| Hemorrhagic apoplexy is a common disease to middle and old aged. It is harmful tomankind because of its high occurrence and death rate. Recent statistics showed that therewere more researches to ischemic apoplexy but less to hemorrhagic apoplexy. This studywas designed to observe the effects of Chinese medicine of "Jiu Nao Ning" Injection toneuron apoptosis caused by experimental hematoma of rabbits and induced by cerebralischemia reperfusion in vitro. Two experimental models were selected in this research andto porvide the experimental bases for clinical application for in-depth study of the curativemechanism to hemorrhagic apoplexy. This research consists of three parts: 1. Study on pathologic histology changes induced by injecting autogenous clot in thebrain of rabbits and effects of "Jiu Nao Ning" Injection on those changes. In this part, male rabbits, weighted from 2.0 to 2.5 kilograms, were divided randomlyinto six groups, those are: normal control group (control); cerebral hematoma group(model); "Jiu Nao Ning" Injection higher dose group (JNN-H); "Jiu Nao Ning" Injectionmiddle dose group (JNN-M); "Jiu Nao Ning" Injection lower dose group (JNN-L) and"Qing Kai Ling" Injection group; respectively. The model of experimental cerebral hematoma was created by injecting autogenousclot in the brain of rabbits. We observed the influence of "Jiu Nao Ning" Injection on brainindex, content of water, and histo-morphology at the third, seventh and fifteenth days of thecerebral hematoma model. TUNEL and transmission electron microscope (TEM) were usedto determine the changes of apoptosis induced by cerebral hematoma in rabbits. 2. The changes of cyto-metabolism and cytomorphology in cultured neuron inducedby cerebral ischemia reperfusion in vitro and effects of "Jiu Nao Ning" Injection on thosechanges. The cultured neurons were also divided into six groups: control group; model group(ischemia 2 hours, and ischemia 2 hours+reperfusion 4 hours); "Jiu Nao Ning" Injectionhigher dose group (JNN-H); "Jiu Nao Ning" Injection middle dose group (JNN-M); "JiuNao Ning" Injection lower dose group (JNN-L) and "Qing Kai Ling" Injection group;respectively. In this part, the culture of cortex neurons and the simulated ischemia & reperfusionmodel were established. Dissociate cortex cells from early postnatal wistar rats (postnatalhour 24) with DMEM were supplemented by horse serum and N3 medium, andnon-neuronal cell division was halted by cytosine arabinoside. As the result, enoughneurons with extensive and synaptically active network were produced. The model wasestablished by hypoxia and glucose deprivation to simulate ischemia, and then return theneurons to the normal condition to simulate reperfusion. We investigated the activity andtransudation rate of LDH of cultured neurons and morphological, DNA as well as RNAchanges following the damage induced by simulated cerebral ischemia and reperfusion invitro, then studied the influence of "Jiu Nao Ning" Injection with biochemical andfluorescence. Moreover, Annexin V and PI staining were applied to detect the changes ofapoptotic rate at early and late stage of apoptosis process by the flow cytometry techniques. 3. Investigating the molecular mechanism of cultured neuron's apoptosis induced bycerebral ischemia reperfusion in vitro and effects of "Jiu Nao Ning" Injection on thosechanges. In this part, same cell model as mentioned above was introduced. The flow cytometrymethod, immunocytochemical techniques were used to observe the influences of "Jiu NaoNing" Injection on mitochondrial membrane potential, intracellular pH value, and theexpression of bcl-2/bax, Fas/FasL, Caspase-3. Furthermore, the expression of bcl-2/baxmRNA was detected in cultured neurons by RT-PCR method at 2hrs after cerebral ischemiaand 4hrs after reperfusion. The main results displayed were as follows: 1. Effects of "Jiu Nao Ning" Injection on experimental cerebral h...
|
PDF Full Text Request