Study On The Therapeutic Effect And Mechanism Of Zhengyuan Capsule In The Prevention And Treatment Of Cancer-related Fatigue

ObjectiveBy analyzing the clinical data and the establishment of cancer-related fatigue(CRF)in nude mice model of lung cancer,we evaluated the effect of Zhengyuan capsule in the prevention of CRF caused by tumor and chemotherapy,and clarify the mechanism of the capsule in alleviating CRF.Methods1.Clinical research:To explore the theoretical basis for the prevention and treatment of cancer related fatigue in traditional Chinese medicine,we used the Piper fatigue revised scale,self-made TCM syndrome differentiation combining with our previous research results.2.Animal experiment:Establishment of nude mice model of A549 lung adenocarcinoma transplantation tumor.20 nude mice were randomly selected as the normal control group(NC),and the other 160 were inoculated with A549 cells.After inoculation,when the tumor mass was 50-100mm3 mm3(seventh days after inoculation),they were randomly divided into 8 groups,with 20 in each group.All mice were divided into 9 groups:normal control group(NC),cancer model(TC)group,low dose of Zhengyuan capsule(ZD)dose group,Zhengyuan capsule(ZZ)group,high dose of Zhengyuan capsule(ZG)group,cisplatin(P)group,low dose cisplatin + Inpac capsule(LD)dose group,cisplatin + positive element capsule(LZ)group,high dose cisplatin + Inpac capsule(LG)group.The traditional Chinese medicine(Z)group included three subgroups(ZD,ZZ,and ZG),and the combination(L)group consisted of three subgroups(LD,LZ,and LG).The reference drug dose equivalent formula,set the Zhengyuan capsule low dose group was administered at a dose of 0.45g/kg,medium dose group of 0.9g/kg(human equivalent dose),high dose group 1.8g/kg orally,1 times a day.The dose of cisplatin was 5 mg/kg,intraperitoneal injection,once every 3 days.After grouped(A549 cells were inoculated seventh days),to start the drug,once a day,for 21 consecutive days.Normal control group and cancer model group were given the corresponding volume of normal saline.In this study,A549 lung adenocarcinoma in nude mice was used as the research object.To clarify the effect of Zhengyuan capsule in the treatment of CRF mice,we observed the swimming time,open field test,the immobility time in tail suspension of the CRF mice.And flow cytometry,ELISA,Wright Giemsa staining,cytokine antibody array technology were used to test the peripheral blood cell count,bone marrow smear,cytokines,corticosrone,T3,T4,liver glycogen.Through this experiment,we intended to clarify the mechanism of Zhenyuan capsule in the prevention of CRF,and to provide experimental basis for clinical application for Zhengyuan capsule in the treatmetnt of CRF.Result1.In clinical study,we intedned to clarify the relationship between viscera deficiency and fatigue score.We found the scores of fatigue were higher in patients with Spleen Yang deficiency and liver qi stagnation,the difference was statistically significant(P<0.05).Patients with kidney yang deficiency and kidney qi deficiency stress also had high scores of fatigue,which was statistically significant(P<0.001).2.Pharmacodynamics research of Zhengyuan Capsule on mice suffering from cancer related fatigue.(1)The general situation of the mice in each group was balanced before the intervention.After 3 weeks of intervention,the general condition of the mice from the relatively good order to poor condition were NC group,ZD group,ZZ group,ZG group,TC group,LZ group,LD group,LG group and P group.The weight of tumor bearing mice were,NC group>TC group(P<0.023),L group>P group(P<0.024).(2)Effect of Zhengyuan Capsule on tumor-bearing mice body fatigueIn exhaustive swimming time,different groups of tumor-bearing mice were balance before the intervention,but there was statistically significant differences at the 18th day of intervention.The swimming time of TC group was shorter than the NC group(P<0.001).And the swimming time of each dose group in Z group was longer than that of TC group(P<0.001).Also,the swimming time of each dose group in L group was significantly longer than that of P group,and the P values were,P=0.004(LD group vs TC group),P<0.001(LZ group vs TC group,LG group vs TC group).Swimming time of LG group prolonged significantly than in LD group(P<0.001)and LZ group(P=0.017).The results show that,liver glycogen in TC group decreased compared with NC group,but there was no statistical significance(P<0.360).And the liver glycogen in Z group increased compared with TC group(P<0.001),also the liver glycogen in L group increased highly than in P group(P<0.001).(3)Effect of Zhengyuan Capsule on tumor-bearing mice mental fatigue.The tail suspension test showed that,after the intervention,the suspension tail of TC group was longer than that of NC group(P=0.031).An the suspension tail of Z group was significantly shorter than that of TC group(P<0.001),also,L group was significantly shorter than that of P group(P<0.001),as well as the TC group(P<0.001).Open field test showed that,after the intervention,total movement distance(P=0,040),the moving distance of the peripheral lattice distance(P<0.039),moving average speed(P=0.040)in NC group presented rising trend compared with TC group.The central grid movement distance of NC group was significantly shorter than TC group(P<0.046).When the mice were intervened with Zhengyuan Capsule,the central grid movement distance of Z group was significantly shorter than TC group(P<0.001).We also found-that,after the intervention,total movement distance(P=0.018),the moving distance of the peripheral lattice distance(P=0.002),moving average speed(P=0.017)in L group increased compared with P group,and the central grid movement distance significantly decreased compared with P group(P<0.001).Rest time in each group of tumor bearing mice did not find a significant difference.Stool grain number of tumor bearing mice were most in Z group andleast in P group.(4)Inhibition of Zhengyuan Capsule on tumor growth in tumor bearing mice.Tumor weight in P group(P<0.001),L group(P<0.001),Z group(P<0.032)decreased compared with the TC group.Tumor inhibition rate of Z group,L group,P group were 26.9%,60%,57.1%,respectively.At the intervention of 19d,tumor volume of TC group increased compared with the P group,the difference of which was statistically significant(P=0.019).And tumor volume of TC group was increased compared with the L group,the difference of which was also statistically significant(P<0.001).3 Effect of Zhengyuan capsule in hematopoietic and immune function on CRF mice.The count of WBC,HGB and bone marrow nucleated cell count,L group>P group(P<0.001).No effect of Zhengyuan capsule on RBC or PLT count was found.Effects of Zhengyuan Capsule on serum cytokine levels.The EPO content in serum of Z group was high than TC group(P=0.049)and L group high than P group(P<0.038).The G-CSF content in serum was L group>P group(P<0.043).The IL-3 content in serum was LD group>P group(P<0.001),Z group>TC group,and the difference was statistically significant,respectively P<0.001(TC group vs ZD group)?P=0.001(TC group vs ZZ group)?P=0.025(TC group vs ZG group).And the IL-3 content in serum was ZD group>ZG group(P=0.003),LD group>LZ group(P<0.001),LG group>LZ group(P=0.024).The TNF-? content in serum,LZ group(P=0.013)?LG group(P=0.010)decreased compared with P group,ZG group decreased compared with TC group(P=0.004),ZG decreased compared with ZD group(P=0.05)and ZZ group(P=0.024),LD grooup increase compared with LZ group(P=0.026)and LG group(P=0.021).All Sub groups of L group decreased compared with P group and TC group in IL-6 content of serum(P<0.001).And ZZ group and ZG group decreased compared with TC group in IL-6 content of serum(P<0.001).Also ZZ group decreased compared with ZD group(P<0.001).More over LZ group decreased compared with LD group and LG group(P<0.001).It seemed that middle dose of Zhengyuan Capsule was better for decreasing IL-6 content of serum.The IL-2 content in serum,the levels of Z groups were higher than that in the TC group and the difference was statistically significant.The content of IL-2 rose with the increasing in the dose of Chinese medicine,and the P value respectively was P=0.013(TC group vs ZD group)?P<0.001(TC group vs ZZ group)?P<0.001(TC group vs ZG group).The IL-2 content of LZ group and LG group IL-2 higher than that in P group,and the difference was statistically significant.The higher the dose of traditional Chinese medicine,the greater the IL-2 rise,and the P value respectively was P<0.001(P group vs LZ group)?P<0.001(P group vs LG group).Spleen index of L group increased statistically significantly compared with P group,(P=0.034).4 Effects of Zhengyuan Capsule on endocrine function of CRF miceThe results showed that,in the serum level of T3,P group decreased compared with TC group(P=0.008),Z group increased compared with TC group(P<0.001),L group increased compared with P group(P<0.001),and all the differences were statistically significant.And in the serum level of T4,Z group increased compared with TC group(P=0.002).We did not find effects of Zhengyuan Capsule on serum corticosterone.Conclusion1.CRF closely relats with the spleen and kidney,and involves lung and liver.The pathogenesis of CRF may be spleen deficiency,kidney essence deficiency and phlegm and static blood binding together.Because Zhengyuan capsule can supplement qi and strengthening the spleen,kidney essence,and soft solid scattered knot,it can be aimed at the pathogenesis of CRF of.2.Zhenyuan capsule can prevent and treat cancer-related fatigue in mice.3.Improving the hematopoietic and immune function,may be the mechanism of the effects of Zhengyuan capsule in treating CRF.4.Improving the thyroid function,may be the mechanism of the effects-of Zhengyuan capsule in treating CRF.