The Research Of Mechanism Of Activated Protein C In White Matter Damage After Intrauterine Inflammation Exposure

Objective White matter damage (WMD) is an important risk factor for nervous systemsequela among premature infants. A great many epidemiology and laboratory research haverecently shown that WMD is closely related to intrauterine infection or inflammation.However, effective precaution and treatment are still under investigation. Given the factthat the activated protein C (APC) has a strong inhibitory effect on inflammation, this studytested the efficiency of APC on preventing inflammatory response of in vitro culturedmicroglia. For provide theoretical and practical evidences for a safe and effectivetherapeutic precept of WMD in preterm infants.Methods The microglia cells were isolated from the brain of newborn Sprague Dawley (SD)mice, they were purified by the differential attaching method, and identified using IBA1staining. The microglia cells were randomly divided into negative control group (treatedwith PBS), inflammatory group (treated with lipopolysaccharide, LPS), and LPS+APCintervention group (treated with LPS, followed by APC)，APC intervention group(treatedwith APC). The morphology of all the groups were observed under microscope, and theexpression of TNF-α and PAR-1were determined by immunofluorescence staining. TheIntegrated optical density (IOD) was used to quantify the intensity of the fluorescence.Results Primary cultured microglia is successful. Given the LPS evoked by stimulation ofinflammation, microglia activation performance form. On the contrary, the addition of APCcan reverse the morphological change caused by LPS. The inflammatory factor TNF-α was greatly upregulated after LPS treatment (P<0.01), while APC intervention can decrease itsexpression to a level comparable to negative control group and APC intervention group(P>0.05). The protective acceptor PAR-1was not affected under exposure to inflammatorygroup, negative control group, and APC intervention group (P>0.05), while APCintervention can significantly increase its expression (P<0.05).Conclusions The microglia cells were successfully isolated and purified. The LPS caninduce an inflammatory response in microglia cells. APC intervention can reverse the effectof LPS and inhibit the inflammatory response. The protective effect might be partiallythrough the upregulation of PAR-1expression. This study provide preliminary evidencesthat APC might be a novel agent that can be administered to gravida with intrauterineinfection/inflammation to decrease the risk of WMD of premature infants...