Ischemic White Matter Damage Jieyu Prescription Intervention Mechanism Of Activation Of Apoptotic Genes And Happy

White matter lesions(WML) is a kind of central nervous system diseases. The major pathological change on brain white matter is that microglial cells are activated. Astrocytes are swellen,glial fibrillary acidic protein (GFAP) are positive,Oligodendrocytes are injured and even apoptosis,Leukoaraiosis(LA) and loss of myelin,which can severely consult in abnormality of emotion and behaviour with the patients. With the increase of the cerebral diseases, many patients are developed Vascular depression(VD) and Vascular Cognitive Impainnent(VCI). The mechanism of WML is still been studied, the oxidative stress and inflammatory injuries play important roles in WML. Traditional Chinese Medicine(TCM) considers that VD occurs is closely connected with the deficiency of brain. Reinforcing the function of kidney to replenish marrow, removing blood, phlegming stasis and enriching brain are the regular therapies. Kai Xin Jie Yu fang based on the long-term clinical observation by professor Huang Shijing is composed of Kai Xin San and Si Ni San. It can treat VD and MCI by benifiting Qi and relieving depression, which provides scientific ground and theoretical basis in clinical applications.The 3 main experiments :1 The first part is the observation of learning and memory and the pathology and morphological in white matter: The cerebral ischemia model in rats was established by 2-vessel-occulsion at 3day, 7day and 21 day. The rats were randomly divided into 3 groups: shamed group(shamed). the cerebral ischemia model group(model) and Chinese herbs treatment group(treatment) at every time, the treatments were fed by Kai Xin Jie Fu Fang( 1.8g/kg/day). The rats were killed at 3day. 7day and 21 day. The learning and memory of rats was observed with the Morris maze, and cerebral blood flow(CBF) in frontal lobe and parietal lobe was assessed. The brain was stained by HE to observe the structure of white matter. The brain white matter beyond frontal lobe was soaked into glutaradehyde to observe the ultrastructure of brain white matter by TEM.2 The second part is the observation of apoptasis of neuron cells and expression of Bcl\BaxmRNA and protein The cerebral ischemia model of rats was established by 2-vessel-oceulsion at 3day. 7day and 21 day. The rats were randomly divided into 3 groups: shamed group(shamed),the cerebral ischemia model group(model) and Chinese herbs treatment group(treatment) at every time, the treatments were fed by Kai Xin Jie Fu Fang(1.8g/kg/day). Terminal deoxynucleotidyl Transferase Biotin-dUTP Nick End Labeling ( TUNEL stainning) was used to observe apoptasis in barin. The expression of Bcl\BaxmRNA and protein were assessed by RT-Pcr and Western blot.3 Reactive oxygen species(OS) and Nuclear transcription factorκB(NF-κB):The cerebral ischemia model in rats was established by 2-vessel-occulsion at 3day. 7day and 21day. The rats were randomly divided into 3 groups:shamed group(shamed). the chronical cerebral ischemia model group(model) and Chinese herbs treatment group(treatment) at every time, the treatments were fed by Kai Xin Jie Fu Fang(1.8g/kg/day). NO,NOS and SOD were detected by kits. HNE was detected by ELISA. Bcl-2 and Bax mRNA were investigated by RT-PCR and the protein expression of Bcl-2 and Bax were assessed by Western blot.Experiments results:1. The effects of Kai Xin Jie Yu Fang on learning and memory ability and pathological change of brain white matter in rats with cerebral ischemiaThe ability of learning and memory of rats were decreased at 3day. 7day and 21day and significant difference was found at 3day(P<0.01). Compared with model, the treatment were increased, and significant difference at 3day(P<0.05); BCF of rats was found less in frontal lobe and parietal lobe in model, significant difference at 21day(P<0.01 or P<0.05). compared with model, treatment was dramatically increased, significant difference at 21 day(P<0.05). In shamed the structure of brain matter is abmoral. The tissue of white matter by HE was loosed, the ultrastructure showed that the glial cells degenerated and even died, demyelination and Leukoaraiosis by transmission electron microsopy in model, the structure were better in treatment than the model.2. The effects of Kai Xin Jie Yu Fang on apoptasis of neurons and expression of Bcl-2mRNA and protein BaxmRNA and protein in rats with cerebral ischemiaIn shamed, few apoptasis were found cortex and white matter. More apoptasis appeared in model and increased with the prolonging of time. Compared with model, treatment were less. In model, the expression of Bcl-2 mRNA and protein decreased (P<0.01 or P<0.05). and with peak at 24day. The treatment was increased and had thestatistical significance(P<0.01 or P<0.05) from the model, meanwhile expression of Bax mRNA and protein enhanced and had statistical significance from treatmetn(P<0.01 or P<0.05 ), With time prolonging, the expression constantly ascended and reached its peak at 24day and decresed after treatment of Kai Xin Jie Yu Fang.3. The effects of Kai Xin Jie Yu Fang on Oxidative Stress and expression of NF-κB in rats with cerebral ischemia.Compared with shamed, content of NO and activities of NOS were increased at 3day. 7day and 21 day. significant difference at 21 day(P<0.01 or P<0.05). in treatment, content of NO and activities of NOS fell down, significant difference at 21day(P<0.05). In model the activities of SOD were reduced and the contents of HNE were increased, ignificant difference at 3 day and 21 day(P<0.01 or P<0.05). after treatment with Kai Xin Jie Yu Fang, the activities of SOD evalated and the contents of HNE fell down, significant difference at 3 day(P<0.05). In model, at 3day. 7day and 21day, expression of NF-κB increased and reached the peak at 21day(P<0.01), after treatment with Kai Xin Jie Yu Fang, expression of NF-κB dropped and showed significant differences(P<0.01).CONCLUSIONBrain white lesion was connected with Vascular Depression. 2-VO is the better model that made the brain white lesion. Kai Xin Jie Yu Fang can treat the Vascular Depression. In order to study the mechanism of the Kai Xin Jie Yu Fang. The experiments was made. The results were following:1 The ability of learning and memory were decreased in rats with ischemia. The cerebral ischemia can activate the expression of BaxmRNA and pretein and inhibit the expression of Bcl-2mRNA and protein, which make the apoptasis of oligodendrocyte and the white matter lesion. The results indicated that the ability of learning and memory was related to the white matter lesion. Kai Xin Jie Yu Fang can improve the ability of learning of memory by protecting the brain white matter. 2 The activity of SOD was decreased, contents of FINE was increased and the activity of NOS and the contents of NO were enhanced in rats with crerbral ischemia, which proved that ROS conducted more, the expression of NF-κB was increased. It showed that ROS including NO can activate the expression of NF-κB. Kai Xin Jie Yu Fang can inhibit the expression of NF-κB by releasing ROS lesion.Conclusion, brain white matter was injured after cerebral ischemia. OS took part in the lesion. Kai Xin Jie Yu Fang can protect the brain white matter by decreasing the ROS,inhibiting the activity of NF-κB. reducing the expression of BaxmRNA and protein and improving the expression of Bcl-2mRNA and protein.