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首页> 中文期刊> 《器官移植》 >扶芳藤含药血清对大鼠胰岛细胞的保护机制

扶芳藤含药血清对大鼠胰岛细胞的保护机制

         
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Objective To investigate the protective effect and mechanism of serum containing Euonymus fortunei on the rat pancreatic islet cells. Methods Forty male SD rats were randomly divided into 5 groups (n=8 in each group), including the control group (normal rat islet cells were cultured with normal rat serum), ischemic preconditioning group (abdominal aorta was blocked first and then re-opened before the pancreas was obtained, and the pancreatic islet cells were cultured with normal rat serum), Euonymus fortunei treatment group (normal rat islet cells were cultured with rat serum containing Euonymus fortunei), Euonymus fortunei group and blank group (normal rats were administered orally with Euonymus fortunei extract or distilled water for the preparation of rat serum). Diphenylthiocarbazone (DTZ) staining was utilized to observe and calculate the quantity of islets. Acridine orange (AO)/propidium iodide (PI) staining was adopted to calculate the survival rate of islet cells. The insulin release experiment was performed to calculate the stimulation index (SI) and evaluate islet cell function. The concentration of glutathione (GSH) and nitric oxide (NO) in islet cells was detected using GSH and NO kits. The expression level of inducible nitric oxide synthase (iNOS) messenger RNA (mRNA) was quantitatively measured by reverse transcription polymerase chain reaction (RT-PCR). Results Islet cells were observed in specifically scarlet color after DTZ staining. The quantity of islet cells did not significantly differ among different groups (all P>0.05). Along with the prolongation of culture time, the activity of islet cells in each group was gradually decreased. At 72 h after isolation and culture, compared with the control group, the survival rate of the cells was significantly higher in the Euonymus fortunei treatment group (P<0.05). The insulin release test results demonstrated that compared with the control group, the SI of the ischemic preconditioning and Euonymus fortunei treatment groups was significantly increased (both P<0.05). Compared with the control group, the GSH contents of pancreatic islet cells in the ischemic preconditioning and Euonymus fortunei treatment groups were considerably enhanced, the NO content was significantly decreased, and the expression level of iNOS mRNA was significantly down-regulated (all P<0.05). Conclusions Euonymus fortunei can increase the survival rate of islet cells and enhance the function of pancreatic islets by increasing the level of GSH, down-regulating the expression of iNOS and decreasing the NO production.%目的 探讨扶芳藤含药血清对大鼠胰岛细胞的保护作用及其机制.方法 将40只雄性SD大鼠随机分为5组,每组8只,包括对照组(正常大鼠胰岛细胞予正常大鼠血清培养)、缺血预处理组(获取胰腺前阻断腹主动脉再开放,胰岛细胞予正常大鼠血清培养)、扶芳藤治疗组(正常大鼠胰岛细胞予大鼠扶芳藤含药血清培养)、扶芳藤组和空白组(正常大鼠分别予扶芳藤提取物或蒸馏水灌胃,用于制备大鼠血清).采用双硫腙(DTZ)染色法观察并计算胰岛数量;采用吖啶橙(AO)/碘化丙啶(PI)染色法,计算胰岛细胞存活率;采用胰岛素释放实验计算刺激指数(SI)来评价胰岛细胞功能;采用谷胱甘肽(GSH)及一氧化氮(NO)试剂盒测定胰岛细胞内GSH、NO含量;采用逆转录聚合酶链反应(RT-PCR)法检测诱导型一氧化氮合酶(iNOS)信使核糖核酸(mRNA)的表达水平.结果 经DTZ染色后胰岛细胞呈特异性猩红色,各组间胰岛细胞数量比较,差异无统计学意义(均为P>0.05).随培养时间延长,各组胰岛细胞活性均逐渐降低,分离培养72 h后,与对照组比较,扶芳藤治疗组细胞存活率更高,差异有统计学意义(P<0.05).胰岛素释放实验检测结果提示,与对照组相比,缺血预处理组及扶芳藤治疗组的SI均升高,差异均有统计学意义(均为P<0.05).与对照组比较,缺血预处理组及扶芳藤治疗组胰岛细胞的GSH含量均升高、NO含量均降低、iNOS mRNA表达水平均降低,差异均有统计学意义(均为P<0.05).结论 扶芳藤能通过提高GSH、降低iNOS表达及NO产生,提高胰岛细胞存活率,增强胰岛功能.

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