嘌呤受体P2X7抑制剂BBG对异氟烷麻醉致老年大鼠认知功能障碍的影响

Effects of P2X7 receptor antagonist BBG on cognitive dysfunction of elderly rats induced by isoflurane anesthesia

目的观察嘌呤受体P2X7阻断剂BBG对异氟烷麻醉致大鼠认知功能障碍的影响,初步探讨P2X7受体在其中的作用机制。方法雄性18月龄SD大鼠100只随机分为C组(对照组)、BBG组、ISO组(异氟烷组)和BBG+ISO组,每组25只。C组和ISO组腹腔注射0.9%生理盐水,BBG组和BBG+ISO组腹腔注射BBG溶液50 mg·kg^(-1)·d^(-1),连续3 d。第4 d,C组和BBG组持续吸入100%O26 h;ISO组和BBG+ISO组持续吸入1.4%异氟烷麻醉6 h,以100%O2作为载气。24 h后行Moris水迷宫实验观察大鼠认知功能变化,免疫荧光染色法观察Iba-1染色评估大鼠海马区小胶质细胞激活程度,酶联免疫吸附(ELISA)法检测大鼠海马组织中白细胞介素-1β(IL-1β)和IL-6浓度。Sigmaplot 12.0统计学软件进行分析,组间比较采用单因素方差分析,组内比较采用重复测量设计的方差分析,P<0.05为差异有统计学意义。结果与C组比较,ISO组行为学训练第3~5天逃避潜伏期明显延长,探索时间明显缩短(P<0.05),海马CA3区Iba-1阳性染色面积明显更高(P<0.01),海马IL-1β和IL-6含量明显高于其它3组(P<0.01)。与ISO组比较,BBG+ISO组训练第3~5天逃避潜伏期缩短,探索时间延长(P<0.05)。海马CA3区Iba-1阳性面积染色明显更低(P<0.05),海马IL-1β和IL-6含量更低(P<0.01)。结论P2X7受体阻断剂BBG可改善异氟烷麻醉引起的老年大鼠认知功能障碍,其作用机制可能是BBG抑制海马区域小胶质细胞激活,进而降低海马区域炎性反应,改善认知功能。

Objective To observe the effects of P2X7 receptor antagonist BBG on cognitive dysfunction of elderly rats in- duced by isoflurane anesthesia, and discuss the possible mechanism in these effects. Methods 100 Male SD rats （18 months old） were randomly divided into control group, BBG group, isoflurane （ISO） group, and ISO＋BBG group, 25 rats per group. BBG group and ISO＋BBG group were injected 50 mg·kg^-1·d^-1 introperitonealy for 3 days, control group and ISO group were injected saline with same volume. 1.4% Isoflurane anesthesia were performed for 6h on day 4. Cognitive function were assessed by Moils water maze 24 h later. Microglia Iba-1 staining were measured by immunofluorescent staining, enzyme-linked immuno sorbent assay （ELISA） were used to quantify the concentration of interleukin-1β （ IL-1β） and IL-6 in hippocampus. Sigmaplot 12.0 statistical software was used to analyze the data by One-way ANOVA or RM ANOVA. P〈0.05 was considered statistical differences. Results Compared with control group, the escape latency was prolonged, the time of staying at the original platform quadrant was shortened from day 3 to day 5, the positive area of Iba-1 staining at hippocampus CA3 area, the concentration of IL-1β and IL-6 were all significantly increased in ISO group （P〈0.05）. Compared with ISO group, the escape latency was shortened, the time of staying at the original platform quadrant was prolonged, the positive area of Iba-1 staining at hippocampus CA3 area, the concentration of IL-1β and IL-6 were sig- nificantly decreased in ISO＋BBG group （P〈0.05）. Conclusion BBG can attenuate isoflurane anesthesia induced cognitive decline via inhibiting the activity of hippocampus microglia and neuroinflammation.