The Research Of Liu Wei De Huang Tang And Si Jun Zi Tang On GMH-IVH,Based On The Theory Of "Treatment From Kidney" And "Treatment From Spleen"

BackgroudIntracerebral hemorrhage is very common in preterm infants,leading death or severe damage on premature brain,with germinal matrix-intraventricular hemorrhage the most common type.Germinal matrix hemorrhage?GMH?occurs in the subependymal?or periventricular?germinal region of the premature brain,and sometimes develops into intraventricular hemorrhage?IVH?.The symptom of GMH-IVH varies,some patients have no symptom except the abnormality in ultrasound or CT test.Some patients show delirium,low muscle tone,reduced movement,irregular breathing.Severe patients show progressive consciousness disorder,apnea,convulsion,disappearance of pupil reflex.The prognosis of GMH-IVH is poor,with sever hemorrhage often leading to death,while approximately 50%of survivors develop hydrocephalus,cerebral palsy,seizures,hemiplegia,learning disabilities,and so on.The mechanisms and molecular pathways that lead to the injury of GMH-IVH is not clear yet.It is generally believed that the vulnerable structure of germinal matrix is the basis of hemorrhage,the resent researches focus on the blood components and metabolites,microglia and inflammation,lymphocytes and immunity.As for the treatment,given the limited acknowledge of GM-IVH,there are only several methods we can choose to deal with it,while the advanced treatments such as neurotrophic drugs,iron chelators,NSAIDs,and stem cells therapy are still in research.Due to the lack of effective drugs right after hemorrhage,the patients are left with sever neurological damage.As there is no report in Chinese medicine treatment of GMH-IVH,we intent to find the possible solution from the Chinese medicine in this research.Research purposeDue to the lack of effective drugs right after hemorrhage,the patients are left with sever neurological damage.In this research,after analysis of Chinese medicine records from ancient time,we choose Liu Wei De Huang Tang and Si Jun Zi Tang based on the based on the theory of“treatment from kidney”and“treatment from spleen”respectively,observe their effect and analyze the mechanisms on GMH-IVH animal model,in order to find the safe and effective recipe and the Traditional Chinese Medicine solution for GMH-IVH.Literature researchThe name of GMH-IVH is based on anatomy,and there's no report in Chinese medicine treatment of GMH-IVH,but the disease dose not exist only in Western medicine.We search the Chinese medicine records from ancient time,cite similar pediatric diseases with GMH-IVH and elaborate their pathogenesis,clinical manifestation and treatment,and decide the possible treatment direction.Then we analyze the mechanism of GMH-IVH,identify the key point of the disease,combine with the results in Chinese medicine records,choose Liu Wei De Huang Tang and Si Jun Zi Tang as the prescription.At last,we elaborate the clinical use of the two prescriptions in related diseases,to understand the scope of application.Experiment researchThe Health Level 7-day-old SPF SD rat pups 160?brain development comparable to 30–32 week gestation infants?,male or female,weighing 11-15g,were randomly divided into4 groups:GMH-IVH model group?n=40?,Liu Wei De Huang Tang group?n=40?,Si Jun Zi Tang?n=40?and sham-operated group?n=40?.The GMH-IVH model group,Liu Wei De Huang Tang group and Si Jun Zi Tang group were injected intraparenchymal of bacterial collagenase?0.3U?to induce GMH-IVH,then given normal saline,Liu Wei De Huang Tang and Si Jun Zi Tang respectively,once a day for 7d.Sham-operated group was injected intraparenchymal of nothing,then given normal saline,once a day for 7d.In experiment?,we investigated the effect of Liu Wei De Huang Tang and Si Jun Zi Tang on the neurological function and the ventricular volume,short-term neurological tests were performed on the 1^st day and the3^rd day after GMH-IVH,while long-term neurological tests were performed on the 28^th day after GMH-IVH.The ventricular volume was measured on the 28^th day by MRI after GMH-IVH.In experiment?,we use hemoglobin assay to test hemoglobin content of the groups to compare their effect on clearing blood clots at 24h and 72h after GMH-IVH.Immunohistochemistry was used to investigate the number of M2 microglial at 72h after GMH-IVH.In experiment?,ELISA was applied for detecting serum IL-1??IL-1 and TNF-?protein at 24h,3d and 7d after GMH-IVH to compare the effect of Liu Wei De Huang Tang and Si Jun Zi Tang on these pro-inflammatory cytokines.Results1.Neurological function:Both Liu Wei De Huang Tang and Si Jun Zi Tang have no effect on short-term neurological function after GMH-IVH.Liu Wei De Huang Tang ameliorates long-term neurological deficits?P<0.05?,while Si Jun Zi Tang has no effect.2.Ventricular volume:GMH-IVH results the increase in ventricular volume,which is significantly reduced in Liu Wei De Huang Tang group 28d after GMH-IVH?P<0.05?,while Si Jun Zi Tang has no effect.3.Body weight:GMH-IVH significantly slows normal growth28d after hemorrhage.Liu Wei De Huang Tang and Si Jun Zi Tang both reduce the body loss significantly?P<0.05?.4.Hematoma resolution:No group have any effect on the hemoglobin content at 24h after GMH-IVH.Liu Wei De Huang Tang group shows less hemoglobin content at 72h after GMH-IVH?P<0.05?.Si Jun Zi Tang group shows no difference with model group.5.M2 microglia:GMH-IVH induces the activation of microglia and causes the tendency to have more M2 microglia compared with sham-operated group at 72h after hemorrhage.Liu Wei De Huang Tang significantly increases M2 microglia compared with model group?P<0.01?.Si Jun Zi Tang shows no difference with model group.6.IL-1?:GMH induces serum IL-1?secretion after hemorrhage,and no group has any effect on it at 24h after GMH-IVH.At 3d and 7d,the serum level of IL-1?is significantly reduced in Liu Wei De Huang Tang group?P<0.05?.Si Jun Zi Tang shows no difference with model group.7.IL-6:GMH induces serum IL-6 secretion after hemorrhage,and no group has any effect on it at 24h after GMH-IVH.At3d and 7d,the serum level of IL-6 is significantly reduced in Liu Wei De Huang Tang group?P<0.05?.Si Jun Zi Tang shows no difference with model group.8.TNF-?:GMH induces serum TNF-?secretion after hemorrhage,and no group has any effect on it at 24h after GMH-IVH.At 3d and 7d,the serum level of TNF-?is significantly reduced in Liu Wei De Huang Tang group?P<0.05?.Si Jun Zi Tang shows no difference with model group.Conclusions1.Liu Wei De Huang Tang can ameliorate long-term neurological deficits and ventricular dilation caused by GMH-IVH,but has no effect on short-term neurological deficits.2.The mechanism of Liu Wei De Huang Tang on long-term neurological deficits:it can increase the number of M2microglia to promote hematoma clearance and decrease the serum level of IL-1??IL-6 and TNF-?caused by GMH-IVH,thus ameliorates long-term neurological deficits.3.In the long term,Liu Wei De Huang Tang and Si Jun Zi Tang both reduce the body loss caused by GMH-IVH significantly.4.The effectiveness of Liu Wei Di Huang Tang in this research to some extent confirmed that“treatment from kidney”is possible.