Study On The Effect Of Bushentongluo Decoction On Central Sensitization Of Bone Cancer Pain Based On ERK/c-Fos Signaling Pathway

Background of the studyBone metastatic cancer pain is a continuous and incurable disease caused by the tumor’s invasion of the bone and its surrounding soft tissues,which leads to bone cancer pain.And it poses a huge challenge to relieve the pain of patients and has an adverse impact on the mental health of patients.Pain sensitization,whose pain nature is enhanced pain sensitivity and enhanced pain response,is a remarkable feature of bone metastatic cancer pain sensitization.The drug resistance rate of the three-step analgesic therapy recommended by the World Health Organization(WHO)is very high,reaching 37%to 57%,and the long-term use of opioid drugs will aggravate pain sensitization.Professor Fang Zhijun,the supervisor,combines Chinese and Western medicine,increases efficacy and reduces toxicity,and relieves pain.In tumor treatment,it is considered that the pathogenesis of bone metastatic cancer pain is mainly the deficiency of kidney essence and the loss of nourishment of bone marrow.In the treatment of bone metastatic cancer pain,Eucommia ulmoides,Drynaria fortunei,Curcuma aromatica,Rhodiola rosea,Scorpio,and Aspongopus are often used in combination.The combination of multiple drugs mostly has the functions of tonifying the kidney and warming yang,strengthening the bone and relieving pain,which can effectively relieve bone metastatic cancer pain.Research objectivesThrough network pharmacology,obtain the target information of bone metastatic cancer pain and the Bushen Tongluo decoction,construct the "active ingredienttarget" network and the target protein interaction network,and predict the action targets and pathways.Use the ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method and the negative ion multiple reaction monitoring(MRM)mode for content determination and analysis to determine that the Bushen Tongluo decoction has components with potential to relieve bone metastatic cancer pain sensitization.Through experiments,verify the impact of the Bushen Tongluo decoction on the relevant signaling pathways at the spinal dorsal horn.Research methods1.Inpatients in Suzhou Hospital of Integrated Traditional Chinese and Western Medicine from 2021 to 2023 were screened according to the diagnosis and inclusion and exclusion criteria.Enter the patient’s name,gender,age,chief complaint,physical examination,name of traditional Chinese medicine disease,syndrome type,prescription and medication situation.Record the patient’s Visual Analogue Scale(VAS)score for pain intensity,Generalized Anxiety Disorder(GAD)score,and the depression module score of the Patient Health Questionnaire(PHQ-9)on file,and conduct the analysis and comparison of the clinical efficacy evaluation of the Bushen Tongluo decoction.2.The Traditional Chinese Medicine Systems Pharmacology(TCMSP)database is used to search for the components and targets of the Bushen Tongluo decoction.In addition,we used the keyword "bone metastatic cancer pain" to explore the GeneCards database and the Online Mendelian Inheritance in Man(OMIM)database.Six kinds of medicinal materials such as Eucommia ulmoides,Drynaria fortunei,Scorpio,Aspongopus,Curcuma aromatica,and Rhodiola rosea were selected as the research objects.Input the screened targets of the active ingredients of the Bushen Tongluo decoction and the targets of bone metastatic cancer pain into the Venn diagram making software to identify the common targets.Then input these common targets into the String database to construct a PPI network.The PPI network is further imported into Cytoscape for analysis.Conduct Gene Ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis on the common targets of the Bushen Tongluo decoction and bone metastatic cancer pain to obtain the main active ingredients and targets of the Bushen Tongluo decoction.Secondly,the ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)technology was used to determine the active ingredients of the Bushen Tongluo decoction,and the UPLC-MS/MS pharmacodynamic component analysis method was used to identify the main compounds therein.The results showed that a total of 17 main components were found.Among them,salidroside in Rhodiola rosea and curcumin in Curcuma aromatica were confirmed to have the effect of anti-cancer pain,and protocatechuic acid and naringin in Drynaria fortunei could inhibit bone cell damage and protect bone quality.3.Prepare ascites tumor cells with Walker-256 breast cancer cells,and establish an animal model of bone metastatic cancer pain after tibial inoculation.Use the Bushen Tongluo decoction to intervene in the bone metastatic cancer pain model,use q-PCR to detect the mRNA content of ERK/c-Fos-related pathway proteins in the spinal dorsal horn;use Western blot to detect the content of ERK/c-Fos signal pathway-related proteins;use enzyme-linked immunosorbent assay(ELISA)to detect the amounts of IL-6,IL-1 β,and TNF-α in the abdominal aortic serum to clarify the inhibitory effect of the Bushen Tongluo decoction on the central sensitization of bone metastatic cancer pain.Study results1.The research results show that the VAS score of the patients in the experimental group at the end of the treatment was significantly lower than that of the control group,and the difference was statistically significant(P<0.05).The GAD score and PHQ-9 score of the patients in the experimental group at the end of the treatment were also significantly lower than those of the control group,and the differences were both statistically significant(P<0.05).No obvious adverse reactions occurred in the patients of the experimental group during the treatment process.From the comprehensive research results,it can be known that the Bushen Tongluo decoction has a significant curative effect on bone metastatic cancer pain,can effectively relieve the pain perception of patients,and reduce anxiety and depression symptoms.This decoction has good safety and tolerance and can be used as an effective traditional Chinese medicine decoction for treating bone metastatic cancer pain.2.The research selected six kinds of medicinal materials such as Eucommia ulmoides,Drynaria fortunei,Scorpio,Aspongopus,Curcuma aromatica,and Rhodiola rosea as the objects.Through the network pharmacology method,the action target network of the Bushen Tongluo decoction was constructed.The research found that this decoction is mainly involved in regulating biological processes such as the immune system,inflammatory response,and ERK/c-Fos signal transduction.The detection by UPLC-MS/MS technology shows that salidroside in Rhodiola rosea,curcumin in Curcuma aromatica,protocatechuic acid in Drynaria fortunei,and naringin can play a role in inhibiting bone destruction and relieving cancer pain.In the market,these components show very high pharmacological activity and may become the research and development direction of new drugs.3.Bone metastatic cancer pain is a refractory pain,which seriously affects the quality of life and mental health of patients.This study aims to explore the analgesic effect of the Bushen Tongluo decoction on rats with bone metastatic cancer pain and its influence on the expression of related inflammatory factors and ERK/c-Fos protein pathways.In this study,Walker-256 cancer cells were injected into the tibia of rats to induce bone metastatic cancer pain in rats.The expression of inflammatory factors in serum was determined by enzyme-linked immunosorbent assay.Western blot and q-PCR were used to detect ERK and c-Fos in the spinal dorsal horn.Eventually,mechanical allodynia and thermal hyperalgesia were observed in rats with bone metastatic cancer pain.The activation of the ERK/c-Fos pathway was observed in the spinal dorsal horn,and the expression of IL-1β,IL-6,and TNF-α in the serum increased.The Bushen Tongluo decoction alleviates bone metastatic cancer pain and reduces ERK/c-Fos and inflammatory cytokines.Conclusion of the study1.The Bushen Tongluo decoction combined with oxycodone has a significant curative effect on the treatment of bone metastatic cancer pain.Compared with the single use of oxycodone for treatment,it can effectively relieve the pain perception of patients and reduce anxiety and depression symptoms.2.The Bushen Tongluo decoction can play a role by regulating multiple biological processes such as the immune system,inflammatory response,and signal transduction.The active ingredients therein include salidroside,curcumin,protocatechuic acid,naringin,etc.,which can significantly relieve the symptoms of bone metastatic cancer pain.The research found that the Bushen Tongluo decoction may play a therapeutic role through the ERK/c-Fos signal pathway,providing a potential drug choice for treating bone metastatic cancer pain and providing a theoretical basis for in-depth exploration of its treatment mechanism and the development of new drugs.3.The evidence we provided shows that the Bushen Tongluo decoction has a potential beneficial effect on bone metastatic cancer pain and can effectively reduce the allodynia and hyperalgesia of rats with bone metastatic cancer pain.This effect may be attributed to the down-regulation of the ERK/c-Fos signal in the spinal dorsal horn and the serum inflammatory cytokines IL-1β,IL-6,and TNF-α.