Exploring the mechanism of action of Cistanche deserticola in treating liver fibrosis based on network pharmacology, molecular docking, and experimental verification
The liver, as an important digestive gland in the human body, once diseased, will affect the normal metabolic function of the body. Research shows that the important pathway for the formation of liver cancer is liver fibrosis. When the liver is stimulated, immune cells secrete a large amount of immune regulatory factors, which activate and proliferate hepatic stellate cells, synthesize a large amount of collagen, deposit a large amount of extracellular matrix in the liver, and cause fibrosis formation. Therefore, controlling at any stage of liver disease progression may alleviate the damage of liver disease, and research on treating liver fibrosis is of great importance. In recent years, studies have shown that Cistanche deserticola has significant therapeutic effects on liver disease and shows obvious advantages in the treatment of liver fibrosis.
Cistanches Herba, an authentic medicinal herb in Xinjiang, is known as the “desert ginseng” and has been clinically used for thousands of years. Modern research has shown that it has various biological activities such as anti-aging, protecting the liver, relieving fatigue, anti osteoporosis, moisturizing the intestines and promoting bowel movements. Studies have shown that phenylethanolic acid glycosides from Cistanche deserticola can alleviate the degree of liver fibrosis. Zhang et al. found that the activation of hepatic stellate cells and liver fibrosis can be inhibited by Cistanche deserticola phenylethanolic glycoside liposomes. However, the pharmacological effects and mechanisms of Cistanche deserticola in treating liver fibrosis are currently unclear and need to be elucidated.
This study applies network pharmacology to construct a “component disease target pathway” network diagram, and then conducts protein interaction and target enrichment analysis to ultimately predict the relevant targets and pathways for the treatment of liver fibrosis with Cistanche deserticola. Through molecular docking technology and animal experiments, it provides a basis for the pharmacological effects of effective components of Cistanche deserticola in the treatment of liver fibrosis.

The development process of liver fibrosis is complex, and various cytokines and cellular pathways interact and influence each other, forming a complex network of interactions. Therefore, single target and single pathway therapy is difficult to take effect. Traditional Chinese medicine has the characteristics of multiple components, multiple targets of action, and complex pathways of action, which systematically regulate the human body. Cistanche deserticola has a protective effect on the liver. The effective ingredients of Cistanche deserticola were obtained from databases such as the Traditional Chinese Medicine Database (TCMSP), including verbascoside, echinacoside, β – sitosterol, arachidonic acid, suzilactone, goosefoot resin phenol B dimethyl ether, quercetin, and dipyridamole. There are studies indicating that phenylethanoid glycosides are the main active ingredients in Cistanche deserticola, among which echinacoside and verbascoside are considered the main bioactive components that exert pharmacological effects in Cistanche deserticola. Through network pharmacology, we obtained targets related to liver fibrosis that act on verbascoside and verbascoside, and validated them through molecular docking.
Pinecone chrysanthemum glycoside and verbascoside, as the main components of Cistanche deserticola, exert anti liver fibrosis effects by regulating apoptosis of hepatic stellate cells. ICAM1, AKT, and MMP9 may be targets of action for the treatment of liver fibrosis with echinacoside and verbascoside. As a cell adhesion factor, ICAM1 is highly expressed in cases of liver injury and can be inhibited by suppressing ICAM1 overexpression to prevent the development of liver fibrosis. Serine/threonine protein kinase (AKT) is an important target kinase downstream of PI3K. Activated AKT further activates and promotes the proliferation of hepatic stellate cells, accelerating the progression of liver fibrosis. Conversely, inhibiting AKT inhibits the occurrence of liver fibrosis; MMP-9 can degrade the normal liver collagen matrix, disrupt the intracellular environment of liver cells, promote the activation of hepatic stellate cells, secrete a large amount of collagen, and promote the development of liver fibrosis. Conversely, inhibiting MMP-9 can inhibit the occurrence of liver fibrosis; The use of CCl4 to establish a mouse model of liver fibrosis in this experiment is one of the classic modeling methods. CCl4 is injected intraperitoneally into the animal body, and compared with the normal group, the LN, HA, PCIII, and 13Col IV contents in the model group mice are significantly increased; Compared to the model group, the high, medium, and low dose groups of CPhGs significantly reduced the levels of LN, HA, PCIII, and Col IV; The results of HE and Masson tests showed that compared with the model group, the high, medium, and low dose groups of CPhGs had milder liver cell necrosis, significantly reduced eosinophil granules, and significantly reduced blue fibers around the portal area. The above results indicate that the inhibition of liver fibrosis progression by total glycosides of phenylethanol in Cistanche deserticola may be related to ICAM1, AKT, and MMP9 targets, and further detection of related protein expression is needed.
The generation of liver fibrosis is caused by many factors, among which PI3K-AKT pathway is one of the most important ones obtained by pathway enrichment analysis. PI3K-AKT signaling pathway is involved in regulating cell proliferation and apoptosis, inducing human tumorigenesis, transcription of hepatitis B virus genes, and is one of the important pathways for treating liver fibrosis. The PI3K-AKT pathway can activate the formation of hepatic blood vessels, improve blood supply, rebuild extracellular matrix, regulate autophagy, inhibit hepatocyte apoptosis, promote hepatocyte proliferation, and ultimately lead to liver fibrosis. Pinecone chrysanthemum glycoside and verbascoside can inhibit the proliferation and activation of liver cells by regulating ICAM1, AKT, and MMP9 targets, respectively. Cistanche deserticola phenylethanol total glycoside may intervene in the progression of liver fibrosis by participating in energy metabolism, proliferation, and apoptosis of liver cells. Real time fluorescence quantitative PCR detection showed that compared with the normal group, the expression levels of PI3K and AKT mRNA in the liver tissue of the model group mice were significantly upregulated; Compared with the model group, the expression levels of PI3K and AKT mRNA were significantly reduced in the high, medium, and low dose groups of total phenylethanol glycosides in Cistanche deserticola, which may inhibit the activation of the PI3K-AKT pathway and delay the progression of liver fibrosis.
In summary, the total glycosides of phenylethanol in Cistanche deserticola can effectively alleviate CCl4 induced liver fibrosis in mice, and its mechanism may be related to the proliferation and activation of hepatic stellate cells mediated by the PI3K-AKT signaling pathway. The results of this study lay the foundation for further exploration of the pharmacological effects of Cistanche deserticola in traditional Chinese medicine.