【摘要】 目的 探討腹水引起的腹內高壓對肝硬化小鼠肺組織水通道蛋白1（AQP1）和水通道蛋白5（AQP5）表達的影響。 方法 雄性美國癌癥研究所（Institudo of Cancer Reseach，ICR）小鼠50只，隨機取10只作正常對照組（腹壓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，并隨機分為4組:肝硬化（腹壓0 cm H2O）組、肝硬化（腹壓5 cm H2O）組、肝硬化（腹壓10 cm H2O）組、肝硬化（腹壓20 cm H2O）組，通過腹腔注射不同量的白蛋白生理鹽水形成不同的腹壓，并維持腹壓24 h后取肺組織行病理、免疫組織化學、肺濕/干比值及實時熒光定量PCR檢測AQP1和AQP5 mRNA表達量。 結果 與正常對照小鼠相比，肝硬化小鼠肺AQP5、AQP1表達明顯下降（Plt;0.05）；肝硬化小鼠隨著腹內壓的升高，肺濕/干比值升高，AQP5、AQP1表達相應增加（Plt;0.05）。 結論 肝硬化可以影響肺AQP1、AQP5的表達；肝硬化小鼠隨著腹內壓的升高，AQP1、AQP5表達相應增加，并與肺水腫的嚴重程度密切相關。【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 探討海洋肽對惡性腫瘤化學治療（簡稱化療）患者營養狀況和免疫功能的影響。 方法 依照納入排除標準選取2010年3－11月66例惡性腫瘤化療患者，隨機分為試驗組和對照組，每組各33例。在正常飲食基礎上，試驗組和對照組分別服用海洋肽制劑和乳清蛋白制劑21 d，進行肝腎功能、營養狀況及免疫指標的測定。 結果 干預前后兩組肝腎功及血脂指標差異無統計學意義（Pgt;0.05），且均在正常范圍內。試驗組干預后體質指數（body mass index，BMI）、上臂圍、上臂肌圍、總蛋白、白蛋白、球蛋白、前白蛋白（prealbumin，PA）、轉鐵蛋白較干預前升高有統計學意義（Plt;0.05），而血紅蛋白和三頭肌皮褶厚度干預前后比較差異無統計學意義（Pgt;0.05）；對照組干預前后各指標差異均無統計學意義（Pgt;0.05）；試驗組BMI、PA的前后差值較對照組高（Plt;0.05），而其他指標差值在兩組間差異無統計學意義（Pgt;0.05）。兩組在干預前后組內比較及組間免疫指標差值比較，差異均無統計學意義（Pgt;0.05）。 結論 海洋肽作為部分氮源應用于惡性腫瘤化療患者，對患者的內臟蛋白、人體測量等均有一定的營養改善作用，但對免疫功能的影響不明顯，尚待進一步研究。【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.