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Objective:To evaluate the effect of hydrogen on the immunosuppressive status of septic rats.Methods:SPF healthy adult male Sprague-Dawley rats, aged 7-8 weeks, weighing 220-260 g, were studied.This study was performed in two parts.Part Ⅰ The rats were divided into 2 groups: sepsis group (Sep group, n=36) and sham operation group (Sham group, n=12). The model of sepsis was established by cecal ligation puncture in anesthetized rats.The histocompatibility DR antigen (HLA-DR)/CD14 + monocyte level in peripheral blood was detected by flow cytometry immediately after CLP and at 1, 2, 3 and 4 days after CLP.The establishment of sepsis-induced immunosuppression model was considered successful when the levels of HLA-DR/CD14 + monocyte in peripheral blood were <30%.Part Ⅱ Twelve rats with sepsis-induced immunosuppression were randomly selected and divided into 2 groups ( n=6 each) by a random number table method: sepsis immunosuppression group (Sep-IS group), sepsis immunosuppression plus hydrogen treatment group (Sep-IS+ H group). Another 12 rats were selected and divided into 2 groups ( n=6 each) by a random number table method: sham operation group (Sham group) and sham operation plus hydrogen group (Sham+ H group). In Sep-IS+ H group, 67% hydrogen was inhaled for 1 h starting from the time point immediately after successful establishment of sepsis-induced immunosuppression and from 6 h after establishment, and 67% hydrogen was inhaled for 1 h at the corresponding time points in Sham+ H group.The levels of helper T lymphocytes 17 (Th17 cells), regulatory T lymphocytes (Treg cells) and HLA-DR/CD14 + monocyte in peripheral blood were determined by flow cytometry immediately after the end of hydrogen inhalation mentioned above (T 0, T 1) and at 12 h after establishing the model (T 2). Results:Part Ⅰ Compared with Sham group, the levels of HLA-DR/CD14 + monocyte in peripheral blood were significantly decreased at 1-4 days after CLP in Sep group ( P<0.05). Part Ⅱ Compared with Sham group, the level of HLA-DR/CD14 + monocytes in peripheral blood was significantly decreased, and the levels of Treg and Th17 cells were increased at each time point in Sep-IS and Sep-IS+ H groups ( P<0.05), and no significant change was found in the parameters mentioned above in Sham+ H group ( P>0.05). Compared with Sep-IS group, the level of HLA-DR/CD14 + monocytes in peripheral blood was significantly increased at T 1, 2, the levels of Th17 cells were increased at T 2, and the levels of Treg cells were decreased at T 1, 2 in Sep-IS+ H group ( P<0.05). Conclusion:Hydrogen can improve the immunosuppressive state of septic rats.
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Objective To evaluate the efficacy of berberine in preventing brain injury induced by hepatic ischemia-reperfusion ( I∕R) and the relationship with glycogen synthase kinase 3 beta ( GSK-3β) activity in mice. Methods Forty-eight healthy clean-grade male C57BL∕6 mice, weighing 20-25 g, were randomized into 3 groups (n=16 each) using a random number table method: sham operation group (S group), hepatic I∕R group (I∕R group) and berberine group (B group). The model of 70％ liver I∕R injury was established by clamping the portal vein and hepatic artery supplying left and middle lobes of the liver in mice anesthetized with 2％ pentobarbital sodium 40 mg∕kg. In B group, berberine 50 mg∕kg was adminis-tered through a gastric tube once a day for 7 consecutive days starting from 7 days before operation. The e-qual volume of normal saline was given instead in S group and I∕R group. The mice were sacrificed at 6 h af-ter reperfusion, brains were removed and hippocampal tissues were harvested for microscopic examination of pathological changes ( with a light microscope) and for determination of cell apoptosis ( by TUNEL) , super-oxide dismutase ( SOD ) activity ( by xanthine oxidase method ) , malondialdehyde ( MDA ) content ( by thiobarbituric acid colorimetric method) , expression of phosphorylated GSK-3β ( p-GSK-3β) and GSK-3β(by Western blot), and opening of mitochondrial permeability transition pore (mPTP). The apoptosis in-dex and p-GSK-3β∕GSK-3β ratio were calculated. Results Compared with S group, the apoptosis index and MDA content were significantly increased, SOD activity and p-GSK-3β∕GSK-3β ratio were decreased, and mPTP opening was increased in I∕R and B groups ( P<0. 05) . Compared with group I∕R, the apoptosis index and MDA content were significantly decreased, SOD activity and p-GSK-3β∕GSK-3β ratio were in-creased, mPTP opening was decreased (P<0. 05), and the pathological changes of hippocampal tissues were significantly attenuated in B group. Conclusion Berberine can prevent the brain injury induced by he-patic I∕R, and the mechanism may be related to inhibiting GSK-3βactivity and thus inhibiting mPTP open-ing in mice.
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Objective To evaluate the effect of propofol on subunit 2B-containing N-methyl-D-aspartate receptor (NR2B)/calcium/calmodulin-dependent protein kinase Ⅱ alpha (CaMKⅡα) signaling pathway in brain injury induced by hepatic ischemia-reperfusion (I/R) in preadolescent mice.Methods Sixty-four healthy clean-grade C57BL/6 mice,aged 2 weeks,weighing 4-6 g,were randomized into 4 groups (n=16 each) using a random number table method:sham operation group (S group),hepatic I/R group (HI/R group),propofol control group (P group),and propofol plus hepatic I/R group (P+ HI/R group).The model of 70％ liver I/R injury was established by clamping the left and middle lobe vascular trunk in anesthetized mice.Propofol 20 mg/kg was intraperitoneally injected before operation at 30 min before establishing the model in P and P+HI/R groups.The equal volume of normal saline was given instead in P and P + HI/R groups.Eight mice of each group were sacrificed at 6h of reperfusion,hippocampal tissues were obtained for examination of pathological changes of hippocampal tissues and for determination of neuronal apoptosis (by TUNEL) and expression of NR2B,phosphorylated NR2B (p-NR2B),CaMKⅡα and phosphorylated CaMKⅡα (p-CaMKⅡα) (by Western blot).The remaining 8 mice in each group were used for Morris water maze test at 1 month after establishing the model.Apoptosis index was calculated.Results Compared with group S,the escape latency was significantly prolonged,the percentage of the time of staying at the original platform quadrant was decreased,the expression of p-NR2B and p-CaMKⅡα was up-regulated,and apoptosis index was increased in HI/R and P+HI/R groups (P＜0.05),and no significant change was found in the parameters mentioned above in group P (P＞0.05).Compared with group HI/R,the escape latency was significantly shortened,the percentage of the time of staying at the original platform quadrant was increased,the expression of p-NR2B and p-CaMKⅡα was down-regulated,and apoptosis index was decreased (P＜0.05),and the pathological changes of hippocampal tissues were significantly attenuated in group P+HI/R.Conclusion The mechanism by which propofol reduces brain injury induced by hepatic I/R may be related to inhibiting NR2B/CaMKⅡα signaling pathway in preadolescent mice.