Analysis of the mechanism of action of Danshen Chuanxiong medicine in the treatment of Alzheimer’s disease based on network pharmacology
With the increasing aging of the world population, the incidence of Alzheimer’s disease (AD) is also increasing year by year, and it is estimated that it will increase to 115 million by 2050, posing a major threat to the health and quality of life of the elderly, and bringing enormous pressure to families and society. Due to the insidious onset, long course, complex clinical manifestations and pathogenesis of AD, the cure rate of modern Western medicine is extremely low, and it is prone to drug resistance and has many side effects. Traditional Chinese medicine has a long history of treating AD and its efficacy has been recognized.

In the Qing Dynasty, Tang Rongchuan’s “Blood Syndrome Theory” stated that “if blood is on top, it will be turbid and unclear.” Wang Qingren’s “Medical Forest Correction of Mistakes” stated, “If there is blood stasis, it will be forgotten easily. Therefore, blood stasis is an important pathogenic factor of dementia, and promoting blood circulation and removing blood stasis is one of the basic treatment methods for AD. Yan et al. found in their study of traditional Chinese medicine literature that commonly used drugs in the treatment of Alzheimer’s disease were effective in promoting blood circulation and removing blood stasis. Among the 90 basic formulas for treating Alzheimer’s disease, Dan was the most frequently used in combination with Chuanxiong, suggesting that Danshen Chuanxiong may have a good therapeutic effect on AD. This study applies network pharmacology methods to explore the mechanism of action of Danshen Chuanxiong medicine in the treatment of AD, in order to provide valuable information for the clinical treatment of AD with the method of promoting blood circulation and removing blood stasis.

AD belongs to the category of “dementia” and “dementia” in traditional Chinese medicine. Elderly people gradually lose the function of their five organs, lack the ability to promote blood circulation due to qi deficiency, and have poor blood circulation. They are prone to stasis and obstruction of the meridians, causing stagnation of the brain and body, loss of nourishment of the spirit, and dysfunction of the divine mechanism, ultimately resulting in dementia. As stated in the “Medical Zhong Zhong Shen Xi Lu”: “Brain meridian stasis can directly damage the syndrome.” As stated in the “Medical Zhong Zhong Shen Xi Lu”: “Brain meridian stasis can directly damage the brain marrow, gradually empty the brain orifices, gradually lose spiritual memory, and eventually become a dementia syndrome.” Therefore, blood stasis is an important pathogenic factor of AD, and the method of promoting blood circulation and removing blood stasis is one of the important treatment methods for preventing and treating AD. Danshen has the effects of promoting blood circulation, removing blood stasis, and calming the mind. On the day of “Dian Nan Ben Cao”: “Nourish the heart and set the mind, calm the mind and soothe the heart, treat forgetfulness, palpitations, and insomnia. Chuanxiong has the effects of promoting blood circulation, dispelling wind, and relieving pain. The “Bencao Tongxuan” records that it can “dehumidify and stop diarrhea, promote qi circulation and open up stagnation, remove blood stasis and generate new energy”. When the blood is in great motion, it cannot be generated; when the blood is not in motion, it stops and cannot be generated. “Although Chuanxiong Xin San produces blood, its beauty lies in the motion, not in the dispersion. Xing Qi opens up stagnation, enhances the clear yang qi, removes dampness from the head, is good at unblocking, can clear the mind and generate new energy. The combination of the two can make blood flow smoothly, brain meridians unobstructed, essence superior and marrow filling, blood ascending and nourishing the brain, promoting blood circulation, removing blood stasis, and awakening the mind.

This study obtained 25 compounds of Danshen Chuanxiong through OB and DL screening on the TCMSP platform, as well as searching for relevant literature to supplement characteristic compounds in traditional Chinese medicine. Among them, Danshen’s active ingredient, Danshentong IIA, had the highest activity and could act on 32 targets, while Ligusticum chuanxiong had the highest activity in Chuanxiong with 27 targets. Research has shown that tanshinone IIA can inhibit the proliferation of microglia, further reduce the expression of inflammatory cytokines IL-1 β and TNF – α, and inhibit excessive phosphorylation of Tau protein to combat neuronal apoptosis, thereby exerting a therapeutic effect on AD. Research has found that ligustrazine can regulate the expression of Bcl-2, Bax, and caspase-3 by acting on the PI3K/Akt pathway, thereby antagonizing A β 25-35-induced cell apoptosis.

The GO enrichment analysis results indicate that the targets of the “Danshen Chuanxiong” drug are distributed in various cellular components, mainly in the cytoplasm, and participate in biological processes through various binding modes, including inflammatory response, apoptosis process, oxidative stress response, aging, memory, MAPK cascade response, positive regulation of neuronal apoptosis process, regulation of inflammatory response, active regulation of leukocyte activation, nervous system development, etc. Research shows that inflammatory reactions in the brain are an important link in the onset of AD. Research has confirmed that apoptosis and oxidative stress response play extremely important roles in the occurrence and development of Alzheimer’s disease. Suggesting that “Danshen Chuanxiong” medicine may exert its therapeutic effect on AD through multiple targets and pathways. In order to validate the analysis results of network pharmacology, this study selected Danshen Chuanxiong to experimentally verify the cell survival rate, inflammatory response, and apoptosis rate of PC12 cells induced by A β 25-35. The results showed that Danshen Chuanxiong can significantly improve the cell survival rate of PC12 cells, inhibit the inflammatory response of PC12 cells, and reduce the apoptosis rate of PC12 cells, exerting a therapeutic effect on AD.

KEGG enrichment pathway analysis showed that the MAPK signaling pathway ranked first, followed by the NF kB signaling pathway. The MAPK signaling pathway in mammalian cells is mainly composed of the p38 signaling pathway, ERK signaling pathway, and JNK signaling pathway. Research has found that the expression level of p38 in brain tissue is highly correlated with the severity of dementia. Research has found that the p38MAPK pathway can participate in the process of neuronal cell apoptosis by promoting Bax translocation and activating Caspase-3 activity. Research has shown that inhibiting the p38MAPK pathway can suppress apoptosis of brain tissue cells induced by A β 1-42 in AD model rats. The NF kB signaling pathway can participate in the signal transduction processes of inflammation, neuronal apoptosis, and various gene regulation in the nervous system. Research has shown that p38MAPK is closely related to NF kB, and activated p38MAPK can induce the expression of inflammatory cytokines to activate NF kB. If the expression of p38MAPK is inhibited, the activation of NF kB will also be inhibited. Danshen Chuanxiong may inhibit inflammation and cell apoptosis by regulating the p38MAPK/NF kB signaling pathway, thereby delaying the occurrence and development of AD.

This study selected Danshen Chuanxiong to experimentally verify the network pharmacology research results of its regulatory effect on the p38MAPK/NF kB signaling pathway induced by A β 25-35 in PC12 cells. The experimental results indicate that Danshen Chuanxiong can significantly downregulate the p38MAPK/NF kB signaling pathway, ultimately inhibiting the development of AD.

This study applied network pharmacology to investigate the specific targets and molecular mechanisms of Danshen Chuanxiong in the treatment of Alzheimer’s disease. The results showed that Danshen Chuanxiong’s treatment of Alzheimer’s disease is mainly achieved through the synergistic effects of multiple components, targets, and pathways, providing important guidance for clinical research on the mechanism of Danshen Chuanxiong’s treatment of Alzheimer’s disease.