Tanshinone, Blood Flux Of Tnf Alpha Induction Effect And Its Mechanism Of Aqp4 Expression

Cerebral edema is common pathological phenomenon of many neurological diseases, and a key factor of the deadly nerve injury cause these diseases later, for the treatment of cerebral edema, however, the application of traditional dehydrated drugs and can not significantly improve the prognosis and mortality. Therefore, in-depth study of the molecular mechanisms of cerebral edema water balance disorders found that the intervention more effective drugs and therapeutic targets for reducing brain disease mortality and improve the quality of life of patients has a very important significance. A large number of studies have found that AQP4-mediated transmembrane transport of water, is closely related with the formation of brain edema and lessons. Inflammatory factors TNF-a-mediated inflammatory response after brain injury to participate in the pathophysiological process of cerebral edema. In recent years, studies have shown that inflammatory factors through the regulation of AQPs expression is involved in tissue edema, our previous experiments also confirmed that TNF-a and cerebral edema and AQP4expression was positively correlated with an idea for the treatment of cerebral edema. Our previous study found promoting blood circulation by reducing the degree of inflammatory response to cerebral edema, according to Chinese medicine "blood related with water" theory, combined with the characteristics of the pathogenesis of cerebral edema, our current focus on traditional Chinese medicine on the TNF-a-induced expression of AQP4and its mechanism.Purpose1.Explore TNF-a whether can be induced AQP4expression in vitro cultured rat astrocytes.2.Explore the signaling pathway of TNF-a induced expression of AQP4.3.Explore traditional Chinese medicine on the TNF-a-induced expression of AQP4and its mechanisms, provide ideas and experimental evidence for the clinical treatment of cerebral edema.Method1.Cultured astrocytes, to determine the concentration of TNF-α, chose the concentration of TNF-a0,0.02,0.2,2,6,10ng/ml cultured cells6h for the Real-time PCR detection,and15h for Western blot analysis;2.Selection the peak concentration (2ng/ml) of TNF-a and cultured cells0,3,6,15,24h for Real-time PCR detection and0,6,15,24h for Western blot;3.PDTC pretrcatment30min and cultured cells with2ng/ml of TNF-a6h for the Real-time PCR,15h for Western blot analysis;4.The control group, TNF-a group, the group of tanshinone with low concentration (low concentration of tanshinone+TNF-α), the group of tanshinone with high concentration (high concentration of tanshinone-t TNF-α), the group of Xucsaitong with low concentration (low concentration of Xuesaitong+TNF-α), the group of Xuesaitong with high concentration (high concentration of Xuesaitong+TNF-α) cultured cells6h for the Real-time PCR and15h for Western blot analysis.Results1.TNF-a induces the expression of AQP4in a concentration-dependent manner. TNF-α induced expression of AQP4mRNA most obvious in2ng/ml and6ng/ml, the group of10ng/ml mRNA expression somewhat lower;TNF-a induced the expression of AQP4protein, The AQP4of TNF-a concentration at0.02ng/ml began to increase,2ng/ml reached peak,6ng/ml began to fall,and the10ng/ml group still higher than the control group.2.TNF-α induces the expression of AQP4in a time-dependent manner. TNF-a-induced expression of AQP4mRNA began at3h,6h reached a peak,15h began to fall and24h tends to normal. TNF-a-induced AQP4protein expression begins at6h,15h reached a peak,24h group began to drop, but still higher than normal levels.3.PDTC can block TNF-a-induced expression of AQP4. Compared with the control group, AQP4mRNA content of TNF-a group increased significantly at6h, with PDTC pretreatment30min and then added to TNF-α cultured for6h, AQP4mRNA no significant difference compared with the control group, compared with TNF-α significantly reduced. At15h, TNF-α group compared with the control group, the AQP4protein content was significantly increased, with PDTC pretreatment30min and then TNF-a cultured for15h, no significant difference compared the AQP4protein with the control group, than with TNF-a significantly reduced.4. Traditional Chinese medicine can inhibit the induction of TNF-a on AQP4. Compared with the TNF-a group, tanshinone group AQP4mRNA and protein were significantly reduced,AQP4expression of tanshinone high concentration group is no difference with the control group; compared with the TNF-a group, Xuesaitong group AQP4mRNA and protein were significantly reduced, Xuesaitong low concentration group AQP4expression compared with the control group is no difference.Conclusion1.TNF-a expression leads to cerebral edema by inducing AQP4, and this process of concentration and time-dependent manner.2.TNF-α induces the expression of AQP4through the NF-kappaB pathway.3.Tanshinone and xuesaitong induces cerebral edema by inhibiting inflammatory factors induceing AQP4.