| Objective:In recent years,the incidence of malignant tumors has increased yearly and has become a common disease threatening human health.Although chemotherapy is a major means of malignant tumor therapy,the incidence rate of thrombocytopenia caused by chemotherapy is high,which has a restrictive effect on the continued treatment and impairs the quality of life of patients suffering from malignant tumors.At present,there are few kinds of drugs approved for the treatment of chemotherapy-induced thrombocytopenia.In addition,biological drugs,chemical drugs and proprietary Chinese medicines have their own shortcomings.The efficacy of platelet transfusion with biological drugs is stable,but there are many adverse reactions,and the price is high.Among the chemical drugs,rh IL-11 and rh TPO are approved by the State Drug Administration for marketing,but only rh IL-11 which is also approved by the FDA of the United States for the treatment of CIT can be used for the treatment of CIT.Therefore,rh IL-11 is widely used in clinic,and its action intensity and onset time are poor.There is no Chinese patent medicine specifically for the pathogenesis of"spleen and kidney yang deficiency,blood stasis and blood deficiency"in patients with thrombocytopenia caused by chemotherapy for malignant tumors.Therefore,there is an urgent need for more safe,effective and economical drugs for chemotherapy-induced thrombocytopenia.This study is an effective experience formula for chemotherapy-induced thrombocytopenia that my mentor and his team have gradually sorted out,summarized and screened out in many years of clinical work in preventing and treating malignant tumors with traditional Chinese medicine.Based on the clinically proven safe and effective"Si Xian Prescription",the"Si Xian Granule",a traditional Chinese medicine preparation of medical institutions with the functions of warming the kidney,strengthening the spleen,promoting blood circulation and generating blood for chemotherapy-induced thrombocytopenia,is developed to make up for the deficiency of the current drugs for chemotherapy-induced thrombocytopenia,and to meet the urgent needs of clinical Chinese medicine preparation for the safe,effective and economic treatment of chemotherapy-induced thrombocytopenia.At the same time,it can give play to the advantages of traditional Chinese medicine in improving the toxic and side effects of malignant tumor treatment,supporting chemotherapy treatment,and improving the quality of life of patients,so as to meet clinical needs.In this study,the mechanism of SI XIAN Prescription’s function of"warming the kidney and strengthening the spleen,promoting blood circulation and generating blood"on chemotherapy-induced thrombocytopenia was expounded through animal experiments from peripheral blood to the hematopoietic system.Methods:According to the efficacy of the drug,SI XIAN Prescription was disassembled.The kidney warming and spleen strengthening group was composed of Curculiginis Rhizoma,Epimedii Folium,Ganoderma,Poria,Astragali Radix,and Jujubae Fructus.The blood activating and blood generating formula was composed of Agrimoniae Herba,Spatholobi Caulis,Curcumae Radix,and Forsythiae Fructus,and was extracted into 0.4g crude drug/m L solution.A total of 120 SPF KM mice were randomly divided into 6groups:normal control group,model group,rh IL-11 group(250μg/kg),Si Xian Prescription group(20 g/kg),the kidney warming and spleen strengthening group(KWSSG,12 g/kg)and the group of promoting blood circulation and generating blood(GPBCGB,8 g/kg).From the first day of the experiment,the normal control group and the model group were given distilled water by gavage;the Si Xian Prescription group,the KWSSG and GPBCGB were given corresponding drugs by gavage,with the gavage volume of 20 m L/kg;Rh IL-11group was subcutaneously injected with recombinant human interleukin-11solution,and the injection volume was 10 m L/kg.The drug was continuously administered for 14 consecutive days,once a day.On the 8th day,except the normal control group,the mice in other groups were injected with carboplat in solution in the tail vein with a concentration of 5 mg/m L and an injection volume of 0.15 m L/10g for the purpose of establishing a chemotherapy-induced thrombocytopenia mouse model.The mice in the normal control group were injected with normal saline through the tail vein according to the same administration volume.On the 14th day and 40 minutes after the last administration,the blood of the mouse orbital vein was collected,and the number of platelets was measured by the animal blood cell analyzer.The mice were killed after cervical spondylosis,and the right femur was separated.The bone marrow cavity was repeatedly washed with 1m L RPMI 1640 culture solution with No.6 needle.After washing and staining,the proportion of CD34~+and CD41~+CD61~+positive cells was detected by BDC6 flow cytometry.The left femur was separated and fixed in 10%formaldehyde for 24 h.After decalcification,paraffin was embedded and cut into 5μm thick sections.Then the cells were stained with hematoxylin and eosin(H&E)and photographed under a microscope to analyze megakaryocyte numbers.Results:The results of the peripheral blood platelet count test of mice with chemotherapy-induced thrombocytopenia by Si Xian Prescription and its disassembled formula were statistically analyzed by SPSS22 software.Compared with the normal control group,the number of platelets in the model group was significantly decreased(p<0.01),indicating that the model was successful.Compared with the model group,the number of platelets in each administration group was higher than that in the model group(p<0.01,p<0.05),and the number of platelets in Si Xian Prescription group was(1048±230)×109/L the highest.The expression rate of CD34~+cells in bone marrow of mice with chemotherapy-induced thrombocytopenia treated with Si Xian Prescription and its disassembled formula showed that the proportion of CD34~+cells in bone marrow cells of mice in the model group was significantly decreased compared with that in the normal control group(p<0.01).Compared with the model group,the proportion of CD34~+cells in each treatment group was higher than that in the model group(p<0.01,p<0.05).The expression rate of CD34~+in bone marrow cells of mice in Si Xian Prescription group(0.35±0.17)%was the most significant.The expression rate of CD41~+CD61~+in bone marrow cells of mice with chemotherapy-induced thrombocytopenia treated by Si Xian Prescription and its disassembled formula showed that the proportion of CD41~+CD61~+cells in bone marrow cells of mice in the model group was significantly lower than that in the normal control group(p<0.01).Compared with the model group,the number of platelets in each administration group was higher than that in the model group,and the data of Si Xian Prescription group was of statistical significance.The detection of the megakaryocyte number in the mice femur with chemotherapy-induced thrombocytopenia treated by Si Xian Prescription and its removal showed a significant reduction in the number of megakaryocytes in the normal control group(p<0.01);the number of platelets in each treatment group was increased compared with the model group(p<0.01,p<0.05).Conclusion:1.Carboplatin injection was used to model KM mice,and the test results were statistically analyzed.Compared with the normal control group,the number of platelets in the model group was significantly reduced(p<0.01),indicating that the model was successful and could be used to study chemotherapy-induced thrombocytopenia.2.The Si Xian Prescription group achieved good results on the peripheral blood cell count of chemotherapy-induced thrombocytopenia mice,the expression rate of mouse bone marrow CD34~+cells,the expression rate of mouse bone marrow CD41~+CD61~+cells and the detection of the megakaryocyte number in the mice femur.3.Equivalent to the data in the group of rh IL-11,the data in the group of warming the kidney and strengthening the spleen are slightly lower than those in the group of Si Xian Prescription,indicating that Si Xian Prescription can still be improved.On the other hand,it shows that the"warming the kidney and strengthening the spleen"method is an important principle for the treatment of chemotherapy-induced thrombocytopenia.To sum up,with the function of"warming the kidney and invigorating the spleen,promoting blood circulation and generating blood",Si Xian Prescription has a desirable curative effect on chemotherapy-induced thrombocytopenia.Its formulation is scientific and reasonable,which is worthy of being widely used in clinical practice. |
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