| BackgroundCancer-related fatigue(CRF)occurs widely in cancer patients,which seriously affects the patient’s quality of life and anti-cancer confidence,reduces the patient’s treatment compliance,and the survival rate.The risk factors of CRF are relatively complex.There are currently no clear clinical markers that can predict the occurrence of CRF and evaluate its therapeutic efficacy.The current modern treatments for CRF also have considerable limitations,so it is very important to clarify the clinical risk factors of CRF.Previous studies have shown that traditional Chinese medicine(TCM)has considerable efficacy in treating CRF,but the mechanism of TCM in interfering with CRF?This study then launched an in-depth discussion on these two issues.ObjectivesThrough clinical research,network pharmacology analysis and animal experiments,to evaluate the clinical risk factors that affect the occurrence of CRF after chemotherapy.The mechanism of TCM treating CRF and whether there is a certain connection between the TCM and risk factors were also discussed.Methods1.Clinical research part:Enrolled 271 cancer patients and divided them into 2 groups based on whether they were associated with CRF.The statistics index included staging,history of surgery,chemotherapy courses,radiotherapy,targeted therapy,immunotherapy,BMI,blood cell count,Biochemical and liver and kidney function,coagulation function,inflammatory index,glucose metabolism level.SPSS 22.0 statistical software was used to conduct single-factor and multi-factor analysis to determine independent risk factors,and to draw a receiver operating characteristic curve(ROC)of the model’s prediction rate,evaluate the validity of the prediction model,and analyze the criticality of the factor Value,sensitivity,specificity and Youden index.2.Network pharmacology part:This study Searched the randomized controlled trials of TCM in the treatment of CRF through the literature database,and screened the Chinese medicines with a frequency of≥3.To explore the treatment mechanism,we screened the active compounds and targets of traditional Chinese medicines in the treatment of CRF through the Chinese medicine database and disease database,and draw charts to analyze the interaction relationship.In order to analyze the biological process and signal pathway conduction process of traditional Chinese medicine acting on CRF,and then to further clarify the mechanism of drug action,the target in the network was enriched with GO and KEGG pathway,and the target and pathway were verified by molecular docking.This part can preliminarily explain the potential targets and pathway mechanisms of Chinese medicine for CRF.3.Animal test part:A total of 30 Balb/c mouse models were divided into 5 groups with6 mice in each group.Normal control group received intraperitoneal injection of corresponding volume of normal saline;intraperitoneal metastasis model group received intraperitoneal injection of corresponding volume of normal saline;low-dose Astragalus-Atractylodes macrocephala decoction,3.0g/kg(0.75ml/kg),gavage,Once a day for 10 days;Astragalus-Atractylodes in the middle dose group was given Astragalus-Atractylodes decoction,6.0g/kg(1.5ml/kg),gavage,once a day for 10 days;Astragalus-Atractylodes high dose The group was given Astragalus-Atractylodes decoction,12.0g/kg(3ml/kg),gavage once a day for 10 days;the abdominal metastasis model was inoculated with ct-26 cells and ct-26 human colon cancer cell lines were cultured in complete In the medium(high glucose DMEM medium+10%fetal bovine serum,p H 7.4),place it in a 37°C,5%CO2 incubator,change the medium once every 3 days,and when the cells are 70-80%confluent,The cells were digested with 0.25%trypsin for passage.All Babl/c mice were placed in a swimming box(height 30cm,diameter 25cm)before tumor implantation for adaptive swimming training for 2 days,5 min/day,and the water temperature was 25℃±1℃.The cultured ct-26 cells were digested and centrifuged,then mixed with matrigel in equal proportions to make a suspension of about 1×10~7cells/ml.Disinfect the skin of the right lower abdomen of the mouse and inoculate 0.2ml(about1×10~6tumor cells)into the abdominal cavity.After that we observed and recorded the general condition,body weight,eating status,hair color,tumor volume,liver and gastrocnemius muscle weight of mice daily,at the same time we performed exhaustive swimming test on mice,and detected the blood samples of mice by ELISA,flow cytometry,etc.Alanine aminotransferase(ALT),aspartate aminotransferase(AST),urea(BUN),creatinine(CR),uric acid(UA),glucose(GLU),fructosamine(Fru),lactate dehydrogenase(LDH),C-reactive protein(CRP),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),superoxide dismutase(SOD)were tested.This part can further explain the deeper mechanism of TCM treatment of CRF.Results1.Part of the clinical research showed that creatinine,interleukin-6,and C-reactive protein were independent risk factors for fatigue in patients with malignant tumors,and the result predicted a correct percentage of 80.3%,which was in good fit with the reality.By analyzing the ROC curve,it was found that when the CRP level in the serum of patients with malignant tumor exceeded 6.45 mg/L or IL-6 exceeded 9.06 pg/m L,about 80%of the patients could develop CRF,with good sensitivity and statistical significance,which had clinical predictive value.2.The results of network pharmacology showed that this study obtained a total of 540documents through database search.After screening,24 randomized controlled trials(RCTs)of Chinese medicine treatment of CRF were finally included.Through frequency statistics,this study found the commonly used Chinese medicine include Atractylodes macrocephala,Astragalus,Tangerine peel,Poria,Licorice,Malt and Ligustrum lucidum.The results of Network pharmacology showed that the main active compounds of these 7 traditional Chinese medicines include quercetin,kaempferol and luteolin,and a total of 447 Chinese medicine-related targets were obtained,which were compared with the collected 5003cancer-related fatigue targets.After matching and drawing Venn diagram,328 traditional Chinese medicine targets that interact with cancer-induced fatigue were finally obtained.Targets with higher degrees include AKT1,TP53,MAPK1,APP,IL-6,INS,STAT3,JUN,VEGFA,and MAPK8.Through the component target network diagram,it is also verified that traditional Chinese medicine has a multi-component,multi-target,and integrated adjustment method for treating cancer-related fatigue.Subsequently,this study conducted GO function enrichment analysis and KEGG pathway enrichment analysis through related databases,and selected items with higher values.Among them,biological processes included drug response,positive regulation of RNA polymerase II promoter transcription,and signal transduction.Negative regulation of apoptosis process,etc.;cell location includes plasma membrane,cytoplasm,nucleus,cytosol,components of plasma membrane,etc.;molecular functions include protein binding,protein homodimerization activity,enzyme binding,ATP binding,etc.The KEGG pathway enrichment analysis showed 10related pathways with Corrected P value≤0.01 and gene frequency>0.5%,including metabolic pathways,cytokine-cytokine receptor pathways,and cancer signaling pathways,PI3K-Akt signaling pathway,mitogen-activated protein kinase signaling pathway,RAS signaling pathway,etc.3.The result of animal experiment showed that:(1)General conditions of tumor-bearing mice:Before administration,there was no significant difference in the general conditions of mice in each group,such as mental state,activity,skin appearance,and diet.The surface of the mouse skin is delicate and shiny without wrinkles,the skin is thin,moist and elastic,lively,sensitive to food,and quick to escape.During the experiment,the mice in the normal group showed no obvious changes.The mice in the other groups gradually showed dull skin,slow movement,inactivity,and loss of appetite.Before the intervention,there was no significant difference in the body weight of each group.The normal group and the model group did not change significantly before and after the intervention,while the low-dose group(F=3.699,p=0.034)and the middle-dose group(F=13.4,P<0.001),the weight of the high-dose group(F=15.58,p<0.001)increased,and there was statistical significance.Beginning on the 6th day of intervention,the weight gain of the low-dose group and the high-dose group(F=4.28,p=0.012)was statistically significant,and the weight increase of the high-dose group was higher than that of the low-dose group and the normal group,and the weight of the low-dose group The increase is higher than the normal group.On the 9th day of intervention,the weight of the high-dose group(F=9.69,p<0.001)increased significantly,which was higher than that of the low-dose group,model group and normal group,which was statistically significant.(2)Exhaustive swimming experiment of tumor-bearing mice:There was no significant difference in swimming time before the intervention in each group.After the intervention,the exhaustive swimming time of the low-dose group on the 6th and 9th day was significantly lower than that on the 3rd day(F=10.127,p<0.001),and there was statistical significance,while there was no significant change in other groups before and after intervention.In the low-dose group,the swimming time to exhaustion on the third day of intervention was significantly higher than that of other groups(F=2.806,p=0.047),which was statistically significant.The exhaustive swimming time of the high-dose group on the9th day of intervention was longer than that of the other groups except the normal group(F=3.393,p=0.028),and it was statistically significant.(3)Test results of serum samples from tumor-bearing mice:This study found that the levels of IL-6 in each group using traditional Chinese medicine were lower than the model group,but higher than the normal group.The IL-6 level of mice in the low-dose group decreased most significantly,and the difference with the model group was statistically significant;the TNF-αlevel was statistically different between the groups(P=0.008),the low-dose group and the normal group There is a difference between the two groups(P=0.034).The levels of TNF-αin each group of mice using traditional Chinese medicine are lower than those in the model group,but the level of TNF-αin the low-dose group has the most significant decline,and is compared with that of the normal group.The difference is statistically significant;there is a statistical difference between the CRP levels(P=0.007),and there is a difference between the middle dose group and the normal group(P=0.022),except for the difference between the normal group and the middle dose group Except for the difference in CRP levels,the differences in the other groups are not significant;there is a statistical difference between the AST levels(P=0.02),and there is a difference between the high-dose group and the normal group(P=0.03),except for the normal group The AST level was the lowest,and the difference was statistically significant from the high-dose group of mice;the GLU level of each group was statistically different(P=0.048),and there was a difference between the high-dose group and the middle-dose group(P=0.029),the difference in blood glucose between the model group and the traditional Chinese medicine intervention group is not obvious,only in the high-dose group and the middle-dose group of mice;there are statistical differences in the Fru level between the groups(P=0.006),suggesting At least two groups are statistically different.Pairwise comparison shows that there is a difference between the middle dose group and the normal group(P=0.008).The normal group has a lower Fru level than the other groups,and the difference between the other groups is not significant,Only the difference between the middle-dose group and the normal group is statistically significant;the UA levels of each group are statistically different(P=0.025),and then through pairwise comparison,it is found that the high-dose group and the normal group are between the two groups There are differences(P=0.018).The UA level of the normal group is lower than that of the other groups,and the difference between the other groups is not obvious.Only the difference between the high-dose group and the normal group is statistically significant.(4)Weight inhibition of tumor-bearing mice:There was no significant difference in the abdominal circumference of mice in each group before intervention.On the third day after intervention,the abdominal circumference of the model group increased significantly compared with other groups(F=5.416,P=0.004)And the abdominal circumference of this group showed a rising trend on the 3rd,6th and 9th day of intervention,which was statistically different from other groups.In the normal group,low-dose group,middle-dose group,and high-dose group,the abdominal circumference of mice in each group did not increase significantly on the 3rd and 6th day after the intervention,but on the 9th day after the intervention,the abdominal circumference of each group appeared to varying degrees.Except for the normal group,the increase in tumor-bearing mice in each group was larger,which was statistically significant compared with before the intervention;after the intervention,there were differences in tumor weight in at least two groups of mice(P=0.043).Through pairwise comparison,the study found that the tumor weight of the mice in the middle-dose group was significantly smaller than that of the model group(P=0.044).The tumor weights of the mice after using Chinese medicine decreased to varying degrees,and the middle-dose group was the most obvious.By calculating the T/C value,it was also found that the T/C value of mice in the middle dose group was the smallest(51%)and the tumor inhibition rate was the highest.(5)Energy metabolism of tumor-bearing mice:After the intervention,there was no statistically significant difference in the weight of the liver and gastrocnemius muscle of each group of mice(P=0.19,P=0.12);the liver SOD activity was statistically significant(P=0.03),That is,the SOD activity of the low-dose,high-dose group,and model group is equivalent,but higher than the normal group and the middle-dose group,indicating that the middle-dose group has the least amount of oxygen free radicals,liver cell damage,and SOD activity.There was no significant difference in liver LD content,liver tissue homogenate protein content,gastrocnemius homogenate protein content,muscle SOD activity,and muscle LD content after intervention.However,the protein content of liver and muscle homogenate,the mouse group of each dose compared with the model group,has an increasing trend;the muscle SOD activity of each mouse using Chinese medicine has a downward trend compared with the model group;while the LD content of the liver and gastrocnemius of each group.It can be seen that there is an upward trend in each mouse group using traditional Chinese medicine compared with the model group.Conclusion1.Interleukin-6 and C-reactive protein are important independent risk factors for CRF patients.Clinically,treating cancer patients according inflammatory response may improve the symptoms of fatigue in those patients;2.TCM is often used to invigorate qi Spleen-invigorating drugs for the CRF petients,and the common Chinese herb include Atractylodes,Astragalus,Tangerine Peel,Poria,Licorice,Malt and Ligustrum lucidum.Network pharmacological analysis showed that the mechanism mainly involved the regulation of metabolic pathways,cytokine pathways,and cancer signaling pathways through multiple ingredients and multiple targets to exert anti-CRF effects.3.Animal studies have shown that Astragalus and Atractylodes macrocephala can improve fatigue in mice with metastatic tumors in the abdominal cavity,increase weight,and inhibit tumor growth.Its efficacy may be related to reducing inflammatory cytokines in mice,maintaining glycogen reserves,and reducing oxidation.In addition,the efficacy is related to the regulation of energy metabolism. |
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