID
31.	Kaasik P, Umnova M, Pehme A, et al. Ageing and dexamethasone associated sarcopenia: peculiarities of regeneration. J Steroid Biochem Mol Biol 2007; 105: 85-90.
32.	Xie WQ, He M, Yu DJ, et al. Mouse models of sarcopenia: classification and evaluation. J Cachexia Sarcopenia Muscle 2021; 12: 538-54.
33.	Kim S, Kim K, Park J, et al. Curcuma longa L. water extract improves dexamethasone-induced sarcopenia by modulating the muscle-related gene and oxidative stress in mice. Antioxidants (Basel) 2021; 10: 1000.
34.	Ma J, Ye M, Li Y, et al. Zhuanggu Zhitong capsule alleviates osteosarcopenia in rats by up-regulating PI3K/Akt/Bcl 2 signaling pathway. Biomed Pharmacother 2021; 142: 111939.
35.	Lala-Tabbert N, Lejmi-Mrad R, Timusk K, et al. Targeted ablation of the cellular inhibitor of apoptosis 1 (cIAP1) attenuates denervation-induced skeletal muscle atrophy. Skelet Muscle 2019; 9: 13. DOI PMID
36.	Kim YI, Lee H, Nirmala FS, et al. Antioxidant activity of valeriana fauriei protects against dexamethasone-induced muscle atrophy. Oxid Med Cell Longev 2022; 2022: 3645431.
37.	Clarke BA, Drujan D, Willis MS, et al. The E3 ligase MuRF1 degrades myosin heavy chain protein in dexamethasone-treated skeletal muscle. Cell Metab 2007; 6: 376-85. PMID
38.	Lin YA, Li YR, Chang YC, et al. Activation of IGF-1 pathway and suppression of atrophy related genes are involved in epimedium extract (icariin) promoted C2C 12 myotube hypertrophy. Sci Rep 2021; 11: 10790.
39.	Zhang ZK, Li J, Liu J, et al. Icaritin requires phosphatidylinositol 3 kinase (PI3K)/Akt signaling to counteract skeletal muscle atrophy following mechanical unloading. Sci Rep 2016; 6: 20300.

[1]	FENG Guiling, ZHOU Xiaolin, SHEN Chengwan, LI Panxiao, ABULIZI ·Abudula. Effects of Huluan decotion (护卵汤) on cyclophosphamide-induced autoimmune premature ovarian failure in murine models [J]. Journal of Traditional Chinese Medicine, 2025, 45(2): 266-271.
[2]	HUANG Haiyang, ZHU Shumin, ZHONG Shaowen, LIU Ying, HOU Shaozhen, GAO Jie, OU Jianzhao, DONG Mingguo, NING Weimin. Fuzheng Xuanfei Huashi prescription (扶正宣肺化湿方) suppresses inflammation in lipopolysaccharide-induced lung injury in mice via toll-like recptor 4/nuclear transcription factor κB and cyclooxygenase-2/prostaglandin E2 pathway [J]. Journal of Traditional Chinese Medicine, 2025, 45(2): 272-280.
[3]	CHEN Xi, QU Tiange, JIA Hui, DUAN Xingwu, LI Jianhong, ZHANG Kaihui, ZHANG Runtian, WANG Ruijie. Effects of Huoxue Chubi decoction (活血除痹汤) on protein kinase B-mammalian target of rapamycin autophagy pathway in scleroderma Balb/c model mice [J]. Journal of Traditional Chinese Medicine, 2025, 45(2): 303-310.
[4]	HUANG Jiaen, LUO Qing, DONG Gengting, PENG Weiwen, HE Jianhong, DAI Weibo. Xiahuo Pingwei San (夏藿平胃散) attenuated intestinal inflammation in dextran sulfate sodium-induced ulcerative colitis mice through inhibiting the receptor for advanced glycation end-products signaling pathway [J]. Journal of Traditional Chinese Medicine, 2025, 45(2): 311-325.
[5]	HAN Shuai, Du Zhikang, WANG Zirui, HUANG Tianfeng, GE Yali, SHI Jianwen, GAO Ju. Network pharmacology approach to unveiling the mechanism of berberine in the amelioration of morphine tolerance [J]. Journal of Traditional Chinese Medicine, 2025, 45(2): 376-384.
[6]	TAN Xiying, GU Ruxin, TAO Jing, ZHANG Yu, SUN RuiQian, YIN Gang, ZHANG Shuo, TANG Decai. Integrating network pharmacology and experimental validation to uncover the synergistic effects of Huangqi (Radix Astragali Mongolici)-Ezhu (Rhizoma Curcumae Phaeocaulis) with 5-fluorouracil in colorectal cancer models [J]. Journal of Traditional Chinese Medicine, 2025, 45(2): 385-398.
[7]	HU Huiming, WENG Jiajun, TANG Fangrui, WANG Yaqi, FAN Shengxian, WANG Xuecheng, CUI Can, SHAO Feng, ZHU Yanchen. Hypolipidemic effect and mechanism of Hedan tablet (荷丹片) based on network pharmacology [J]. Journal of Traditional Chinese Medicine, 2025, 45(2): 408-421.
[8]	YUAN Jianan, CHENG Kunming, LI Chao, ZHANG Xiang, DING Zeyu, LI Bing, ZHENG Yongqiu. Atractylenolide I ameliorates post-infectious irritable bowel syndrome by inhibiting the polymerase I and transcript release factor and c-Jun N-terminal kinase/inducible nitric oxide synthase pathway [J]. Journal of Traditional Chinese Medicine, 2025, 45(1): 57-65.
[9]	YAN Kai, WANG Wei, WANG Yan, GAO Huijuan, FENG Xingzhong. Network pharmacology-based study on the mechanism of Tangfukang formula (糖复康方) against type 2 diabetes mellitus [J]. Journal of Traditional Chinese Medicine, 2025, 45(1): 76-88.
[10]	ZHU Peixuan, SU Zeqi, FAN Qiongyin, ZHANG Cai, WANG Ting. Network pharmacology and animal experiments revealed the protective effects of Guilong prescription (归龙方) on chronic prostatitis and its possible mechanisms [J]. Journal of Traditional Chinese Medicine, 2025, 45(1): 89-99.
[11]	LUO Kun, ZHONG Yumei, GUO Yanding, ZHANG Linlin, HU Danhui, MA Wenbin, YANG Xin, ZHOU Haiyan. Moxibustion inhibits the macrophage M1 polarization toll-like receptor 4/myeloid differentiation factor 88/nuclear factor kappa B signaling pathway by regulating T-cell immunoglobulin and mucin-containing protein-3 in rheumatoid arthritis [J]. Journal of Traditional Chinese Medicine, 2024, 44(6): 1227-1235.
[12]	ZHANG Guangshun, XU Xiaonan, XU Chuyun, LIAO Guanghui, XU Hao, LOU Zhaohuan, ZHANG Guangji. Actinidia chinensis polysaccharide interferes with the epithelial-mesenchymal transition of gastric cancer by regulating the nuclear transcription factor-κB pathway to inhibit invasion and metastasis [J]. Journal of Traditional Chinese Medicine, 2024, 44(5): 896-905.
[13]	YANG Chunyan, LUO Jia, PENG Weijie, DAI Weibo. Huaiyu pill (槐榆片) alleviates inflammatory bowel disease in mice via blocking toll like receptor 4/ myeloid differentiation primary response gene 88/ nuclear factor kappa B subunit 1 pathway [J]. Journal of Traditional Chinese Medicine, 2024, 44(5): 916-925.
[14]	YU Siyun, ZHANG Shiwen, XIA Yu, LIU Xiaoqing, LIU Yajie, FU Jinrong. Network-based pharmacology and experimental validation to explore the mechanism of action of the Jiawei Pentongling formula (加味盆痛灵方) for the treatment of endometriosis-related pain [J]. Journal of Traditional Chinese Medicine, 2024, 44(5): 991-999.
[15]	GUO Yanding, LUO Kun, ZHANG Linlin, LU Wenting, SHANG Yanan, ZHONG Yumei, HU Danhui, YANG Xin, ZHOU Haiyan. Study on the anti-inflammatory mechanism of moxibustion in rheumatoid arthritis in rats based on phospholipaseA2 signaling inhibition by Annexin 1 [J]. Journal of Traditional Chinese Medicine, 2024, 44(4): 753-761.

Home

Online First

Author Center

Reviewer Center

Issues

Announcement

About Us