中华内科杂志
中華內科雜誌
중화내과잡지
CHINESE JOURNAL OF INTERNAL MEDICINE
2012年
10期
793-797
,共5页
伍丽萍%施秉银%旬利茹%郭立英%杨靖%徐利
伍麗萍%施秉銀%旬利茹%郭立英%楊靖%徐利
오려평%시병은%순리여%곽립영%양정%서리
Graves病%小鼠%模型,动物%诱导
Graves病%小鼠%模型,動物%誘導
Graves병%소서%모형,동물%유도
Graves disease%Mice%Models,animal%Induce
目的 对比促甲状腺素受体(TSHR)质粒DNA(pcDNA3.1-TSHR)及TSHR A亚单位重组腺病毒(Ad-TSHR289)诱导Graves病动物模型的效率,研究造模持续时间对Ad-TSHR289诱导的Graves病甲状腺功能亢进(甲亢)及相关指标的影响.方法 分别设质粒组和病毒组.质粒组:将21只雌性BALB/c小鼠随机分为造模组(12只)、对照组(9只),造模组给予皮内注射pcDNA3.1-TSHR 50 μg,每3周1次,共免疫3次,末次免疫后4周处死小鼠,取血测TSHR抗体(TRAb)、总T4,剥离甲状腺行组织学检查.对照组给予等剂量的pcDNA3.1注射.病毒组:将52只雌性BALB/c小鼠分为10周造模组(8只)、14周造模组(10只)、18周造模组(8只)并分别设对照组(依次为8只/组、10只/组、8只/组),造模组给予Ad-TSHR289肌肉注射,每3周1次,共3次,末次免疫后4周、8周、12周依次处死小鼠,测定血清TRAb、总T4水平并观察甲状腺组织病理变化.对照组给予等剂量的Ad-lacz以相同方式处理.另设8只小鼠分别于3次免疫前后检测TRAb动态变化.结果 质粒造模组只有2只小鼠出现较弱的TSHR抗体反应,无总T4水平的升高及甲状腺组织增生性改变.病毒10周造模组所有小鼠均出现了高水平的TRAb[(807.65±136.33)U/L]、8只小鼠中有6只小鼠发生甲亢并伴有甲状腺组织增生性改变.14周造模组TRAb水平[(650.12±192.88)U/L]、甲亢发生率(3/10)低于10周造模组,甲状腺增生性改变程度有所减轻,但其阳性率无降低.18周造模组8只小鼠中只有2只TRAb滴度出现轻度的升高,没有小鼠总T4水平高于正常,2只小鼠甲状腺组织轻度增生.进行TRAb滴度动态观察的小鼠抗体水平和不同时间的造模组呈现相似趋势.结论 Ad-TSHR289诱导的Graves病动物模型重复性好,甲亢发生率高,仍是目前较为理想的研究工具.该模型持续时间可达18周,而10周为维持Graves病疾病特征最佳时段.
目的 對比促甲狀腺素受體(TSHR)質粒DNA(pcDNA3.1-TSHR)及TSHR A亞單位重組腺病毒(Ad-TSHR289)誘導Graves病動物模型的效率,研究造模持續時間對Ad-TSHR289誘導的Graves病甲狀腺功能亢進(甲亢)及相關指標的影響.方法 分彆設質粒組和病毒組.質粒組:將21隻雌性BALB/c小鼠隨機分為造模組(12隻)、對照組(9隻),造模組給予皮內註射pcDNA3.1-TSHR 50 μg,每3週1次,共免疫3次,末次免疫後4週處死小鼠,取血測TSHR抗體(TRAb)、總T4,剝離甲狀腺行組織學檢查.對照組給予等劑量的pcDNA3.1註射.病毒組:將52隻雌性BALB/c小鼠分為10週造模組(8隻)、14週造模組(10隻)、18週造模組(8隻)併分彆設對照組(依次為8隻/組、10隻/組、8隻/組),造模組給予Ad-TSHR289肌肉註射,每3週1次,共3次,末次免疫後4週、8週、12週依次處死小鼠,測定血清TRAb、總T4水平併觀察甲狀腺組織病理變化.對照組給予等劑量的Ad-lacz以相同方式處理.另設8隻小鼠分彆于3次免疫前後檢測TRAb動態變化.結果 質粒造模組隻有2隻小鼠齣現較弱的TSHR抗體反應,無總T4水平的升高及甲狀腺組織增生性改變.病毒10週造模組所有小鼠均齣現瞭高水平的TRAb[(807.65±136.33)U/L]、8隻小鼠中有6隻小鼠髮生甲亢併伴有甲狀腺組織增生性改變.14週造模組TRAb水平[(650.12±192.88)U/L]、甲亢髮生率(3/10)低于10週造模組,甲狀腺增生性改變程度有所減輕,但其暘性率無降低.18週造模組8隻小鼠中隻有2隻TRAb滴度齣現輕度的升高,沒有小鼠總T4水平高于正常,2隻小鼠甲狀腺組織輕度增生.進行TRAb滴度動態觀察的小鼠抗體水平和不同時間的造模組呈現相似趨勢.結論 Ad-TSHR289誘導的Graves病動物模型重複性好,甲亢髮生率高,仍是目前較為理想的研究工具.該模型持續時間可達18週,而10週為維持Graves病疾病特徵最佳時段.
목적 대비촉갑상선소수체(TSHR)질립DNA(pcDNA3.1-TSHR)급TSHR A아단위중조선병독(Ad-TSHR289)유도Graves병동물모형적효솔,연구조모지속시간대Ad-TSHR289유도적Graves병갑상선공능항진(갑항)급상관지표적영향.방법 분별설질립조화병독조.질립조:장21지자성BALB/c소서수궤분위조모조(12지)、대조조(9지),조모조급여피내주사pcDNA3.1-TSHR 50 μg,매3주1차,공면역3차,말차면역후4주처사소서,취혈측TSHR항체(TRAb)、총T4,박리갑상선행조직학검사.대조조급여등제량적pcDNA3.1주사.병독조:장52지자성BALB/c소서분위10주조모조(8지)、14주조모조(10지)、18주조모조(8지)병분별설대조조(의차위8지/조、10지/조、8지/조),조모조급여Ad-TSHR289기육주사,매3주1차,공3차,말차면역후4주、8주、12주의차처사소서,측정혈청TRAb、총T4수평병관찰갑상선조직병리변화.대조조급여등제량적Ad-lacz이상동방식처리.령설8지소서분별우3차면역전후검측TRAb동태변화.결과 질립조모조지유2지소서출현교약적TSHR항체반응,무총T4수평적승고급갑상선조직증생성개변.병독10주조모조소유소서균출현료고수평적TRAb[(807.65±136.33)U/L]、8지소서중유6지소서발생갑항병반유갑상선조직증생성개변.14주조모조TRAb수평[(650.12±192.88)U/L]、갑항발생솔(3/10)저우10주조모조,갑상선증생성개변정도유소감경,단기양성솔무강저.18주조모조8지소서중지유2지TRAb적도출현경도적승고,몰유소서총T4수평고우정상,2지소서갑상선조직경도증생.진행TRAb적도동태관찰적소서항체수평화불동시간적조모조정현상사추세.결론 Ad-TSHR289유도적Graves병동물모형중복성호,갑항발생솔고,잉시목전교위이상적연구공구.해모형지속시간가체18주,이10주위유지Graves병질병특정최가시단.
Objective To compare the efficacy of Graves disease animal models induced by thyroid stimulating hormone receptor (TSHR) plasmid DNA (pcDNA3.1-TSHR) and by TSHR A subunit recombinant adenovirus(Ad-TSHR289),and to investigate the influence of duration for preparing animal model induced by Ad-TSHR289 on Graves hyperthyroidism and its related indices.Methods The plasmid group and the adenovirus group were set up respectively.The plasmid group:21 female BALB/c mice were randomly divided into model group (n =12) and control group (n =9).The model group were injected intradermally with pcDNA3.1-TSHR 50 μg,once every 3 weeks,totally 3 times.Then 4 weeks after the last immunization,the mice were euthanized to obtain blood for testing TSHR antibody (TRAb),total T4,and thyroid tissue for histological examination.The controls were injected with the same dose of pcDNA3.1 in the same way.The adenovirus group:52 female BALB/c mice were divided into 10-week model group (n =8),14-week model group (n =10) and 18-week model group (n =8),and the respective controls (n =8,n =10,n =8) were set up.All model groups were injected intramuscularly with Ad-TSHR289,three times at three weekly intervals.Then the mice were euthanized at 4,8 and 12 weeks to test TRAb,total T4 level and to observe the change of thyroid histology.The controls were treated with the same dose of Ad-lacz in the same way.Another 8 mice were scheduled to test the dynamic variation of TRAb before and after the 3 times immunization.Results In the plasmid model group,only two of 12 mice developed weak antibody responses against TSHR,and no elevated total T4 level and no hyperplasia changes of thyroid were observed.In the 10-week model group,all mice had high level TRAb [(807.65 ± 136.33)U/L,Six-eighths mice had hyperthyroidism exhibited hyperplasia changes.In the 14-week model group,the TRAb level [(650.12 ± 192.88) U/L]and the incidence of hyperthyroidism (3/10) were lower than those in 10-week group.Histologically,the degree of thyroid hyperplasia lightened to a small extent,but its positive rate did not decline.In the 18-week model group,only 2 of 8 mice displayed slightly elevated TRAb level,and no mice showed increased total T4 level.Additionally,thyroid tissues of 2 mice were mildly abnormal.Compared with the model groups at different time,the change of antibody levels of the mice for TRAb dynamic observation exhibited the similar trend.Conclusions Being good at repeatability and high incidence of hyperthyroidism,the animal model of Graves disease induced by Ad-TSHR289 is still an ideal research tool presently.The duration of model ean be maintained 18 weeks,and 10 weeks is the best period to snstain characteristic of Graves disease.
