不同高压氧吸氧时间对脑出血大鼠血肿周围B淋巴细胞瘤-2基因和BCL2-Associated X蛋白表达的影响

Hyperbaric oxygen inhalation downregulates the expression of BCL2-associated protein X and upregulates the expression of the B lymphoblastoma-2 gene after cerebral hemorrhage

目的观察不同高压氧(HBO)吸氧时间对脑出血(ICH)大鼠血肿周围B淋巴细胞瘤-2基因(Bcl-2)和BCL2-Associated X蛋白(Bax)表达的影响,以期探索HBO治疗脑出血的治疗机制和最佳吸氧时间。方法选取Wistar大鼠130只,其中正常大鼠5只(正常组),造模大鼠125只(胶原酶诱导法建立ICH模型),按随机数字表法将造模大鼠随机分为对照组(无高压氧干预)25只和实验组(高压氧干预)100只,实验组根据吸氧时间的不同再按随机数字表法分为HBO 40 min组,HBO 60 min组,HBO 80 min组,HBO 100 min组,共4个亚组,每组25只大鼠。4个实验组亚组大鼠分别于ICH后6 h后行对应的HBO治疗,每日治疗1次,于治疗第1、3、5、7、14天后每个时间点各处死组内5只大鼠(对照组于相同时间点各处死组内5只大鼠,正常组于入组当天处死并取材)。采用免疫组化法检测各组大鼠各取材时间点血肿周边脑组织中Bax和Bcl-2蛋白的表达情况。结果HBO 40 min组、HBO 60 min组、HBO 80 min组、HBO 100 min组各时间点的Bax表达较对照组相同时间点均显著减少,差异均有统计学意义(P<0.05)。其中HBO 60 min组和HBO 80 min组的Bax表达分别与HBO40 min组和HBO 100 min组相同时间点比较,差异均有统计学意义(P<0.05)。造模成功3 d、5 d、7 d、14 d后,HBO 40 min组和HBO 60 min组的Bcl-2表达分别与HBO 80 min组和HBO 100 min组同时间点比较,差异均有统计学意义(P<0.05)。HBO 80 min组在造模成功7 d和14 d后的Bcl-2/Bax比值分别为1.5792和1.5733,与HBO 40 min组、HBO 60 min组以及HBO 100 min组比较,差异均有统计学意义(P<0.05)。结论HBO治疗ICH的机制可能与其可以下调出血灶周围Bax的表达水平以及提高Bcl-2的表达水平有关,而HBO治疗ICH的最佳吸氧时间是80 min。

Objective To explore any effect of inhaling hyperbaric oxygen on edema resulting from cerebral hemorrhage.Methods Cerebral hemorrhage was modeled using collagenase in 125 Wistar rats,with 5 others used as controls.The injured rats were randomly divided into a second control group without any hyperbaric oxygen inter-vention,and groups which inhaled hyperbaric oxygen for 40 min,60 min,80 min or 100 min.There were 25 rats in each group.The inhalation began 6 hours after the modeling and was administered once a day.On the 2nd,4th,6th,8th and 16th day of the treatment,5 rats from each group were sacrificed and the expression of the B lymphoblastoma-2 gene(Bcl-2)and BCL2-associated protein X(Bax)in the brain tissue around the hematoma was detected using immunohistochemical methods.Results The average Bax expression in all of the HBO groups was significantly lower than in the second control group at each time point,but with significant differences between the HBO 60 min and HBO 40 min groups,as well as between the HBO 80 min and HBO 100 min groups.By the 4th day the average Bcl-2 expression in the HBO 40 min group was significantly different from that in the HBO 80 min group,and the same was true of the HBO 60 min and HBO 100 min groups.Conclusions One of the mechanisms by which HBO treats intracerebral hemorrhage may be related to down-regulating the expression of Bax and increasing the expression of Bcl-2.The best everyday treatment duration is 80 min.