Experimental Study Of AQP4, Kir4.1, Na+, K+-ATPase And NGB In Intracerebral Hemorrhage In Human

Intracerebral hemorrhage (ICH) is a common disease in the nervous system characterized by high mortality, relapse rate and unability. The incidence of cerebrovascular disease tended to increase year by year as people's living standards improved, and more younger. It is different from cerebral ischemia diseases, because we don't have obviously breakthrough in its therapy methed in recent years. It becomes a great burden to patients, families and society. In this paper, we observed the pathological changes of tissues around lesion.we studied the characteristics and the mechanism from cell, molecular, genetic level in order to find the appropriate treatment to reduce the degree of edema and reduce the morbidity and mortality and improve the prognosis of cerebral hemorrhage.Objective:One of the hot topic in recent years is about the molecular mechanisms of brain edema in neuroscience research.In this study, we use the surrounding tissue of intracerebral hemorrhage as specimen which we extract from surgery, we observe the change about AQP4, Kir4.1, Na+, K+-ATPase and NGB in order to provide a theoretical treatment and a new targets of cerebral hemorrhage for the future.Methods:we observed the pathological changes of brain tissue around lesion stained by HE method under light microscope.Expression of AQP4, Kir4.1, Na+, K+-ATPase, NGB and these were investigated by immunohistochemistry at each time point and PCR method was used to detect the expression of AQP4 and Kir4.1, then we use Western-blot method to detect the Kir4.1 protein expression. At last, we detected brain water content and Na+, K+-ATPase activity.Results:The pathological changes under light microscope:In comtrol group, The cells were closely aligned with each other, it contained nerve cells and glial cells; At 6h after haematoma, the neurocells are swelling and brain tissues around lesion became loose and matrix had the slight edema; At 6h-24h after haematoma, the change became obvious and a various sizes of vacuoles between cells.which became obvious at 3d with the broken and necrosis of nerve cells, there were a lot of malacia and edema lesion, there were still a lot of degeneration and necrosis of nerve cells and a lot of proliferation of glial cells.The pathological changes under electron microscope:In comtrol group, membrane neuronal cells was integrite, nucleolus was clear, and organelle morphology was clear in neuronal cells, tight junction (TJ) were exist between vascular endothelial cells. Within 6h after haematoma,the membrane and nucleolus became swelling but TJ still exist. At 6h-24h after haematoma, the swelling was obvious, and TJ was opening. At 24h-3d after haematoma, neurons and glial cells were dissolved, mitochondrial was swollen, deformated and shrink, cytoplasmic was vacuolized, nuclear chromatin was marginated, cell organelles structure was disorder and unclear, TJ opened variously.The results of immunohistochemistry:The expression of AQP4, Kir4.1 and NGB increased and the expression and activity of Na+, K+-ATPase decreased in peripheral edema after intracerebral hemorrhage. Immunoreactive cells were mainly distributed in the cytoplasm of glial cells and neurons. There were a small amount of positive cells in the control group.the staining of cells surrounding blood vessels is most evident, we can see a small amount of positive expression of endothelial cells.PCR results indicated that the products electrophoresis all were positive expressed at 190bp after the target gene of AQP4 and Kir4.1 by PCR amplification. AQP4mRNA and Kir4.1 mRNA all were highly expressed in hemorrhage group than in comtrol group. The number of AQP4 and Kir4.1 expression increased at 6h-24h, peaked at 24h-3d,then decreased gradually.Western-blot results indicated that the products electrophoresis all were positive expressed at 42.5kD after the target gene of AQP4 by Western-blot amplification. AQP4 protein were highly expressed in hemorrhage group than in comtrol group. The number of AQP4 expression increased at 6h-24h, peaked at 24h-3d, then decreased gradually.Conclusions:The expression of AQP4, Kir4.1 and NGB increased and the expression and activity of Na+, K+-ATPase decreased in peripheral edema after intracerebral hemorrhage.and the severity of hypertension was related to water content of brain, AQP4, and Na+, K+-ATPase, the treatment for the future provide a theoretical basis for cerebral hemorrhage and a new direction This experimental study provided a therapeutic foundation for clinical treatment ICH in future.