Protective Effect And Mechanism Of Kangai Injection On Cancer-related Fatigue In Mice

Cancer-related fatigue（CRF）is one of the common symptoms of clinical malignancies,a disturbing,persistent physical,emotional and/or cognitive subjective fatigue caused by tumor or anti-tumor therapy.Feeling and feeling exhausted,and interfere with daily life and function.This feeling of fatigue is more difficult to relieve than the pain and nausea associated with cancer.Therefore,there is an urgent need to find drugs that can effectively treat cancer-induced fatigue to improve the quality of life of patients with cancer-induced fatigue.This experiment will investigate the role and mechanism of Kangai injection in protecting mice from cancer due to cell,molecular and overall levels.Protective effect and mechanism of Kangai injection on fatigue of tumor-bearing miceTwenty male Balb/c mice were randomly selected as a blank group,and the remaining 80 subcutaneous injections（sc）0.2 ml mouse colon cancer cell（C26）suspension containing 5×10⁵ cells to prepare a transplantable colon cancer model.According to the body weight,they were randomly divided into 4 groups:model group,Kangai injection small,medium and large dose groups,20 in each group.The blank group and the model group mice were given intraperitoneal injection（ip）with normal saline 10 mL/kg,and Kangai injection small,medium and high dose groups were given ip to Kangai injection 5,10,20 mL/kg,daily 1 The drug was administered continuously for 21 days.Weekly blood was taken from the tail vein to measure white blood cells（WBC）,red blood cells（RBC）and hemoglobin（Hb）;treadmill experiments,rotating rod experiments and climbing rod experiments were performed30 minutes after the administration of the 17th,19th and 21st days of the drug.Lactic acid（LA）content in whole blood before mouse climbing rod experiment,after climbing rod experiment and after 10 breaks in climbing rod experiment;Nitric oxide（NO）and superoxide in serum were measured in eyeball blood taken 1 hour after medicine Dismutase（SOD）,malondialdehyde（MDA）,lactate dehydrogenase（LDH）,blood urea nitrogen（BUN）;extract tumors,calculate tumor inhibition rate;weigh heart,liver,spleen,lung,kidney weight Calculate the organ coefficient;take the quadriceps and liver of the bone,measure the liver glycogen,muscle glycogen,Ca²⁺-Mg²⁺-ATPase;take the gastrocnemius muscle,detect the muscle atrophy box F gene（MAFbx）,muscle ring finger by qRT-PCR Gene 1（MuRF1）mRNA expression.Protective effect and mechanism of Kangai injection on depression in tumor-bearing miceTwenty-two male Balb/c mice were randomly selected as a blank group,and the remaining 100 sc were administered with 0.2 ml of C26 suspension containing 5×10⁵cells to prepare a transplanted colon cancer model,which was randomly divided into5 groups according to body weight:model Group,Kangai injection low,medium and high dose group and positive drug fluoxetine group,20 in each group.The blank group and the model group mice were given ip with normal saline 10 mL/kg,Kangai injection small,medium and large dose groups were given ip to Kangai injection 5,10,20 mL/kg,and the positive drug group was intragastric（ig）fluoxetine 10 mg/kg was administered once daily for 21 days.Forced swimming,tail suspension and autonomic activity test were performed 30 minutes after the administration of the 17th,19th and21st day of the drug.The blood in the eyeball was measured 1 hour after the 21st day of the drug to determine the interleukin-6（IL-6）and interleukin in the serum.1β（IL-1β）;hippocampus,preparation of homogenate,measurement of serotonin（5-HT）,norepinephrine（NE）,dopamine（DA）,acetylcholinesterase（AchE）;qRT-PCR detection of hippocampal IL-6,IL-1βmRNA expression;Western Blot determination of Bcl-2,BAX,caspase-3,caspase-9 expression;hematoxylin and eosin staining（HE）detection of hippocampal neuron histopathological changes.Results:Protective effect and mechanism of Kangai injection on fatigue of tumor-bearing miceCompared with the blank group,the running time,running distance,swinging time and climbing time of the model group were significantly shorter（P<0.001）.Compared with the model group,the running time and running distance of Kangai injection low-dose group were compared.There was no significant effect on the time of rotatable sticking and climbing time（P>0.05）.The running time,running distance,swinging time and climbing time were significantly prolonged in the middle and high dose groups（P<0.01）.Compared with the blank group,there was no significant change in WBC,RBC and Hb in peripheral blood on the 7th day after modeling in the model group（P>0.05）.On the 14th and 21st day after modeling,the mice in the model group were WBC and RBC and Hb were significantly decreased（P<0.01 or P<0.001）.Compared with the model group,the small,medium and high doses of Kangai injection significantly increased WBC and Hb in mice（P<0.05^P<0.001）.There was no significant effect on the number of RBCs（P>0.05）.Compared with the blank group,there was no significant change in the heart index,Spleen index,and Kidney index of the model group（P>0.05）,while the liver index and lung index.The index was significantly decreased（P<0.05）.Compared with the model group,the small,medium and large doses of Kangai injection had no significant effect on heart,lung,liver and spleen and kidney coefficients（P>0.05）.Compared with the model group,there was no significant change in tumor weight in the low-dose group of Kangai injection（P>0.05）,and the tumor weight was significantly decreased in the middle and high-dose groups（P<0.001）;The tumor inhibition rate of the high-dose group of Ai injection is more than 30%.Compared with the blank group,the NO content and SOD activity in the serum of the model group were significantly decreased（P<0.01）,and the MDA content was significantly increased（P<0.01）.Compared with the model group,Kangai injection small dose group could make The serum MDA content was significantly decreased（P<0.05）.The MDA content in serum was significantly decreased in the middle and high dose groups,and the NO content and SOD activity were significantly increased（P<0.05 or P<0.01）.Compared with the blank group,the BUN content in the serum of the model group was significantly increased（P<0.01）.Compared with the model group,the small and medium dose groups had no significant effect on the BUN in the serum of the tumor-bearing mice（P>0.05）.The high dose group of Kangai injection can significantly reduce the BUN content of tumor-bearing mice（P<0.01）.Compared with the blank group,the LA content of the model group was significantly increased before exercise,after exercise and after 10 minutes of exercise（P<0.05）.Compared with the model group,Kangai injection low-dose group before and after exercise LA content was not significantly affected after 10minutes of exercise（P>0.05）.The medium and high dose groups could significantly reduce LA content before,after,and 10 minutes after exercise（P<0.05 or P<0.01）..Compared with the blank group,the LDH activity of the model group was significantly increased（P<0.001）.Compared with the model group,the low-dose group of Kangai injection had no significant effect on the LDH activity of the mice（P>0.05）,medium and large doses.The LDH activity in the serum of the mice was significantly reduced（P<0.05 or P<0.01）.Compared with the blank group,the glycogen and muscle glycogen content of the model group were significantly decreased（P<0.01 or P<0.001）.Compared with the model group,Kangai injection low dose group was used to treat mouse glycogen and muscle.Glycogen content had no significant effect（P>0.05）;medium and high dose groups could significantly increase liver glycogen and muscle glycogen content（P<0.01 or P<0.001）.Compared with the blank group,the activity of Ca²⁺-Mg²⁺-ATPase in the quadriceps of the model group was significantly decreased（P<0.001）.Compared with the model group,the small and medium doses of Kangai injection were all Ca²⁺-Mg²⁺-ATPase activity had no significant effect（P>0.05）,and the high dose group could significantly increase the Ca²⁺-Mg²⁺-ATPase activity in mice（P<0.01）.Compared with the blank group,the expressions of MAFbx and MuRF1 mRNA in the gastrocnemius of the model group were significantly increased（P<0.01）.Compared with the model group,the small,medium and large doses of Kangai injection could significantly inhibit the MAFbx of the gastrocnemius muscle.MuRF1 mRNA expression（P<0.05 or P<0.01）.Results:Protective effect and mechanism of Kangai injection on depression in tumor-bearing miceCompared with the blank group,the number of spontaneous activities,tail suspension time,forced swimming latency and forced swimming time of the model group were significantly lower（P<0.01）.Compared with the model group,Kangai injection low dose group was independent of the mice.The number of activities,the duration of suspending tail,forced swimming latency and forced swimming time had no significant effect（P>0.05）.The medium and large dose groups and the positive drug group could significantly increase the number of spontaneous activities of mice,prolong the time of suspending tail and force Swimming incubation period and forced swimming time（P<0.05 or P<0.01）.On the 1st,7th,14th and 21st day of the experiment,the average weight of the mice in the blank group increased;the body weight of the model group,Kangai injection small,medium and large dose groups and the positive drug group decreased to varying degrees.Compared with the blank group,the hippocampus weight of the model group was significantly lower（P<0.05）.Compared with the model group,the small and medium doses of Kangai injection had no significant effect on the hippocampus weight of the mice（P>0.05）.The positive drug group can significantly increase the hippocampus weight of mice（P<0.05）.Compared with the blank group,the 5-HT and NE contents in the hippocampus of the model group were significantly lower（P<0.05 or P<0.001）.Compared with the model group,Kangai injection each dose group,positive drug group mice hippocampus 5 The contents of HT and DA were significantly increased（P<0.05 or P<0.01）,and had no significant effect on GABA content（P>0.05）.Compared with the blank group,the AchE activity of the model group was significantly increased（P<0.01）.Compared with the model group,the activity of AchE in the hippocampus of Kangai injection high-dose group and positive drug group was significantly decreased（P<0.05 or P<0.01）.Compared with the blank group,the serum levels of IL-6 and IL-1βin the model group were significantly increased（P<0.05 or P<0.01）.Compared with the model group,Kangai injection small,medium and large dose groups were compared.IL-6,IL-1βcontent can be decreased（P<0.05 or P<0.01）.Compared with the blank group,the expression of IL-6 mRNA and IL-1βmRNA in the hippocampus of the model group were significantly increased（P<0.01）.Compared with the model group,Kangai injection can be used in small,medium and large dose groups.The expression of IL-6 mRNA and IL-1βmRNA was decreased（P<0.05 or P<0.01）.Compared with the blank group,the protein expression of Bcl-2,BAX,caspase-3 and caspase-9 in the hippocampus of the model group increased.Compared with the model group,the small,medium and large doses of Kangai injection could make Bcl-2.The expression of BAX,caspase-3 and caspase-9protein was decreased.HE staining showed that the cells in the blank group were arranged neatly and tightly,and the nucleolus was clear.Occasionally,the individual nuclei were concentrated and there was no obvious neuronal damage.The hippocampal neurons in the model group showed obvious degeneration and necrosis neurons and edema,and the cytoplasm showed uniform eosinophilic staining.The nucleus pyknosis,the intercellular space increased;the Kangai injection each dose group and the positive drug group occasionally seen neuronal eosinophilic lesions,visible scattered denatured neurons,showing a slight shrinkage of the cell body.conclusion:1.Kangai injection can significantly prolong the running time,running distance,rotating time,climbing time of tumor-bearing mice,improve the ability of lactic acid scavenging,increase the activity of Ca²⁺-Mg²⁺-ATPase,and increase hepatic glycogen and muscle glycogen.The reserve,which reduces the expression of protein atrophy genes and enhances the ability of anti-oxidative stress,suggests that Kangai injection can improve the exercise capacity of tumor-bearing mice,enhance the energy reserve of the body,and has a good effect of alleviating cancer-induced fatigue.2.Kangai injection can prolong the forced swimming time,tailing time and the number of spontaneous activities of tumor-bearing mice,inhibit the atrophy of hippocampus in mice,increase the content of monoamine transmitters,and inhibit the expression of IL-6 and IL-1βin hippocampus.Expression inhibits neuronal degeneration in the hippocampus.It is suggested that Kangai injection can alleviate the cancer-induced depression in mice,which may be related to inhibiting apoptosis of hippocampus,reducing inflammatory factors and inhibiting hippocampal neuron degeneration.