| Objective Hepatic encephalopathy (HE) of type A is a kind of encephalopathies associated with acute liver function failure. It is dangerous and the survival rate of it is only 20%. At present, ammonia intoxication is still in the center of the pathogeneses of HE. Usually, it was thought that ammonia caused HE by interacting brain energy. Recently, the domestic and abroad scholars think the main pathogenesis of HE maybe the disorders of astroglia structure and functions caused by ammonia. In Traditional Chinese Medicine (TCM), it was called "GanJue" . Based on TCM theories and clinical experiences, my tutor -professor Zhang ChiZhi think that the pathogenesis of Ganjue is the result of heat and phlegm troubling mind which caused by the disorders of liver, speel and intestine. The sick position of it is liver, heart, brain and stomach. According to TCM therapeutic principle of "liver and large intestine are collected with each other, diseases of liver are fit to clean large intestine " and "be ill upper, treat down" , professor Zhang uses wined Rhubarb and Lactohacillin to make RuHuang Pill. Clinic and animal studies showed it had good effects on the treatment of HE. It could decline ammonia concentration, obviously ameliorative the symptoms and signs of HE, increase survive rate. The study is to observe the effects of RuHuang pill on the structure and protein expressions of astroglia, to reveal the function point and mechanism of it. Methods(l)96 male Wistar rats weighted 200±20g of SPF grade were randomly divided into six groups: Normal control group (group A), model group (group B), lactulose group (group C), RuHuang Pill group of small dose (group D), RuHuang Pill group of middle dose (group E), RuHuang Pill group of large dose (group F). After fasted overnight for over 14h, all rats except group A were induced to be model of HE of type A by administration with TAA 250mg/ kg (twice, per 24 h once). Before two hours of administration with TAA, each rat in group A and group B were given normal saline 1ml/100g weight, and the left were given the same volume/weight medicine respectively, then redone every 8h. From the first administration with TAA to the end of experiment, every rat was injected mixture solution contained 10% glucose 3ml, saline 1ml, and 20umol/l KC1 lml every 8h to avoid hypoglucose. At 30h of the experiment, electroencephalogram (EEG) examination was done in each group, and then all rats were sacrificed. During the experiment, the behaviors of all rats were observed closely. (2) Blood from eye socket was used to examine the concentrations of serum ALT and blood ammonia. (3) The permeability of blood-brain barrier (BBB) was detected by Evans Blue (EB) leakage method, and the ultrastruncture of BBB, which contained astroglia, was observed under electron microscopy. (4) The cerebrospinal fluid collected from cerebellommedullary cistern was used to determine the concentrations ofammonia. Cerebral cortexes homogenized were used to examine the contents of glutamine by high performance liquid chromatography. Whole brain tissues were used to determine the percentages of brain water content by dry-wet method. (5)The expressions of glial fibrillary acidic protein (GFAP) in cerebral cortex, which was an important structure protein of astroglia, were observed via immunocytochemistry. (6)The expressions of GLT-1mRNA in cerebral cortex, which was a kind of astroglia glutamate transporters, were detected by reverse transcription-coupled polymerase chain reaction (RT-PCR). (7) Cerebrospinal fluid of RuHuang Pill was prepared to effect astroglia CRL-2541 exposed to 5mM concentration of NH4C1. After 48h treatment, the MTT tests were performed to observe the effect of drug on the rate of CRL-2541 survival, immunocytochemistries were performed to examine the expressions of GFAP. Result (1) RuHuang pill could obviously ameliorative the disorders of behaviors and EEGs of rats with HE of type A, decline the concentrations of serum ALT and blood ammonia(P<0.01), increase survive rates. Three dose groups of RuHuang P... |
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