| ObjectiveIn this study,a variety of statistical methods were used to analyze the cases of senile non-severe aplastic anemia treated mainly by Danggui Buxue decoction from the aspects of syndrome type,symptom,dosage of astragalus membranaceus and Angelica sinensis,drug use frequency,drug clustering,syndrome differentiation and symptomatic drug use.To systematically summarize the experience of Danggui Buxue decoction in the treatment of senile non-severe aplastic anemia.From the point of view of mitochondrial autophagy,the protective mechanism of Danggui Buxue decoction on bone marrow hematopoiesis in mice with non-severe aplastic anemia was discussed,which provided a new idea and method for the treatment of non-severe aplastic anemia.Method1.Clinical study:109 cases of senile non-severe aplastic anemia treated mainly by Danggui Buxue decoction were input into the traditional Chinese medicine case analysis system,and the statistical analysis was carried out by using the traditional Chinese medicine case analysis system,SPSS and SAS statistical software.The analysis contents include:(1)the dose statistics of Astragalus membranaceus and Angelica sinensis in Danggui Buxue decoction,as well as the frequency statistics and cluster analysis of drugs,taste,drugs and meridians;(2)syndrome type,syndrome element statistics and main syndrome types,syndrome elements and Angelica sinensis tonifying blood.The correlation analysis of decoction;(3)the statistics of the main symptoms and the correlation analysis between the main symptoms and the addition of Danggui Buxue decoction.2.Experimental study:105 healthy male ICR mice were randomly divided into normal group,model group,low dose group,middle dose group and high dose group,with 21 mice in each group.In addition to the normal group,the other groups of mice were treated with cyclophosphamide and benzene for 30days to establish non-severe aplastic anemia mouse model.Six mice in each group were killed immediately(D30)after verification of the model for detection.The remaining mice were treated with Danggui Buxue decoction(2.7g·kg-1·d-1,5.4g·kg-1·d-1,10.8g·kg-1·d-1)in the low dose group,middle dose group and high dose group,respectively,and the mice in the low dose group,middle dose group and high dose group were treated with Danggui Buxue decoction(Danggui Buxue decoction).Normal group and model group were given equal volume 0.11ml·10g-1·d-1normal saline intragastrically,and each group was treated continuously for 14 days.The next day(D45)was set as the sampling time.To observe the effects of different doses of Danggui Buxue decoction on general symptoms,peripheral blood leukocytes,hemoglobin,platelets and the number of bone marrow mononuclear cells in unilateral femur of non-severe aplastic anemia mice,and to observe the effect of Danggui Buxue decoction on bone marrow pathology of non-severe aplastic anemia mice under optical microscope.The effect of hematopoietic tissue proliferation on the number of mitochondrial autophagy vesicles in bone marrow mononuclear cells of non-severe aplastic anemia mice was observed under transmission electron microscope,and the expression of autophagy associated proteins LC3-II,p62and mitochondrial autophagy associated protein parkin was detected by;Western Blot method.Result1.The results of clinical study on 109 elderly cases treated mainly with Danggui Buxue decoction showed that:The main results were as follows:(1)the average dosage of Astragalus membranaceus was(30.48±13.40g)g,the number of Angelica sinensis was 30g,the average dosage of Angelica sinensis was(12.33±4.68)g,the average number of Angelica sinensis was 15g,the average value of Astragalus membranaceus:Angelica sinensis was(2.771.60):1,the number of Angelica sinensis was 3:1.(2)A total of 207 kinds of traditional Chinese medicine were used in 109cases of senile patients treated mainly with Danggui Buxue decoction,among which warm medicine,sweet medicine,spleen meridian medicine and deficiency tonifying(invigorating qi,blood,yang and yin)were the most.(3)there were 40 kinds of drugs used more than 10 times except Astragalus membranaceus and Angelica sinensis,which could be divided into 12 categories by varclus cluster analysis,and their classification accorded with clinical practice.(4)according to the statistics of syndrome elements,30 kinds of TCM syndromes and 12 kinds of syndrome elements were obtained.Combined with clinical practice,logistic regression analysis was carried out between kidney yang deficiency syndrome,kidney yin deficiency syndrome,qi and blood deficiency syndrome,essence deficiency,fire toxin and blood stasis syndrome and 40 kinds of high frequency drugs added.It is concluded that the respective logistic regression equations are in line with the clinical practice.(5)according to the statistics of the main symptoms,there were 54 kinds of symptoms except tongue coating pulse.logistic regression analysis was carried out between the eight main symptoms of muscle bleeding,anorexia,soreness of waist and knee,cold and cold,edema,loose stool,sleeplessness and fever,and 40 kinds of high frequency drugs.It is concluded that the respective logistic regression equations are in line with the clinical practice.(6)the light tongue is the most common tongue,and the weak pulse,string pulse and heavy pulse are the most common pulse images.2.Experimental results.The main results were as follows:(1)low,middle and high doses of Danggui Buxue decoction could improve the general symptoms of non-severe aplastic anemia mice.(2)low,middle and high doses of Danggui Buxue decoction could increase the number of white blood cells,hemoglobin,platelets and bone marrow mononuclear cells in peripheral blood of non-severe aplastic anemia mice to varying degrees(p<0.01).(3)the hematopoietic tissue structure of bone marrow in the non-severe aplastic anemia model group was significantly lower than that in the normal group(p<0.01),and the degree of hematopoietic tissue proliferation in the non-severe aplastic anemia model group was significantly lower than that in the normal group(p<0.01).The hematopoietic tissue structure of bone marrow in the low,middle and high dose groups of Danggui Buxue decoction was recovered to some extent compared with the model group.The degree of hematopoietic tissue proliferation increased in turn p<0.01 compared with the model group,and p<0.01 between the low,middle and high groups).(4)the number of mitochondrial autophagy vesicles in the model group was significantly lower than that in the normal group(p<0.01).Compared with the model group,the mitochondrial autophagy vesicles in the low,middle and high dose groups increased in turn,and the difference was statistically significant(p<0.05).There was significant difference among the low,middle and high dose groups(p<0.05).(5)compared with the normal group,the expression of LC3-II protein in bone marrow mononuclear cells of the model group was significantly decreased and the expression of p62 protein was significantly increased by western blot(p<0.05),and the expression of p62 protein in bone marrow mononuclear cells of the model group was significantly higher than that of the normal group(p<0.01).Compared with the model group,Danggui Buxue decoction could up-regulate the expression of LC3-II protein in both middle and high groups.The expression of p62 protein was down-regulated(p<0.05),there was no significant difference between the low dose group and the model group(p>0.05),but there was no significant difference between the middle and high dose groups(p>0.05).(6)compared with the normal group,western blot showed that,The expression of parkin protein in bone marrow mononuclear cells of model group was significantly decreased(p<0.01),and the expression of parkin protein was significantly up-regulated in low,middle and high groups of Danggui Buxue decoction(p<0.05),and the expression of Danggui Buxue decoction was significantly up-regulated in low,middle and high groups.The effect increased with the increase of dose(p<0.05).ConclusionThe main results are as follows:(1)in this study,frequency statistics,varclus cluster analysis,logistic regression analysis and other data mining methods were used to analyze the cases of senile non-severe aplastic anemia treated mainly by Danggui Buxue decoction.This paper makes a systematic summary from the aspects of syndrome type,symptom,dosage,frequency,clustering,syndrome differentiation and symptomatic medication of Astragalus membranaceus and Angelica sinensis,which can be used to guide clinical practice.(2)one of the theoretical mechanisms of non-severe aplastic anemia lies in the existence of mitochondrial autophagy deficiency,which may be mediated by parkin.Danggui Buxue decoction can regulate and restore the bone marrow hematopoietic function of non-severe aplastic anemia model mice by regulating parkin-mediated mitochondrial autophagy,and the effect is positively correlated with the dose. |
PDF Full Text Request