两种方法清除大鼠子宫内膜巨噬细胞的比较研究OA北大核心CSTPCD
A comparative study of two methods for the clearance of macrophages from rat endometrium
目的 通过比较氯膦酸二钠脂质体不同给药途径的作用差异,探索更有效的大鼠子宫内膜巨噬细胞清除方法.方法 雌性8周龄SD大鼠随机分为单侧对照组(左侧宫腔注射磷酸缓冲盐脂质体100 μL)、单侧清除组(右侧宫腔注射氯膦酸二钠脂质体100 μL)、双侧对照组(双侧宫腔注射磷酸缓冲盐脂质体100μL)、双侧清除组(双侧宫腔注射氯膦酸二钠脂质体100 μL)、全身对照组(尾静脉注射磷酸缓冲盐脂质体500 μL)和全身清除组(尾静脉注射氯膦酸二钠脂质体500 μL).注射后48 h,获取组织样本.采用HE染色观察子宫、卵巢组织形态结构,免疫组化观察子宫、卵巢组织中巨噬细胞阳性表达,流式细胞术检测子宫、卵巢组织中巨噬细胞数量的变化.结果 各组大鼠子宫、卵巢组织结构与形态并无明显差异.免疫组化法结果显示,与对照组相比,单侧及双侧清除组中子宫巨噬细胞阳性表达明显降低(P<0.001),卵巢巨噬细胞阳性表达无影响(P>0.05),全身清除组子宫和卵巢组织中巨噬细胞阳性表达均减少(P<0.01,P<0.01);与单侧及双侧清除组相比,全身清除对卵巢组织中巨噬细胞阳性表达的影响更大(P<0.05).流式细胞术检测显示,与对照组相比,各清除组均能有效降低子宫组织中巨噬细胞百分比(P<0.001,P<0.001,P<0.05),与全身清除组相比,单侧及双侧清除组子宫内膜巨噬细胞清除效果更好(P<0.05,P<0.05);各清除组卵巢组织中巨噬细胞数量与对照组相比均有所下降,其中全身清除组巨噬细胞数目减少最明显(P<0.05),单侧及双侧清除组与对照组相比差异无统计学意义(P>0.05).结论 局部宫腔注射氯膦酸二钠脂质体清除子宫巨噬细胞的方式优于尾静脉注射,并可避免其对卵巢巨噬细胞的影响,是建立理想的子宫局部巨噬细胞清除模型的最佳方法.
Objective A comparison of different routes for the administration of clodronate disodium liposomes to determine the most effective method of rat endometrial macrophage clearance.Methods Female 8-week-old SD rats were randomly divided into a unilateral control group(injected with 100 μL PBS liposomes into the left uterine cavity),unilateral clearance group(injected with 100 μL clodronate liposomes into the right uterine cavity),bilateral control group(injected with 100 μL PBS liposomes into the bilateral intrauterine),bilateral clearance group(injected with 100 μL clodronate liposomes into the bilateral intrauterine),whole-body control group(injected with 500 μL PBS liposomes into the caudal vein),and whole-body clearance group(injected with 500 μL clodronate liposomes into the caudal vein).Hematoxylin and eosin staining was used to observe the morphology and structures of uterine and ovarian tissues,immunohistochemistry was used to observe the presence of macrophages in uterine and ovarian tissues,and flow cytometry was used to detect changes to macrophage cell counts in uterine and ovarian tissues.Results There were no significant differences in the structures or morphology of the uterus and ovary among the groups.Immunohistochemical staining showed that,compared to the control group,the unilateral and bilateral uterine clearance groups'population of terine macrophages was significantly decreased(P<0.001),but there was no difference in the accumulation of macrophages in the ovary(P>0.05).The number of macrophages in both uterine and ovarian tissues decreased in the whole-body clearance group(P<0.01,P<0.01).Compared with the unilateral and bilateral clearance groups,the whole-body clearance group had more macrophages in the ovarian tissues(P<0.05).Flow cytometry showed that,compared with the control group,each clearance group's percentages of macrophages in the uterine tissue were significantly reduced(P<0.001,P<0.001,P<0.05).Compared to the whole-body clearance group,the unilateral and bilateral clearance groups'endometrial macrophages had superior clearance activity(P<0.05,P<0.05).In addition,the number of macrophages in ovarian tissue decreased in all clearance groups compared to the control group.The decrease in macrophage number was most pronounced in the whole-body clearance group(P<0.05),and there was no significant difference in numbers between the unilateral and bilateral clearance groups and the control group(P>0.05).Conclusions Local injection of clodronate liposomes was more effective than caudal injection for clearing uterine macrophages,and the impact on ovarian macrophages was largely avoided.Thus local clodronate liposome injection is an improved method for establishing a local uterine macrophage clearance model.
夏良君;崔础婷;李军威;程洁;夏有兵
南京中医药大学针灸推拿学院·养生康复学院,南京 210023南京中医药大学针灸推拿学院·养生康复学院,南京 210023||南京医科大学,南京 211166
子宫内膜巨噬细胞宫腔注射氯膦酸二钠脂质体
endometrial macrophagesintrauterine injectionclodronate liposome
《中国比较医学杂志》 2024 (004)
84-91 / 8
国家自然科学基金(82205251,82274638);江苏省高校自然科学研究重大项目(23KJA360006);国家自然科学基金校配套项目(XPT82205251).
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