The compatibility rules and systematic pharmacological mechanism analysis of turmeric in blood regulating formulas
Chinese medicine turmeric is the dried rhizome of Curcuma longa L., a plant in the ginger family. It has a unique aroma, bitter and pungent taste, and can regulate the spleen and liver meridians. It has the effects of breaking blood, promoting qi circulation, and relieving pain. Turmeric is combined with traditional Chinese medicine in prescriptions, starting from the Wu Bi Tang in the “Tai Ping Hui Min He Zhi Ju Fang”. It is used as an equivalent for regulating blood circulation, relieving pain, and treating wind cold dampness evil, as well as leaving skin on the guest side. Later generations of medical practitioners often used turmeric as a blood regulating medicinal herb to treat internal medicine rheumatism, gynecology, bone injuries, miscellaneous diseases, and other fields. In addition, turmeric is often used as an ethnic medicine or health food in Iran, India, Thailand and other places to treat skin diseases, infections, mental illnesses, etc. Given that turmeric can have therapeutic effects on various diseases, in recent years, many scholars at home and abroad have made unremitting efforts to study the molecular pharmacological mechanisms of turmeric. Eleonora et al. reviewed the modern pharmacological mechanisms of turmeric and found that it has effects on improving metabolic disorders, chronic diseases, regulating functions, antioxidation, and health care. Deguchi et al. found that turmeric and its main active ingredient curcumin have high safety and can mediate multiple targets and pathways to exert anti-inflammatory and anti-tumor effects. Research on the pharmacological effects of turmeric in China has mainly focused on its anti-tumor, anti-inflammatory, hypoglycemic, and cardiovascular effects.

Although current research has revealed many pharmacological mechanisms of turmeric, it is mostly limited to the study of turmeric as a single medicinal herb or its components, and there are few researchers who have studied its compatibility rules and the synergistic pharmacological mechanisms of compound formulations. As a component of traditional Chinese medicine prescriptions for treating diseases, the efficacy of turmeric is inevitably influenced by the interaction of compatible drugs. The theory of traditional Chinese medicine holds that “medicine has its own unique strengths, and prescriptions have the beauty of combining with others”. The functions of the components of the prescription, such as Jun, Chen, Zuo, and Shi, are different. In the “Shennong Bencao Jing”, there are discussions such as “Xiang Xu and Xiang Jiang represent good, while those who oppose or oppose each other should not be used”. Therefore, exploring the pharmacological effects of turmeric should not be limited to its own research, but should also involve a comprehensive analysis of its formula compatibility. The advancement of data mining technology has provided new methods for studying the association rules of traditional Chinese medicine compatibility. Among them, Apriori association algorithm and generalized rule induction (GRI) algorithm are commonly used simple association rule algorithms, which can effectively mine the core drug pairs and prescriptions with strong correlation in prescriptions. This not only simplifies the number of ingredients in prescriptions, but also retains the beauty of traditional Chinese medicine compatibility. It is widely used in the discovery of traditional Chinese medicine compatibility rules and core formulas. For example, Tang Qisheng et al. used the Generalized Rule Induction (GRI) association rule algorithm for data analysis and discovered traditional Chinese medicine core formulas for treating anxiety, such as Chaihu Yujin and Suanzaoren Shenglonggu. This study is based on the principle of database mining technology, using GRI algorithm, Arrowsmith knowledge discovery tool, bioinformatics database and other methods to investigate the compatibility rules and pharmacological mechanisms of traditional Chinese medicine turmeric in blood regulating formulas, in order to provide theoretical support and technical clues for the development of new generation turmeric products.

Blood regulating agents are a type of traditional Chinese medicine formula that has the effects of regulating blood flow, dispersing blood stasis, cooling blood, and relieving pain. Modern medicine is commonly used to treat traumatic swelling and pain, gynecological inflammation, cardiovascular and cerebrovascular diseases, tumors, and cancers. The core compatibility of turmeric found in this study, “turmeric Angelica sinensis chuanxiong”, in which chuanxiong belongs to the category of promoting blood circulation and removing blood stasis medicine, while Angelica belongs to the category of promoting blood circulation and nourishing blood medicine. The compatibility of turmeric with Ligusticum chuanxiong and Angelica sinensis has a good theoretical basis for traditional Chinese medicine compatibility. Chuanxiong is a blood qi medicine that also has the effect of promoting qi circulation and relieving pain. Qi circulation promotes blood flow, thus enhancing the blood circulation and stasis removing ability of turmeric and angelica. Angelica sinensis is a holy herb for nourishing blood, which can dispel blood stasis and promote rejuvenation. It is combined with turmeric and Ligusticum chuanxiong to dispel evil, strengthen the body, and promote both attack and nourishment. The ancient classic formula “Siwu Tang” has the usage of combining Angelica sinensis with Chuanxiong. Li et al. found that the combination of Angelica sinensis and Ligusticum chuanxiong has better therapeutic effects on blood stasis rats than Angelica sinensis or Ligusticum chuanxiong alone. Deng et al. summarized that through data mining of ancient and modern famous formulas, it was found that turmeric has a therapeutic effect on brain diseases and is best combined with blood activating and stasis removing drugs such as Angelica sinensis and Chuanxiong. This is consistent with the data mining conclusions of this study. In summary, it indicates that Angelica sinensis, Ligusticum chuanxiong, and Curcuma longa have a synergistic effect, but further experimental analysis is needed to determine the mechanism of the synergistic effect of Curcuma longa with Angelica sinensis and Ligusticum chuanxiong.

This study first used the Arrowsmith knowledge discovery tool to discover that “turmeric Angelica sinensis chuanxiong” can act in combination on 13 targets including AKT, TLR4, caspase-3, PI3K, mTOR, p38 MAPK, VEGF, iNOS, Nrf2, BDNF, NF – κ B, Bcl-2, Bax, involving 27 related genes. PPI and pathway enrichment show that “Curcuma longa Angelica chuanxiong” can regulate PI3K/Akt, MAPK, Toll like receptors, T cell receptors, EGFR, VEGF, Apop tosis, HIF-1 and other signaling pathways through hub genes such as AKT1, PIK3CA, RELA, NFKB1, MAPK14, VEGFA, etc. It is predicted that it may have potential pharmacological effects such as enhancing immunity, anti-inflammatory, improving microcirculation, protecting brain nerves, anti-tumor, and anti-tumor angiogenesis.

Liu et al. found that turmeric can inhibit the AKT/mTOR pathway, leading to apoptosis of ovarian cancer cell line SK-OV-3. Ashrafizadeh et al. found that turmeric has multiple effects by targeting the Nrf2 signaling pathway, including antioxidant, antibacterial, anti-tumor, and anti-inflammatory effects. In addition, studies have shown that Ligusticum chuanxiong and Angelica sinensis can also exert anti-tumor effects. Shen et al. used RT qPCR and Western blotting analysis to show that Ligusticum chuanxiong can significantly reduce the expression and activity of Akt, increase the activity of caspase-3, and also exhibit anti-tumor effects on various types of cancer. Angelica sinensis can induce gastric cancer cell death by inhibiting mTOR protein and significantly prolong the survival rate of cancer patients. Angelica also affects the expression of Bcl-2 related X protein (Bax), Bcl-2 and caspase-3, a variety of apoptosis factors, and promotes the apoptosis of human breast cancer cells. Therefore, it has been proved to be a promising therapeutic agent for breast cancer. In addition, the formula of “turmeric Angelica sinensis chuanxiong” may have anti-tumor angiogenesis effects. ELISA analysis shows that turmeric can inhibit the secretion of VEGF in tumor cells, further suppressing tumor angiogenesis. Chen et al. found that Angelica sinensis can prevent cancer cell proliferation and angiogenesis by inhibiting the PI3K/AKT/mTOR signaling pathway and downregulating the expression of HIF-1 α and VEGF. These studies indicate that the formula of “turmeric Angelica sinensis chuanxiong” can exert anti-tumor effects. Our results are basically consistent with the above research.

In addition, this study speculates that the formula of “turmeric Angelica sinensis chuanxiong” can also exhibit anti-inflammatory, neuroprotective, and microcirculation improving effects. Zhu et al. used immunohistochemistry, enzyme-linked immunosorbent assay, and Western blotting to demonstrate that turmeric can alleviate acute inflammatory injury in experimental traumatic brain injury by inhibiting the TLR4/MyD88/NF – κ B signaling pathway. Studies have shown that Angelica sinensis can upregulate the expression of TLR4 protein and has pharmacological effects of anti-inflammatory and analgesic. Chuanxiong can reduce the expression of inflammatory cytokines and adhesion molecules on vascular endothelial cells by inhibiting the p38 MAPK and NF – κ B signaling pathways. These studies confirm the hypothesis of the anti-inflammatory effect of the formula of “turmeric Angelica sinensis chuanxiong”. Wu et al. found that turmeric can play a beneficial role in ischemic brain injury by upregulating the expression of caspase 3 and Bcl-2, downregulating the expression of Bax, and promoting the activation of the PI3K/AKT pathway. Hurley et al. found that turmeric can lead to a dose-dependent increase in BDNF in the hippocampus and has a neuroprotective effect. Cheng et al. found that Angelica sinensis can protect patients from cerebral infarction and improve neurological damage by downregulating caspase 3 and upregulating p-p38 MAPK, hypoxia inducible factor (HIF-1), and vascular endothelial growth factor-A (VEGF-A). Chen et al. found that Danggui Sini Tang can effectively protect the microvascular endothelial cells of pancreatic islets through the PI3K/Akt/iNOS pathway. Our results are consistent with the above studies, demonstrating the pharmacological effects of the formula of “turmeric angelica chuanxiong” in protecting the brain nerves and improving microcirculation.

Finally, this study used MTT, Elisa, and Western blotting experiments to find that the formula of “turmeric Angelica sinensis chuanxiong” can inhibit the proliferation of human lung cancer A549 cells and downregulate the expression of p-EGFR, p-PI3K, p-Akt, VEGF, and HIF-1 α in A549 cells. These experiments demonstrate that the formula of “turmeric Angelica sinensis chuanxiong” does indeed have anti-cancer and anti-tumor angiogenesis effects in vitro, proving that data mining information is reliable and effective, and can predict certain pharmacological mechanisms of traditional Chinese medicine. The results of this study provide methodological references for the development of traditional Chinese medicine and the study of pharmacological mechanisms.