DP2干预对铝负荷原代培养大鼠海马神经元的作用观察

摘要

Aim To establish primary cultured rat hip-pocampal neuron damage model induced by aluminum maltolate and study the effect of intervention for DP2 on primary cultured rat hippocampal neuron treated with aluminum overload. Methods The hippocampus was dissected out from fetal rat ( embryonic 18 d ) . After being cultured for 7 d, the hippocampal neuron was treated with Al( malt) 3 to establish the model of prima-ry cultured rat hippocampal neuron damage and mean-while treated with DP2 agonist DK-PGD2 and DP2 an-tagonist CAY10471, respectively. After treatment for 24 h, the cell viability was measured by MTT and LDH, Ca2+ fluorescence intensity. Neuronal pathomor-phology was observed by HE staining. Results The purity of hippocampal neuron was more than 95%. Compared with the control group, the number of hipp-ocampal neurons was reduced and neurons became chromatic agglutination and karyopyknosis in aluminum overload group. Treatment of aluminum caused a sig-nificant decrease in MTT value ( P<0. 01 ) and an in-crease in the LDH leakage rate (P<0. 01). The Ca2+fluorescence intensity significantly increased ( P <0. 01 ) in aluminum overload group. Compared with that of the aluminum overload group, treatment of DK-PGD2 , a selective DP2 agonist, significantly aggravated the primary cultured rat hippocampal neuron injury caused by aluminum overload accompanied with the significant decrease of MTT value ( P <0. 01 , P <0. 05 ) and an increase of the LDH leakage rate ( P<0. 01), significant increase of Ca2+ fluorescence inten-sity of neuron. Treatment of CAY10471, a selective DP2 antagonist, had opposite effects of DK-PGD2 . Conclusion The activation of DP2 can increase hipp-ocampal neural susceptibility to aluminum overload.%目的：建立麦芽酚铝致原代培养大鼠海马神经元损伤模型,探讨DP2干预对铝负荷原代海马神经元的作用。方法选取孕期18 d 左右的SD 大鼠,离体培养胎鼠海马神经元,d 7进行NSE 免疫组化鉴定,并给予Al(malt)3建立铝负荷致原代培养大鼠海马神经元损伤模型,同时分别给予DP2激动剂DK-PGD2和DP2拮抗剂CAY10471进行干预。继续培养24 h 后,检测各组海马神经元MTT 值、LDH 漏出率和 Ca2+荧光强度,HE 染色观察神经元病理形态变化。结果海马神经元纯度超过95%。与空白对照组比较,铝负荷模型组MTT 值明显降低(P <0．01)；LDH 漏出率明显升高(P <0．01)；Ca2+荧光强度明显增强(P <0．01)；神经元细胞数目明显减少,突起萎缩甚至消失,部分细胞核固缩。与铝负荷模型组比较,DP2激动剂DK-PGD2干预组MTT 值明显降低(P <0．01、P <0．05)；LDH 漏出率明显升高(P <0．01)； Ca2+荧光强度有增强趋势,但差异无显著性；海马神经细胞几乎全部核固缩、裂解。DP2拮抗剂CAY10471干预组MTT 值明显升高(P <0．01)；LDH 漏出率明显降低(P <0．01)； Ca2+荧光强度明显减弱(P <0．01)。海马神经细胞胞体、胞核明显,裂解细胞明显减少。结论 DP2激活表达可增加神经元对铝盐损伤的易感性。