摘要目的 观察川芎嗪(TMP)对氧诱导视网膜病变(OIR)中视网膜神经细胞凋亡的抑制作用.方法 48只C57BL/6新生幼鼠随机分为正常对照组、OIR对照组及OIR药物组,分别为18、18、12只.正常对照组在正常空气环境中饲养.OIR对照组、OIR药物组幼鼠于出生后7 d置于含氧体积分数(75±3)%的氧气箱中连续生活5 d;出生后12 d,幼鼠回到正常空气环境中饲养,建立OIR模型.出生后12～16d,OIR药物组按200 mg/kg的剂量腹腔注射TMP,1次/d;正常对照组、OIR对照组同时注射相同体积的含0.1%二甲基亚砜(DMSO)的生理盐水.出生后12、14、17 d,摘除幼鼠眼球作石蜡切片并行苏木精-伊红(HE)和脱氧核苷酸末端转移酶介导缺口末端标记(TUNEL)染色.观察视网膜无灌注区形态学变化,检测视网膜神经细胞凋亡数量.结果 正常对照组幼鼠视网膜各层结构清晰.出生后12 d,OIR对照组幼鼠视网膜内核层(INL)可见少量染色质凝聚的细胞核.出生后14 d,OIR对照组幼鼠视网膜INL可见大量染色质凝聚的细胞核,OIR药物组幼鼠视网膜INL均可见少量染色质凝聚的细胞核.出生后17 d,OIR对照组幼鼠视网膜INL、内丛状层(IPL)和外丛状层(OPL)厚度变薄;OIR药物组幼鼠视网膜中周部INL、IPL和OPL厚度较正常对照组薄,较OIR对照组厚.出生后12 d,OIR对照组幼鼠视网膜TUNEL阳性细胞数量为正常对照组的6倍,差异有统计学意义(t=9.432,P＜0.001).出生后14 d,3组幼鼠视网膜TUNEL阳性细胞数量比较,差异有统计学意义(F=587.217,P＜0.001);OIR对照组幼鼠视网膜TUNEL阳性细胞数量为正常对照组的28倍,差异有统计学意义(t=49.813,P＜0.001);OIR药物组幼鼠视网膜TUNEL阳性细胞数量较OIR对照组减少了82.3%,差异有统计学意义(t=42.434,P＜0.001).出生后17 d,3组幼鼠视网膜TUNEL阳性细胞数量比较,差异无统计学意义(F=587.217,P>0.05).结论 TMP能明显抑制OIR中视网膜神经细胞凋亡.

abstractsObjective To observe the inhibitory effect of tetramethylpyrazine(TMP)on apoptosis of retinal neurons in oxygen-induced retinopathy(OIR).Methods 48 C57BL/6 mice were randomly divided into normal control group(n=18),OIR control group(n=18)and OIR TMP group(n=12).The mice of normal control group were raised in room air.From the postnatal day 7(P7),mice of the other two groups mice of OIR TMP group received intraperitoneal injection of TMP(200 mg/kg)once a day from P12 to P16.meanwhile the mice of normal control group and OIR control group were iniected with the same volume of normal saline containing of 0.1% DMSO.At P12,P14 and P17,the morphologic changes in retinal avascular zone and the number of retinal apoptotic cell were observed by HE staining and TUNEL assay.Results At P1 2.there were a few of chromatin condensation and pycnic nuclei in the inner nuclear layer (INL)of OIR control group.At P14,a great quantity(OIR control group)or some(OID TMP treated group)chromatin condensation and pycnic nuclei in the central INL were observed.At P17,the thickness of INL,inner plexiform layer(IPL)and outer plexiform layer(OPL)in the OIR control group were reduced;the thickness of INL,IPL and OPL in the OIR TMP group weas thinner than those in the normal control group and thicker than those in the OIR control group.At P12,the TUNEL-positive cells in the OIR control group was 6 times of the normal eontrol group(F=587.217,P＜0.001).At P14,the difference of TUNEL-positive cells in three groups was significant(F=587.217,P＜0.001);the TUNEL-positive cells in the OIR control group was 28 times of the normal control group(t=49.813,P＜0.001);the TUNEL-positive cells in the OIR TMP group has reduced 50% compared with the OIR controI group(t=42.434,P＜0.00 1).At P17,there was no significant difference in TUNEL-positive cells among the three groups (F=587.217,P>0.05).Conclusions TMP can inhibit apoptosis of retinal cells in OIR significantly.