【摘要】 目的 探讨腹水引起的腹内高压对肝硬化小鼠肺组织水通道蛋白1（AQP1）和水通道蛋白5（AQP5）表达的影响。 方法 雄性美国癌症研究所（Institudo of Cancer Reseach，ICR）小鼠50只，随机取10只作正常对照组（腹压0 cm H2O，1 cm H2O=0.098 kPa），其余40只用四氯化碳建立肝硬化小鼠模型，并随机分为4组:肝硬化（腹压0 cm H2O）组、肝硬化（腹压5 cm H2O）组、肝硬化（腹压10 cm H2O）组、肝硬化（腹压20 cm H2O）组，通过腹腔注射不同量的白蛋白生理盐水形成不同的腹压，并维持腹压24 h后取肺组织行病理、免疫组织化学、肺湿/干比值及实时荧光定量PCR检测AQP1和AQP5 mRNA表达量。 结果 与正常对照小鼠相比，肝硬化小鼠肺AQP5、AQP1表达明显下降（Plt;0.05）；肝硬化小鼠随着腹内压的升高，肺湿/干比值升高，AQP5、AQP1表达相应增加（Plt;0.05）。 结论 肝硬化可以影响肺AQP1、AQP5的表达；肝硬化小鼠随着腹内压的升高，AQP1、AQP5表达相应增加，并与肺水肿的严重程度密切相关。【Abstract】 Objective To investigate the role of intra-abdominal hypertension caused by ascites on the expression of Aquaporin (AQP) 1 and AQP 5 in the lung of cirrhotic mice. Methods We randomly chose 10 from 50 male Institude of Cancer Research (ICR) mice to form the control group [intra-abdominal pressure (IAP)=0 cm H2O, 1 cm H2O=0.098 kPa]. The model of cirrhosis were prepared by subcutaneous injection of carbon tetrachloride for the rest 40 mice which were then randomly divided into 4 groups: cirrhosis (IAP=0 cm H2O) group, cirrhosis (IAP=5 cm H2O) group, cirrhosis (IAP=10 cm H2O) group, and cirrhosis (IAP=20 cm H2O) group. Saline with different volume of albumin was injected into the peritoneum of each mouse in order to form different IAP. After 24 hours, analysis of pathology, immunochemistry and wet/dry ratio was done for the lungs of these mice; and the expression of AQP1 and AQP5 at the protein and mRNA levels were analyzed by IHC and qRT-PCR. Results Compared with the normal mice, the expression of AQP1 and AQP5 in lungs of cirrhotic mice were significantly lower (Plt;0.05). Both the lung wet/dry ratio and the expression of AQP1 and AQP5 raised with the increase of IAP. Conclusion Cirrhosis can affect the expression of AQP1 and AQP5 in lungs. The expression of AQP5 and AQP1 in lungs of cirrhotic mice increases with the increase of IAP, which is also closely correlated with the severity of pulmonary edema.
ObjectiveTo explore the effect and mechanism of directive differentiation of microglia by SN50 on hypoxia-caused neurons injury in mice.MethodsThe microglia were isolated and purified from brain tissue of new-born BALB/c mice through differential velocity adherent and vibration technique. The quantity of the microglia was identified by immunofluorescence staining of inducible nitric oxide synthetase (iNOS) and ionized calcium binding adapter molecule 1 (Iba1) and real-time fluorescence quantitative PCR (qRT-PCR) for special expression genes [iNOS, CD32, and interlenkin 10 (IL-10)]. Then the microglia were cultured with SN50, and the expressions of nuclear factor κB (NF-κB), differentiation-related genes (iNOS, CD11b, IL-10, and CD206), and apoptosis were detected by Western blot, qRT-PCR, and flow cytometry, respectively. The hypoxia model of neuron was established, and the cell apoptosis was evaluated by MTT after 0, 2, 6, 12, 24, and 48 hours of anoxic treatment. The apoptosis related markers (Bcl-2 and Caspase-3) were measured by Western blot and flow cytometry. In addition, the neurons after anoxic treatment were co-cultured with SN50 treated microglia (experimental group) and normal microglia (control group) for 24 hours. And the cell viability and apoptosis related markers (Bcl-2 and Caspase-3) were also measured.ResultsImmunofluorescence staining and qRT-PCR analysis showed that the cells expressed the specific proteins and genes of microglia. Compared with the normal microglia, the relative expressions of NF-κB protein and iNOS and CD11b mRNAs in the microglia treated with SN50 significantly decreased (P<0.05), the relative expressions of IL-10 and CD206 mRNAs significantly increased (P<0.05), and the cell apoptosis rate had no significant change (P>0.05). Compared with the normal neurons, the cell viability, the relative expressions of Bcl-2 and Caspase-3 proteins after anoxic treatment significantly decreased (P<0.05), while the relative expressions of cleaved-Caspase-3 protein and cell apoptosis rate of neurons significantly increased (P<0.05). In the co-culture system, the cell viability, the relative expressions of Bcl-2 and Caspase-3 proteins were significantly higher in experimental group than those in control group (P<0.05), while the relative expressions of cleaved-Caspase-3 protein and cell apoptosis rate were significantly lower in experimental group than those in control group (P<0.05).ConclusionSN50 can induce the microglia differentiation into M2 type through NF-κB pathway. The SN50-induced microglia can protect neurons from hypoxic injury.
【摘要】 目的 探讨海洋肽对恶性肿瘤化学治疗（简称化疗）患者营养状况和免疫功能的影响。 方法 依照纳入排除标准选取2010年3－11月66例恶性肿瘤化疗患者，随机分为试验组和对照组，每组各33例。在正常饮食基础上，试验组和对照组分别服用海洋肽制剂和乳清蛋白制剂21 d，进行肝肾功能、营养状况及免疫指标的测定。 结果 干预前后两组肝肾功及血脂指标差异无统计学意义（Pgt;0.05），且均在正常范围内。试验组干预后体质指数（body mass index，BMI）、上臂围、上臂肌围、总蛋白、白蛋白、球蛋白、前白蛋白（prealbumin，PA）、转铁蛋白较干预前升高有统计学意义（Plt;0.05），而血红蛋白和三头肌皮褶厚度干预前后比较差异无统计学意义（Pgt;0.05）；对照组干预前后各指标差异均无统计学意义（Pgt;0.05）；试验组BMI、PA的前后差值较对照组高（Plt;0.05），而其他指标差值在两组间差异无统计学意义（Pgt;0.05）。两组在干预前后组内比较及组间免疫指标差值比较，差异均无统计学意义（Pgt;0.05）。 结论 海洋肽作为部分氮源应用于恶性肿瘤化疗患者，对患者的内脏蛋白、人体测量等均有一定的营养改善作用，但对免疫功能的影响不明显，尚待进一步研究。【Abstract】 Objective To investigate the effects of marine peptide on the nutritional status and immune function in malignant tumor patients undergoing chemotherapy. Methods According to inclusive and exclusive criteria, 66 malignant tumor patients undergoing chemotherapy from March 2010 to November 2010 were randomized into study group and control group with 33 patients in each group. The patients in the study group were given marine collagen peptide whey protein while those in the control group were given whey protein for 21 days. Liver and kidney function, nutritional status and immune function were observed before and after intervention. Results Liver, kidney function and blood lipids of all the patients were within normal range, and were not significantly different between the two groups before and after intervention (P＞0.05). After intervention, body mass index (BMI), arm circumference, arm muscle circumference, total protein, albumin, globulin, prealbumin (PA), transferring protein of the study group were significantly increased (P＜0.05), but hemoglobin and triceps skinfold thickness had no significant changes (P＞0.05). There was no significant difference of the above parameters in the control group before and after intervention (P＞0.05). The magnitude of change of PA and BMI before and after intervention were higher in the study group than those in the control group (P＜0.05), while the magnitude of change of other parameters before and after intervention showed no significant difference between the two groups (P＞0.05). The immune function showed no significant change in both groups before and after intervention (P＞0.05), and it was also not significantly different between the two groups. Conclusion As part of dietary nitrogen sources, marine peptide can significantly improve nutritional status, including visceral protein and anthropometry in malignant tumor patients undergoing chemotherapy, but it has no significant effect on immune function, which should be further studied in detail.