探讨丁苯酞（BNP）对七氟醚诱导神经细胞损伤的影响及其可能的机制。

取胎鼠海马组织，分离培养神经细胞，分为对照组（正常神经细胞）、七氟醚组（3%七氟醚通气）、低剂量BNP组（1 μmol/L的BNP处理6 h+3%七氟醚通气）、中剂量BNP组（5 μmol/?L的BNP处理6 h + 3%七氟醚通气）以及高剂量BNP组（10 μmol/L的BNP处理6?h + 3%七氟醚通气），采用MTT法检测细胞生存率，采用流式细胞术检测细胞凋亡情况，采用Western bolt法检测各组细胞Bcl-2、Bax以及Caspase-3蛋白表达情况，生存率、凋亡率比较采用t检验或方差分析。

24、48和72 h，七氟醚同期处理各组细胞生存率较对照组明显降低[对照组（88.31±8.20）%，（80.42±9.04）%，（75.63±7.84）%；七氟醚组（71.53±6.95）%，（61.35±7.38）%，（45.40±5.42）%，t = 3.491、3.654、7.092，P均< 0.001；低剂量BNP组（72.41±8.42）%，（67.22±6.46）%，（50.11±5.71）%，t = 3.025、2.657、17.411，P?= 0.016、0.027、0.000；中剂量BNP组（76.68±5.95）%，（72.53±5.10）%，（54.58±6.73）%，t = 2.567、2.030、4.556，P = 0.028、0.034、0.000；高剂量BNP组（79.32±6.44）%，（75.68±5.47）%，（60.09±4.28）%；t = 2.183、2.048、3.890，P = 0.023、0.036、0.000]，而BNP各处理组细胞生存率明显高于单纯七氟醚处理组[七氟醚组（71.53±6.95）%，（61.35±7.38）%，（45.40±5.42）%，低剂量BNP组（72.41±8.42）%，（67.22±6.46）%，（50.11±5.71）%，t = 1.180、2.038、2.317，P?= 0.216、0.030、0.019；中剂量BNP组（76.68±5.95）%，（72.53±5.10）%，（54.58±6.73）%，t?= 2.180、2.787、2.376，P = 0.028、0.017、0.020；高剂量BNP组（79.32±6.44）%，（75.68±5.47）%，（60.09±4.28）%，t = 2.206、3.488、4.756，P = 0.022、0.000、0.000]，且随着BNP浓度的升高，细胞生存率明显升高[低剂量BNP组（72.41±8.42）%，（67.22±6.46）%，（50.11±5.71）%；中剂量BNP组（76.68±5.95）%，（72.53±5.10）%，（54.58±6.73）%；高剂量BNP组（79.32±6.44）%，（75.68±5.47）%，（60.09±4.28）%；F = 2.182、2.491、2.384，P = 0.020、0.024、0.022]。采用流式细胞技术检测各组神经细胞凋亡情况，采用七氟醚通气处理的各组神经细胞凋亡率均显著高于对照组[七氟醚组（67.49±7.92）%，低剂量BNP组（60.72±8.33）%，中剂量BNP组（44.95±7.21）%，高剂量BNP组（31.86±6.50）%，对照组（19.42±4.58）%，F = 6.583，P < 0.001]，而采用BNP处理的各组神经系统凋亡率明显低于七氟醚组（P < 0.05），且随着BNP剂量的增加，凋亡率明显降低（P < 0.05）。七氟醚通气各组细胞Bcl-2表达较对照组降低，而BNP处理各组细胞Bcl-2表达较七氟醚组有所升高，且随着BNP剂量增加Bcl-2表达增加；七氟醚通气各组细胞Bax以及Caspase-3蛋白表达较对照组增加，而BNP处理各组细胞Bax以及Caspase-3蛋白表达较七氟醚组有所降低，且随着BNP浓度的升高Bax以及Caspase-3蛋白表达降低。

七氟醚可诱导神经细胞凋亡，从而对神经系统造成损伤，而BNP则可有效抑制七氟醚诱导的神经细胞凋亡，对神经系统损伤具有保护作用，其可能的机制与促进Bcl-2蛋白表达，抑制Bax、Caspase-3蛋白表达有关。

To investigate the effect of butylphthalide on nerve cell injury induced by sevoflurane and its possible mechanism.

Nerve cells from fetal rat hippocampus were cultured and divided into the control group (normal nerve cells), sevoflurane group (3% sevoflurane ventilation), low dose BNP group (BNP 1 μmol/L 6 h+3% sevoflurane ventilation), intermidate dose BNP group (BNP 5 μmol/L 6 h+3% enlfurane ventilation ) and high dose BNP group (BNP 10 μmol/L 6 h+3% sevoflurane ventilation). The survival of cells was detected by MTT method, the cell apoptosis was detected by flow cytometry and Bcl-2, Bax and Caspase-3 protein expression were evaluated by Western bolt.

At 24 h, 48 h and 72 h, sevoflurane cell survival rates were significantly lower than that of the control group (Control group:(88.31±8.20)%, (80.42±9.04)%, (75.63±7.84)%; Sevo group:(71.53±6.95)%, (61.35±7.38)%, (45.40±5.42)%. t = 3.491, 3.654, 7.092, P < 0.001. In the low dose BNP group, survival rates were: (72.41±8.42)%, (67.22±6.46)%, (50.11±5.71)%. t = 2.567, 2.030, 4.556, P = 0.028, 0.034, 0.000.In the moderate dose BNP group, survival rates were: (76.68±5.95)%, (72.53±5.10)%, (54.58±6.73)%. t = 2.567, 2.030, 4.556, P?= 0.028, 0.034, 0.000. High dose BNP group: (79.32±6.44)%, (75.68±5.47)%, (60.09±4.28)%, t = 2.183, 2.048, 3.890, P = 0.023, 0.036, 0.000). Cell survival rate of each treatment group was significantly higher than that of the sevoflurane group (Sevo group: (71.53±6.95)%, (61.35±7.38)%, (45.40±5.42)%. Low dose BNP group: (72.41±8.42)%, (67.22±6.46)%, (50.11±5.71) %. t = 1.180, 2.038, 2.317, P = 0.216, 0.030, 0.019. Moderate dose BNP group: (76.68±5.95)%, (72.53±5.10) %, (54.58±6.73) %. t = 2.180, 2.787, 2.376, P = 0.028, 0.017, 0.020. High dose BNP group: (79.32±6.44)%, (75.68±5.47)%, (60.09±4.28)%, t = 2.206, 3.488, 4.756, P = 0.022, 0.000, 0.000), and with the increasing of BNP concentration, cell survival rate was significantly increased (Low dose BNP group: (72.41±8.42)%, (67.22±6.46)%, (50.11±5.71)%. Moderate dose BNP group: (76.68±5.95)%, (72.53±5.10)%, (54.58±6.73)%. High dose BNP group: (79.32±6.44)%,(75.68±5.47)%, (60.09±4.28)%, F = 2.182, 2.491, 2.384, P = 0.020, 0.024, 0.022). Flow cytometry showed apoptosis of nerve cells treated with sevoflurane was significantly higher than the control group (Sevo group and the apoptosis was prevented by addition of BNP in a dose-dependent manner:(67.49±7.92)%, Low dose BNP group:(60.72±8.33)%, moderate dose BNP group: (44.95±7.21)%, High dose BNP group:(31.86±6.50)%, Control group:(19.42±4.58)%, F = 6.583, P < 0.001). The expression of Bcl-2 was lower in the sevoflurane group than the control group; BNP increased the expression of Bcl-2; sevoflurane treatment increased Bax and Caspase-3 protein expression, which was prevent by BNP.

Sevoflurane can induce the apoptosis of nerve cells, thereby causing damage to the nervous system, and butylphthalide can effectively inhibit the apoptosis of nerve cells induced by sevoflurane by promoting the expression of Bcl-2 protein and inhibiting Bax and Caspase-3 protein expression.