The Mechanism Of Nao-qing Microemulsion On Intracerebral Hemorrhage In Rats

Research objective Intracerebral hemorrhage(ICH) refers to the primary non traumatic bleeding in brain parenchyma. There is still no effectiveway for the treatment of Intracerebral hemorrhage, which may reduce disability rate and death rate remarkably. Now the research of traditional Chinese medicine on the treatment of ICH is hard to follow the international level, because the cure of ICH is lack of scientific basis, let alone the mechanism of pharmacological fuction. Therefore, there is a great significance to continue this study.We use microemulsion through a spray nasal administration to explore the mechanisim and characteristicof Chinese medicine Nao-Qing microemulsion. With a nasal and brain channel and microemulsion carrier, this study aim to achieve an effect of brain targeting, which will enhance bioavailability, drug compliance as well as promote the improvement of pharmaceutical preparations, with regarding to make a contribution to Chinese medicine treatment of ICH. Content and Result1. The preparation and quality criterion of Nao-Qing microemulsionIn this research, we discussed and determined the formula and preparation technology of Nao-Qing microemulsion. The Nao-Qing microemulsion was belonged to O / W microemulsion, which was brown clear liquid, p H values between 5.5 and 6.5,particle size was 100 nm or less, and no stratification happened after centrifuge with still transparent clear brown microemulsion. The active ingredients of Panax ginsenosides Rg1 and Rb1 were detected by HPLC, and the methodological study for it. The results have shown that the drug content in the Nao-Qing microemulsion with ginsenosides Rg1 as the standard, was 13.78 mg / ml ~ 14.04 mg / ml. Other projects were in line with the General requirements for regulations on nasal preparations in "Chinese Pharmacopoeia " 2010 edition.2. The impact of Nao-Qing microemulsion on behavior and nerve morphology after Intracerebral hemorrhageIn this research, we injected tail arterial blood of rat into its Caudate nucleus to induce a ICH animal model. Then randomly allocated them into different groups, like sham-operation group, group with normal saline, group with Nao-Qing microemulsion. Finally, observed rats, behavior change of each groups after ICH during different interval and learned the change of cellular morphology with paraffin section of brain tissue, provided the basis for our experiment. Result displayed that not only model group but also drug administration group express an symptom of neurological deficit, such as brain tissue nerve cell edema, degeneration, cell space become larger and inflammatory cell infiltration. However, after drug administration, the symptom of neurological deficit of brain hemorrhage rat can be improved.3. The impact on brain edema after Nao-Qing microemulsion administrationDuring this study, brain water content was determined by dry wet weight method and the expressing quantity of AQP-4, MMP-9 proteins was determined by immunohistochemistry staining technique after Nao-Qing microemulsion administration of intracerebral hemorrhage rats model through a special interval, such as 1 day, 3 days, 5 days, 7 days. The result has shown that the water content peak of intracerebral hemorrhage group was appeared on the third day, the seventh day it declined obviously which still above the normal level. Nao-Qing microemulsion administration group, intracerebral hemorrhage model brain tissue water content in 1d, 3 d, 5 d, 7 d time points higher than the sham group(P <0. 05), the brain clear microemulsion containing treatment group water were lower than at the corresponding time points ICH model group(P <0 05.); expression of brain hemorrhage model group AQP-4 / MMP-9 in 1d, 3 d, 5 d, 7 d time points higher than the sham group(P <0 05.); brain clear microemulsion administration group AQP-4 expression were lower than at the corresponding time points ICH model group(P <0 05.);4. The impact on neuronal apoptosis after intracerebral hemorrhageIn this research, the change of expression about Pro-apoptosis factors Caspase-3 and anti-apoptopic protein Bcl-2 were determined by immunohistochemistry staining technique after Nao-Qing microemulsion administration of intracerebral hemorrhage rats model through a special interval, such as 1 day, 3 days, 5 days, 7 days. Result has shown that in normal brain tissue within the cerebral cortex, Caspase-3/Bcl-2 were weakly positive expressed while the number of Caspase-3 positive neuron increased significantly during the first day after intracerebral hemorrhage(p<0.05); reaching the peak in the third day with lots of positive cell nucleus and cytoplasm is tan, which was revealed a significant difference compared with sham-operation group(P<0.01);Conclusions1. Nao-Qing microemulsion can effective against brain injury in the central nervous system caused by cerebral hemorrhage.2. Nao-Qing microemulsion through AQP-4 / MMP-9 can effectively improve cerebral edema caused by brain hemorrhage.3. Nao-Qing microemulsion via Caspase-3 / Bcl-2 can effectively improve brain cell apoptosis caused by brain bleeding.This study has proved the mechanism of Nao-Qing microemulsion for treatment of intracerebral hemorrhage.This investigation established the foundation for developing the Nao-Qing Microemulsion,and provided references for the future clinical application.