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Tian L.-Y.,University of Sichuan | Li S.-F.,Key Laboratory of Transplant Engineering | Zhou D.,University of Sichuan
Journal of Sichuan University (Medical Science Edition) | Year: 2010

Objective To develop a separation method for brain microendothelial cells with the comparison of other ones. Methods Twice enzymatic digestion and twice gradient centrifugation were applied to separate rat brain microendothelial cells. Then, immunomagnetic beads and Thyl. 1 antibody were used respectively to purify the cultured cells. Results Twice enzymatic digestion and twice gradient centrifugation could separate the cell successfully. High purification but low cell yield was obtained with immunomagnetic beads. The cells handled with Thyl. 1 antibody had both higher purify coefficient and higher yield. Conclusion The developed method could separate the brain microendothelial cells successfully.

Guo J.Y.,Key Laboratory of Transplant Engineering | Yang T.,Key Laboratory of Transplant Engineering | Sun X.G.,Key Laboratory of Transplant Engineering | Zhou N.Y.,Key Laboratory of Transplant Engineering | And 5 more authors.
Journal of Biomedical Science | Year: 2011

Background: Ischemic postconditioning (IPO) has been demonstrated to attenuate ischemia/reperfusion (I/R) injury in the heart and brain, its roles to liver remain to be defined. The study was undertaken to determine if IPO would attenuate liver warm I/R injury and its protective mechanism. Methods. Mice were divided into sham, I/R, IPO+I/R (occlusing the porta hepatis for 60 min, then treated for three cycles of 10 sec brief reperfusion consecutively, followed by a persistent reperfusion); L-NAME+ sham (L-NAME, 16 mg/kg, i.v., 5 min before repefusion); L-NAME+I/R; and L-NAME+ IPO. Blood flow of caudate and left lobe of the liver was blocked. Functional and morphologic changes of livers were evaluated. Contents of nitric oxide, eNOS and iNOS in serum were assayed. Concentration of eNOS, iNOS, malondialdehyde (MDA) and activity of superoxide dismutase (SOD) in hepatic tissue were also measured. Expressions of Akt, p-Akt and HIF-1 protein were determined by western blot. Expressions of TNF- and ICAM-1 were measured by immunohistochemistry and RT-PCR. Results: IPO attenuated the dramatically functional and morphological injuries. The levels of ALT was significantly reduced in IPO+I/R group (p < 0.05). Contents of nitric oxide and eNOS in serum were increased in the IPO+I/R group (p < 0.05). IPO also up-regulated the concentration of eNOS, activity of SOD in hepatic tissue (p < 0.05), while reduced the concentration of MDA (p < 0.05). Moreover, protein expressions of HIF-1 and p-Akt were markedly enhanced in IPO+I/R group. Protein and mRNA expression of TNF- and ICAM-1 were markedly suppressed by IPO (p < 0.05). These protective effects of IPO could be abolished by L-NAME. Conclusions: We found that IPO increased the content of NO and attenuated the overproduction of ROS and I/R-induced inflammation. Increased NO contents may contribute to increasing HIF-1 level, and HIF-1 and NO would simultaneously protect liver from I/R injury. These findings suggested IPO may have the therapeutic potential through Akt-eNOS-NO-HIF pathway for the better management of liver I/R injury. © 2011 Guo et al; licensee BioMed Central Ltd.

Xiao Z.,Key Laboratory of Transplant Engineering | Shan J.,Key Laboratory of Transplant Engineering | Li C.,Key Laboratory of Transplant Engineering | Luo L.,Key Laboratory of Transplant Engineering | And 5 more authors.
American Journal of Nephrology | Year: 2013

Background/Aims: Chronic cyclosporine A (CsA) nephrotoxicity (CCN) is an important cause of chronic renal dysfunction with no effective clinical intervention. To further elucidate the mechanisms of renal cell apoptosis in CCN, all relevant in vivo studies on this subject were analyzed. Methods: We searched for in vivo studies on the mechanisms of CsA-induced renal cell apoptosis in Medline (1966-July 2010), Embase (1980-July 2010) and ISI (1986-July 2010). The studies were evaluated for their quality according to a set of in vivo standards, data extracted according to PICOS, and then synthesized. Results: Renal cell apoptosis was an important feature of CCN and an important factor of renal dysfunction. First, CsA could upregulate Fas/Fas ligand, downregulate Bcl-2/Bcl-XL, and increase caspase-1 and caspase-3. Second, it could induce oxidative stress and damage the antioxidant defense system. Third, it could increase endoplasmic reticulum stress protein in a dose- and time-dependent manner. Fourth, CsA could impair the urine concentration and decrease the expression of hypertonicity-induced genes. Fifth, CsA-induced renal cell apoptosis was significantly decreased by blocking the angiotensin II type 1 receptor using losartan. Conclusions: The in vivo mechanisms for CCN are more complex than those found in vitro. CsA can induce renal cell apoptosis using five pathways in vivo and activated caspases might be the ultimate intersection of these pathways and the common intracellular pathway mediating apoptosis. These data provide new potential points for intervention and need to be confirmed by further studies. Copyright © 2012 S. Karger AG, Basel.

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