目的:探究鸢尾素对新生大鼠高胆红素血症细胞凋亡的影响.方法:将96只新生SD大鼠随机分为对照组(N组)和模型组(M组),于7和10日龄腹腔注射胆红素溶液进行造模.造模成功后将M组随机分为T0组、T1组、T2组、T3组.T1、T2、T3组分别侧脑室注射40、60和80 μg/kg的鸢尾素溶液,余各组注射等量磷酸盐缓冲液溶液.侧脑室注射24h后,神经行为学、苏木精-伊红染色(Hematoxylin-Eosin staining,HE)、原位末端标记法(terminal-deoxynucleoitidyl transferase mediated nick end labeling,Tunel)检测海马区病理改变并统计凋亡率,Western Blot检测海马Bcl-2、Bax和Caspase-3蛋白相对表达量.结果:侧脑室注射鸢尾素后.平面翻正反射:T3组所需时间少于T0、T1、T2组且多于N组(P＜0.01);负趋地性反射:T3组所需时间少于T0、T1、T2组(P＜0.01,P＜0.05).HE染色:M组海马神经细胞核固缩、裂解,排列紊乱.T1、T2、T3组细胞形态等不同程度改善.Tunel染色:M组Tunel+细胞增多,布满视野;N组Tunel+细胞少,呈零星分布;T3组凋亡率低于T0、T1组(P＜0.05).Western Blot:各组Bcl-2蛋白相对表达量无显著性意义(P＞0.05);T0组Bax蛋白高于其余各组(P＜0.05),T3组低于T1组(P＜0.05);T0组Bcl-2/Bax值低于N、T3组(P＜0.05),T1、T2组均低于T3组(P＜0.05);T0组Caspase-3蛋白高于其余各组(P＜0.05),N组低于T1组(P＜0.05),T1组高于T3组(P＜0.05).结论:鸢尾素可能通过减少Bax、Caspase-3蛋白表达,升高Bcl-2/Bax值来减轻神经细胞凋亡且与鸢尾素注射剂量相关.
Effects of irisin on apoptosis in neonatal rats with hyperbilirubinemia
Objective:To investigate the effect of irisin on apoptosis of hyperbilirubinemia cells in neonatal rats.Method:Ninety-six newbom SD rats were randomly divided into the control group(N group)and the model group(M group).The model was established by intraperitoneal injection of bilirubin solution at age 7 and 10 days.After successful modeling,the M group was randomly divided into four groups:T0,T1,T2,T3.Groups T1,T2,and T3 received intracerebroventricular injections of 40,60,and 80 μg/kg irisin solution,re-spectively,while the other groups were injected with an equivalent volume of phosphate buffer saline(PBS)solution.After 24 hours,neurobehavioral assessments were conducted,hematoxylin-eosin(HE)staining,termi-nal-deoxynucleoitidyl transferase mediated Nick end labeling(Tunel)were used to detect the hippocampus pathological changes and apoptosis rate statistics,and Westem Blot was used to detect relative expression lev-els of Bcl-2,Bax and Caspase-3 proteins in hippocampus.Result:After irisin injection,the T3 group required less time needed for planar double positive reflection than the T0,T1,T2 and more than N group(P＜0.01);negative geotaxis reflex time was also less in T3 group compared to T0,T1,T2 groups(P＜0.01,P＜0.05).HE staining showed nuclear pyknosis,cleavage,and disor-dered arrangement of hippocampal neurons in the M group.The cell morphology was improved to varying de-grees in T1,T2 and T3 groups.Tunel staining showed increased Tunel+cells throughout the field in M group,sparse distribution in N group and lower apoptosis rates in T3 group compared to T0 and T1 groups(P＜0.05).Western Blot showed no significant difference in the relative expression of Bcl-2 protein among all groups(P＞0.05);T0 group had higher Bax protein levels compared to the rest of groups(P＜0.05),and lower in T3 group compared to T1 group(P＜0.05);The Bcl-2/Bax ratio was lower in T0 group compared to N and T3 group(P＜0.05),and lower in T1,T2 groups compared to T3 group(P＜0.05);The expression of Cas-pase-3 protein were higher in T0 group than that in other groups(P＜0.05),lower in N group than T1 group(P＜0.05),and higher in T1 group than T3 group(P＜0.05).Conclusion:Irisin may reduce the apoptosis of nerve cells by reducing the expression of Bax and Caspase-3 protein and increasing the Bcl-2/Bax ratio,with effects dependent on the injection dose of irisin.
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