Based On The "kidney-brain" Related Theory, The Mechanism Of Action Of Liuwei Dihuang Decoction On HIBD In Neonatal Rats Was Studied

Research BackgroundHypoxic-ischemic encephalopathy (HIE) refers to hypoxic-ischemic brain injury (HIBD) caused by perinatal asphyxia. It expresses a series of clinical CNS abnormalities [1], such as disturbance of consciousness, convulsions, muscle tension changes and others, which often left varying degrees of neurological sequelae after treatment. At present, domestic and foreign HIE pathogenesis is still not completely clear. Energy metabolism, oxygen free radicals, calcium overload, the toxic effects of excitatory amino acids, inflammation, apoptosis and other mechanisms are studied more at present. On the basic of "three support" and "three symptomatic", the research on emerging drugs and therapies such as neurotrophic factor, free radical scavenger, erythropoietin, and stem cell transplantation therapy are favored by people,in addition to the conventional high-pressure oxygen, hypothermia, brain cell metabolism activating agent therapy and drugs. But they mostly limited to animal experiments, which not widely used in clinical practice.Increasing numbers of report in Chinese medicine treatment of HIE are reported. And they made some effect, such as Salvia injection, TMP, which are widely reported and proved their clinical efficacy. Yet they are growing into a "Chinese medicine with Western" position, which large numbers of reported drug treatment in HIE is the use of blood circulation that lacks the differential treatmentin traditional Chinese medicine philosophy. In addition, they limit the location of HIE only in the brain, which lacks overall concept of Chinese medicine. But"kidney - brain" related in Chinese medicine both riches theoretical support and wide ranges of clinical practice. Therefore, this study attempts to use Liu Wei Di Huang Tang which is the representative prescription of tonifying kidney for the treatment of HIEin encephalopathyin western medicine.Research purposeObserving therapeutic effect of Liu Wei Di Huang Tang in treating newborn rats HIBD and explore its mechanism.While verifying the theory of TCM "kidney - brain" is related and upper part diseases can be treated with lower locations, it also provides new ideas and modalitiesfor the TCM treatment of encephalopathyin modern medicine.literature ResearchLiterature cited similar pediatric disease name with HIE, such as fetal weak, dream life, etc., and elaborated its pathogenesis and treatment principles. So it supposes there are two main reasons for the occurrence of these diseases:Fetal internal factors and exogenous factors. But internal factors are the nature, external factors are standard. So the best means of treatment of fetal diseases are found the kidney, fill lean marrow and conditioning the internal. On this basis, we fully demonstrates the "kidney-brain" associated with rationality from five areas which are the historical origins, the material base, the meridians affiliate, clinical research and experimental research. Finally, the modern pharmacological research as well as its progress in the treatment of encephalopathy modern Liuweidihuang soup was also discussed.Experiment ResearchExperiment 1:The Health Level 7-day-old SPF SD rats 88, male or female, weighing 13-18 g were randomly divided into three groups:HIBD model group (32), sham operation group (32), Chinese intervention group (24). The first two groups depending on the time of death are divided into four subgroups:2h, 1d,3d, and 7 d, which n= 8 in each group.Chinese medicine group was divided into three subgroups:1 d,3d, and 7d. The model group and Chinese medicine group were used classical methods of Rice-Vannuccimodeling. After modeling, the behavior of the ability to change the situation was observed. The Chinese medicine group was given 0.3 ml/10g volume Chinese medicineorally once daily which was continuously being fed 7d. The model group and the sham group were given same dose of normal saline. Each group were removed its brains at different time points, which were used HE staining in paraffin section to observe pathological changes in brain tissue. In experiment Ⅰ and Ⅲ, the Grouping, modeling, gavage method, and the average time putting to death are the same as experimental I. In experiment Ⅱ, femoral artery blood serum was separatedbefore decapitated. After that, samples malondialdehyde, superoxide dismutase, and neuron-specific enolase(MDA, SOD, NSE) levels were measured at different time pointswith enzyme-linked immunosorbent assay (Elisa). In experimental III, brain tissue gene expression of HIF-1α was measured by reverse transcription polymerase chain reaction (RT-PCR).Results1. The ability to change the behavior: There are great changes in capacityin and after the modeling process in model group, especially performed with the contralateral limb movement disorder, clip tail dextrose, post ptosis, and difficulty in opening eyes and other changes in characteristics. Sham group has no similar situation. Drug group is similar to the model group in the first 3 days, but is similar with the sham groupthe late performance.2. HE staining:in model group, the number ofpyramidal cellsin hippocampal Lesion sidedecreased, array disordered, nuclei dissolved and disappear,cell body decreased, some cells in hippocampus has visible edema and vacuolar degeneration. In sham group, there are very slight pathological changes also, butthey gradually repaired. In drug group,pathological changes are the same as model groupin the beginning, but they gradually restored later, which close to sham group.3. NSE:In model group, serum NSE levels increased significantly at 2h, and gradually increased over time. Insham group,there was no change significantly.In Chinese medicine group, it decreasedgradually. when Chinese medicine groupcompared with the model group, the difference was statistically significant in 3d and 7d (p<0.05, p<0.01).4. MDA: MDA levels had no significantly difference in 2h in model group and sham group. Compared medicine group to model group.there was a significant change in 1d (p<0.01), while no significant changewhen compared tothe sham group, but medicine group and the model group had no significant difference in each latertime point.5. SOD:There was no significant change in SOD levelsin 2h. However, Comparedthe model group withChinese medicine group, it hada significant difference, and SODlevels in Chinese medicine groupwas close tothe sham group in 1d,3d, and 7d (p<0.01).6. HIF-la:HIF-la gene expression increasedat 2hin model group, decreased at 1d, and gradually rose after that. Compared with the traditional Chinese medicine group,the model grouphad a significant difference in 3d and 7d (p<0.05). HIF-1α gene expression levels in Chinese medicine groupis closer to the sham group.Analysis conclusion1. "kidney - brain" being related and tonifyingkidney to cure brain has a rich theoretical supportand extensive clinical practice, thisprovides a theoretical support using Liu Wei Di Huang Tang to treat HIBDas a treatment.2. Thisstudy successfully completedthe HIBD model of neonatal rats with the method. Its pathological changesis consistent with HIE brain damage.3. Liu Wei Di Huang Tang can promote the repair of lesions in the brain, reducing the concentration of serum NSE, improve SOD activity in brain tissue,and down-regulated the expression of HIF-1α gene. But there’s no significant effect on the content of MDA.4. Liu Wei Di Huang Tang has therapeutic effect on neonatal rats HIBDinjury. The mechanism is related to HIF-1α: by increasing the activity of SOD and reducing MDA level,thus decreasing the expression of HIF-1αto protect hypoxic-ischemic brain tissue, promote repair of damaged neurons, and decrease serum NSE concentrations.