论文部分内容阅读
目的评价巯螺旋体素(Borrelidin)对小鼠脾窦内皮细胞凋亡的诱导作用,为静脉畸形的研究和治疗探索新的药物。方法 180只10周龄雄性SPF小鼠(specific pathogen free muose,无特定病原体小鼠),质量25-30g随机分为3组:空白对照组、生理盐水组、Borrelidin组,每组60只。每组又随机分四个日程组(3、7、14、21d组),每组15只。小鼠脾脏直接注射生理盐水、Borrelidin后分别在3、7、14、21d四个时间点处死该组小鼠取其脾脏为标本:①石蜡切片光镜观察;②超薄切片电镜观察;③末端转移酶介导的缺口末端标记法(TUNEL法)标记凋亡细胞;④免疫组化(SABC法)检测细胞凋亡因子半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)的表达;⑤图像分析技术比较Borrelidin不同时段的凋亡率。结果空白对照组和盐水组未发现脾脏萎缩和脾窦组织结构破坏。Borrelidin3天组脾脏有不同程度肿胀,7天肿胀基本消失,14天有不同程度萎缩,21天有肉眼可见的明显萎缩;电镜显示Borrelidin组各时段均可见凋亡细胞,随药物作用时间延长凋亡细胞逐渐增多;TUNEL标记显示Borrelidin作用后3、7天组就有阳性表达,14天后凋亡广泛表达于脾窦内皮细胞;免疫组化显示Caspase-3的表达与凋亡的阳性表达趋势一致。显微图像及统计学分析Borrelidin组凋亡指数及Caspase-3的表达阳性率高,其统计学差异显著。结论 Borrelidin可通过Caspase-3途径诱导小鼠脾窦内皮细胞凋亡,可用于对静脉畸形治疗的进一步研究。
Objective To evaluate the induction of Borrelidin on the apoptosis of mouse splenic sinus endothelial cells and to explore new drugs for the study and treatment of venous malformations. Methods 180 male SPF mice (specific pathogen free muose) of 10 weeks old were randomly divided into 3 groups: blank control group, saline group and Borrelidin group, with 60 mice in each group. Each group was randomly divided into four calendars (3,7,14,21d group), each group of 15. The spleen of mice were directly injected with normal saline and Borrelidin and the spleens were sacrificed at 3, 7, 14, and 21 days respectively. The spleens were taken from the spleen of the mice for three days: ① paraffin sections were observed by light microscopy; ② ultrathin section electron microscopy; The apoptotic cells were labeled by TUNEL method; ④ The expression of caspase-3 was detected by immunohistochemistry (SABC method) ⑤ image analysis of Borrelidin at different stages of apoptosis rate. Results No spleen atrophy and splenic tissue structure were found in blank control group and saline group. Borrelidin 3 days group had different degrees of swelling of the spleen, 7 days swelling basically disappeared, varying degrees of 14 days of atrophy, 21 days visible macroscopic atrophy; electron microscopy showed Borrelidin group of apoptotic cells were observed in each time period, with the drug effect of prolonged apoptosis TUNEL staining showed that Borrelidin had a positive expression on days 3 and 7, and after 14 days, apoptosis was widely expressed in splenic sinus endothelial cells. Immunohistochemistry showed that the expression of Caspase-3 was consistent with the positive expression of apoptosis. Microscopic image and statistical analysis Borrelidin group apoptosis index and Caspase-3 expression positive rate, the statistical difference was significant. Conclusion Borrelidin can induce the apoptosis of splenic sinusoidal endothelial cells in mice via Caspase-3 pathway and can be used for the further study of treatment of venous malformations.