| Objective: This topic in moxibustion clinical curative effect on bone marrow suppression after chemotherapy,on the basis of exact to suppress a tumor-burdened bone marrow in mice as the research object,from the closely related to tumor onset of Wnt signaling pathways of moxibustion on tumor mice bone marrow suppression after chemotherapy Wnt signaling pathways key protein Wnt5a,beta-catenin expression,the effect of moxibustion on a tumor-burdened mice after chemotherapy the change of the blood cells,bone marrow suppression for moxibustion intervention provide biological mechanisms,conversion treatment strategies for bone marrow suppression after chemotherapy patients.Method: SPF KM mice,50 males,weighing 15-20 g.Ten blank groups were set,and all the other mice were inoculated with tumor strains.After successful modeling,the mice were divided into tumor bearing model group,cyclophosphamide group,shalliver alcohol group and moxibustion treatment group.Except for the tumor-bearing model group,all the other groups were given cyclophosphamide(CTX)100mg/kg/d for intraperitoneal injection to establish the bone marrow suppression model in tumor-bearing mice.After successful modeling,the shark liver alcohol group was treated with intraperitoneal injection of shark liver alcohol solution,and the moxibustion group was treated with moxibustion Zusanli,3min per point each time,once a day,for a total of 10 days.After the treatment,blood was collected from the eyes of mice,and the number of red blood cells,white blood cells and platelets was detected by automatic blood detector.The tumor tissue was stripped,the tumor weight was weighed,and the tumor inhibition rate was calculated.The thymus gland and spleen were stripped to calculate the organ index of mice.One side of femur of mice was taken,and the effects of moxibustion on bone marrow tissue of tumor-bearing mice after chemotherapy were observed by HE staining and immunohistochemistry.Wnt5a and β-catenin proteins were used as detection indexes to explore the effects of moxibustion on Wnt signaling pathway.Results:1 General observation of mice in each groupIn the blank group,there was no modeling intervention in the whole experiment,and the mice in all conditions,including eating and drinking water,were in a healthy state.They were flexible and responsive without any abnormal conditions,and their body size was slightly larger than that of the other groups.The tumor-bearing model group was inhibited.In the experiment,except for some movement problems caused by the growth of armpit tumors,there was no hair loss,and the mental state was good,and the drinking and eating were normal.One mouse died on the 8th day of intervention treatment.Mice in CTX group had reduced food and water consumption,weight loss,low energy,lying in piles with fatigue,and dull fur.Three days after the establishment of the cyclophosphamide model,hair loss began to appear,with slow activity,arched back,pale eyelids,and occasional blood in stool.One mouse died on the 6th and 9th day of treatment respectively.The shark liver alcohol group: Compared with the cyclophosphamide group,the spirits of mice in the shark liver alcohol group were improved,and the eating and drinking conditions were improved.Some mice showed symptoms of eyelid inflammation and slight irritability.One mouse died on the 7th and 9th day of treatment respectively.The moxibustion group: compared with the cyclophosphamide group,the above conditions of mice in the moxibustion group recovered to a certain extent,and their spirits improved.A few mice still had irritability,fatigue and lying in piles,arching their backs,eating and drinking water gradually normal,and their movements were flexible.2 Changes in the number of peripheral blood cells in miceCompared with the blank group,there was no significant change in peripheral white blood cells of the tumor-bearing model group(P>0.05),the difference was not statistically significant;lymphocytes were significantly decreased(P<0.01),the difference was statistically significant;platelets were significantly increased(P<0.01).The white blood cells and lymphocytes of CTX group were significantly decreased(P<0.01),and the difference was statistically significant(P>0.05).Compared with the blank group,there were no significant differences in the peripheral blood white blood cells and platelets(P>0.05),and the lymphocytes decreased(P<0.01),the difference was statistically significant.There were no significant differences in the white blood cells and lymphocytes of mice in the moxibustion group compared with the blank group(P>0.05),and the increase of platelets was significant(P<0.05),the difference was statistically significant.There was no significant difference in the blood white blood cells,platelets and lymphocytes between the shark liver alcohol group and the CTX group(P>0.05).Compared with CTX group,the number of white blood cells in moxibustion group was significantly increased(P<0.05),and the difference was statistically significant,while the difference of platelets and lymphocytes was not statistically significant(P>0.05).3 Comparison of viscera index of mice(1)Tumor weight: Tumor weight in the cyclophosphamide group and the shalanol group was significantly lower than that in the tumour bearing model(P<0.05),and the difference was statistically significant in the moxibustion group(P<0.01).The tumor weight ratio in CTX group was significantly increased(P<0.05),and the difference was statistically significant.(2)Organ index: spleen weight,spleen index and thymus weight of mice in the tumor-bearing model group were increased compared with those in the blank group(P<0.01),and the thymus index of mice in the tumor-bearing model group was significantly higher than that in the blank group(P<0.05),the difference was statistically significant.The spleen weight,spleen index and thymus weight of CTX group were significantly increased compared with blank group(P<0.01),and the difference was statistically significant.The spleen weight,spleen index and thymus weight of mice in shiganitol group were significantly increased compared with blank group(P<0.01),and the difference was statistically significant.The spleen weight and spleen index of mice in moxibustion group were significantly increased compared with blank group(P<0.01),and the difference was statistically significant.There was no significant difference in the organ index between the squagliol group and the CTX group(P>0.05).The spleen weight and spleen index of mice in moxibustion group were significantly lower than those in CTX group,and the difference was statistically significant(P<0.05).4 Morphological changes of bone marrow in mice(1)There were a large number of granulocytes in the bone marrow hematopoietic tissue of mice in blank group,with complete structure and clear,uniform and compact arrangement of megakaryocytes;(2)Compared with the blank group,there were a large number of inflammatory cells in the bone marrow hematopoietic tissue of mice in the model group,and the hematopoietic area was reduced.(3)Compared with the model group,there were fewer inflammatory cells in the bone marrow cells of mice in the cyclophosphamide group,but compared with the blank group,the hematopoietic area in the cyclophosphamide group was significantly reduced,the morphology of bone marrow cells in the bone marrow tissue of mice was abnormal,and the blood sinuses were damaged.(4)Compared with the cyclophosphamide group,the morphology of bone marrow cells in the shalliol group and the moxibustion group was generally normal,the hematopoietic area was significantly increased,and the inflammatory cells were also significantly reduced.5 Expression of Wnt5a and β-catenin in mouse bone marrow cells(1)Wnt5a: Compared with the blank group,the expression of Wnt5a protein in bone marrow cells of the tumor-bearing model group was increased(P<0.01),and the difference was statistically significant.The expression of Wnt5a protein in CTX group was higher than that in blank group(P<0.01),and the difference was statistically significant.Compared with the blank group,the expression of Wnt5a protein in bone marrow cells of mice in the squagliol group was decreased(P<0.01),and the difference was statistically significant.Compared with CTX group,the expression of Wnt5a protein in bone marrow cells of mice was significantly decreased(P<0.01).The difference was statistically significant.The expression of Wnt5a protein in bone marrow cells of mice in moxibustion group was lower than that in CTX group(P<0.01),and the difference was statistically significant.(2)β-catenin: The expression of β-catenin protein in bone marrow cells of mice in model group was significantly higher than that in blank group(P<0.01),and the difference was statistically significant.The expression of β-catenin protein in bone marrow cells of mice in CTX group was significantly higher than that in blank group(P<0.01),with statistical significance.Compared with the blank group,the expression of β-catenin protein in bone marrow cells of mice in the squagliol group was significantly decreased(P<0.01),and the difference was statistically significant.The expression of β-catenin protein in bone marrow cells of mice in moxibustion group was significantly lower than that in blank group(P<0.01),and the difference was statistically significant.There was no significant difference in the expression of β-catenin protein in bone marrow cells between the squagliol group and the CTX group(P>0.05).The expression of β-catenin protein in bone marrow cells of mice in moxibustion group was significantly lower than that in CTX group(P<0.01),and the difference was statistically significant.Conclusion:1.Moxibustion Zusanli can significantly improve the level of white blood cells in mice after CTX chemotherapy,improve the mental and dietary status of myelosuppressant mice,and improve the life quality of mice;The effect of moxibustion Zusanli on restoring spleen index and inhibiting tumor rate of mice is better than that of shark liver alcohol.2.Moxibustion Zusanli can significantly improve the damaged state of bone marrow after cyclophosphamide chemotherapy,significantly increase the hematopoietic area of bone marrow and significantly reduce inflammatory cells.3.CTX chemotherapy damages the hematopoietic microenvironment of the bone marrow of mice and promotes the expression of Wnt5a and β-catenin protein in bone marrow cells,which may be one of the mechanisms leading to bone marrow suppression.The repair mechanism of moxibustion Zusanli on bone marrow hematopoietic microenvironment and bone marrow injury in myeloid inhibited mice may be related to the regulation of Wnt signaling pathway key cytokines Wnt5a and β-catenin protein expression. |
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