目的 观察碘缺乏和甲状腺功能减退(甲减)对大鼠仔鼠小脑细胞外信号调节激酶1/2(extracellular signal-regulate kinase,ERK1/2)蛋白表达的影响.方法 健康Wistar大鼠28只,雌性,60日龄.将大鼠按体质量随机分为对照组、碘缺乏病组、甲减组,甲减组根据饮水中含丙基硫尿嘧啶(PTU)剂量分为5 mg/L组和15 mg/L组.每组各7只.母鼠妊娠后,分别选用低碘饲料及PTU饮水诱导建立低碘及甲减大鼠动物模型.仔鼠出生7、14、21、28、42 d时分别处死,测定仔鼠小脑质量;取小脑组织进行镀银染色和免疫组织化学链霉菌抗生物素蛋白-过氧化物酶(S-P)染色,光镜下观察小脑形态学变化,图像分析仪下观察ERK1/2蛋白表达.结果 仔鼠出生14、21、28、42 d时,小脑质量各组间比较差异有统计学意义(F=6.325、8.870、16.191、21.574,P均<0.05);碘缺乏病组仔鼠小脑质量[(0.0945±0.0233)、(0.1347±0.0046)、(0.1542±0.0094)、(0.1949±0.0048)g]明显低于对照组[(0.1856±0.0123)、(0.2049±0.0098)、(0.2268±0.0065)、(0.2606±0.0086)g,P均<0.05]和甲减组[5 mg/L组:(0.1741±0.0172)、(0.1927±0.0103)、(0.2181±0.0064)、(0.2583±0.0054)g,P均<0.05;15 mg/L组:(0.1604±0.0083)、(0.1682±0.0103)、(0.1996±0.0073)、(0.2579±0.0067)g,P均<0.05].出生7 d时,对照组、甲减组仔鼠小脑皮质具有典型的层状结构,各层之间界限清晰,但碘缺乏病组各层之间界限模糊;出生21 d时,与对照组比较,15 mg/L组和碘缺乏病组仔鼠小脑外颗粒细胞消失延迟,仍有2～3层外颗粒细胞;出生28、42 d时,甲减组和碘缺乏病组仔鼠小脑分子层厚度低于对照组.小脑组织ERK1/2平均积分光密度,在出生7 d时,组间比较差异无统计学意义(F=1.102,P>0.05);在出生14、21、28、42 d时,各组间比较差异有统计学意义(F=16.131、13.543、26.953、41.583,P均<0.01);碘缺乏病组(7.3245±0.5070)、(8.3606±1.0683)、(9.1217±1.0402)、(12.1587±0.7581)和甲减组[5 mg/L组:(11.4307±15200)、(14.919±0.8497)、(169082±1.1130)、(15.7721±0.82913);15 ms/L组:(7.8538±0.9775)、(11.2461±0.8138)、(12.7800±1.3783)、(13.0871±1.1450)]明显低于对照组[(16.2831±0.5143)、(20.2653±0.9551)、(22.7485±1.0267)、(22.1725±0.9939),P均<0.01].结论 碘缺乏和甲减可使仔鼠小脑产生明显的形态学改变,降低大鼠仔鼠小脑组织ERK1/2蛋白表达.影响神经系统的发育.
目的 觀察碘缺乏和甲狀腺功能減退(甲減)對大鼠仔鼠小腦細胞外信號調節激酶1/2(extracellular signal-regulate kinase,ERK1/2)蛋白錶達的影響.方法 健康Wistar大鼠28隻,雌性,60日齡.將大鼠按體質量隨機分為對照組、碘缺乏病組、甲減組,甲減組根據飲水中含丙基硫尿嘧啶(PTU)劑量分為5 mg/L組和15 mg/L組.每組各7隻.母鼠妊娠後,分彆選用低碘飼料及PTU飲水誘導建立低碘及甲減大鼠動物模型.仔鼠齣生7、14、21、28、42 d時分彆處死,測定仔鼠小腦質量;取小腦組織進行鍍銀染色和免疫組織化學鏈黴菌抗生物素蛋白-過氧化物酶(S-P)染色,光鏡下觀察小腦形態學變化,圖像分析儀下觀察ERK1/2蛋白錶達.結果 仔鼠齣生14、21、28、42 d時,小腦質量各組間比較差異有統計學意義(F=6.325、8.870、16.191、21.574,P均<0.05);碘缺乏病組仔鼠小腦質量[(0.0945±0.0233)、(0.1347±0.0046)、(0.1542±0.0094)、(0.1949±0.0048)g]明顯低于對照組[(0.1856±0.0123)、(0.2049±0.0098)、(0.2268±0.0065)、(0.2606±0.0086)g,P均<0.05]和甲減組[5 mg/L組:(0.1741±0.0172)、(0.1927±0.0103)、(0.2181±0.0064)、(0.2583±0.0054)g,P均<0.05;15 mg/L組:(0.1604±0.0083)、(0.1682±0.0103)、(0.1996±0.0073)、(0.2579±0.0067)g,P均<0.05].齣生7 d時,對照組、甲減組仔鼠小腦皮質具有典型的層狀結構,各層之間界限清晰,但碘缺乏病組各層之間界限模糊;齣生21 d時,與對照組比較,15 mg/L組和碘缺乏病組仔鼠小腦外顆粒細胞消失延遲,仍有2～3層外顆粒細胞;齣生28、42 d時,甲減組和碘缺乏病組仔鼠小腦分子層厚度低于對照組.小腦組織ERK1/2平均積分光密度,在齣生7 d時,組間比較差異無統計學意義(F=1.102,P>0.05);在齣生14、21、28、42 d時,各組間比較差異有統計學意義(F=16.131、13.543、26.953、41.583,P均<0.01);碘缺乏病組(7.3245±0.5070)、(8.3606±1.0683)、(9.1217±1.0402)、(12.1587±0.7581)和甲減組[5 mg/L組:(11.4307±15200)、(14.919±0.8497)、(169082±1.1130)、(15.7721±0.82913);15 ms/L組:(7.8538±0.9775)、(11.2461±0.8138)、(12.7800±1.3783)、(13.0871±1.1450)]明顯低于對照組[(16.2831±0.5143)、(20.2653±0.9551)、(22.7485±1.0267)、(22.1725±0.9939),P均<0.01].結論 碘缺乏和甲減可使仔鼠小腦產生明顯的形態學改變,降低大鼠仔鼠小腦組織ERK1/2蛋白錶達.影響神經繫統的髮育.
목적 관찰전결핍화갑상선공능감퇴(갑감)대대서자서소뇌세포외신호조절격매1/2(extracellular signal-regulate kinase,ERK1/2)단백표체적영향.방법 건강Wistar대서28지,자성,60일령.장대서안체질량수궤분위대조조、전결핍병조、갑감조,갑감조근거음수중함병기류뇨밀정(PTU)제량분위5 mg/L조화15 mg/L조.매조각7지.모서임신후,분별선용저전사료급PTU음수유도건립저전급갑감대서동물모형.자서출생7、14、21、28、42 d시분별처사,측정자서소뇌질량;취소뇌조직진행도은염색화면역조직화학련매균항생물소단백-과양화물매(S-P)염색,광경하관찰소뇌형태학변화,도상분석의하관찰ERK1/2단백표체.결과 자서출생14、21、28、42 d시,소뇌질량각조간비교차이유통계학의의(F=6.325、8.870、16.191、21.574,P균<0.05);전결핍병조자서소뇌질량[(0.0945±0.0233)、(0.1347±0.0046)、(0.1542±0.0094)、(0.1949±0.0048)g]명현저우대조조[(0.1856±0.0123)、(0.2049±0.0098)、(0.2268±0.0065)、(0.2606±0.0086)g,P균<0.05]화갑감조[5 mg/L조:(0.1741±0.0172)、(0.1927±0.0103)、(0.2181±0.0064)、(0.2583±0.0054)g,P균<0.05;15 mg/L조:(0.1604±0.0083)、(0.1682±0.0103)、(0.1996±0.0073)、(0.2579±0.0067)g,P균<0.05].출생7 d시,대조조、갑감조자서소뇌피질구유전형적층상결구,각층지간계한청석,단전결핍병조각층지간계한모호;출생21 d시,여대조조비교,15 mg/L조화전결핍병조자서소뇌외과립세포소실연지,잉유2～3층외과립세포;출생28、42 d시,갑감조화전결핍병조자서소뇌분자층후도저우대조조.소뇌조직ERK1/2평균적분광밀도,재출생7 d시,조간비교차이무통계학의의(F=1.102,P>0.05);재출생14、21、28、42 d시,각조간비교차이유통계학의의(F=16.131、13.543、26.953、41.583,P균<0.01);전결핍병조(7.3245±0.5070)、(8.3606±1.0683)、(9.1217±1.0402)、(12.1587±0.7581)화갑감조[5 mg/L조:(11.4307±15200)、(14.919±0.8497)、(169082±1.1130)、(15.7721±0.82913);15 ms/L조:(7.8538±0.9775)、(11.2461±0.8138)、(12.7800±1.3783)、(13.0871±1.1450)]명현저우대조조[(16.2831±0.5143)、(20.2653±0.9551)、(22.7485±1.0267)、(22.1725±0.9939),P균<0.01].결론 전결핍화갑감가사자서소뇌산생명현적형태학개변,강저대서자서소뇌조직ERK1/2단백표체.영향신경계통적발육.
Objective To study the effects of iodine deficiency and hypothyroidism on protein expression of extracellular signal-regulate kinase(ERK1/2) in the cerebellum of rots. Methods Twenty-eight healthy Wistar rots, female, 60 days old, were randomly divided according to their body weight into control group, iodine deficient group and hypothyroidism groups. Hypothyroidism groups in accordance with drinking water containing propylthiouracil(PTU) were divided into doses of 5 mg/L and 15 mg/L groups, 7 rats in each group. Rats after pregnancy, iodine deficient rats were administered with iodine-deficient diet and hypothyroid rats were administered with PTU in drinking water. Pup's cerebellum in each group were weighed on day 14,21,28 and 42. Cerebellum tissue was observed for cerebellar morphology using silver staining and detected for ERK1/2 protein using immunohistochemistry on day 7,14,21,28 and 42. Results On day 14,21,28 and 42, cerebellum weight of pups from iodine-deficient[(0.0945±0.0233), (0.1347±0.0046), (0.1542±0.0094), (0.1949±0.0048)g]were significantly lighter than control[(0.1856±0.0123), (0.2049±0.0098), (0.2268±0.0065), (0.2606±0.0086)g, all P < 0.05]and hypothyroidism groups [for 5 mg/L group: (0.1741±0.0172), (0.1927±0.0103), (0.2181±0.0064), (0.2583±0.0054)g, all P<0.05; for 15 mg/L group: (0.1604±0.0083), (0.1682±0.0103), (0.1996±0.0073) and (0.2579±0.0067)g, all P< 0.05]the difference had statistical significance(F=6.325,8.870, 16.191 and 21.574, all P<0.05). Compared to the controls on day 7, iodine-deficient group didn't have clear layers; on day 21, disappearance of external granule cells from iodine-deficient and 15 mg/L groups was delayed, still two or three layer external granule cells remained; on day 28 and 42, molecular layer from 5, 15 mg/L and iodine-deficient groups and became thinner. Immunohistochemistry showed that on day 7, there was no statistical difference of integrated optical density average of ERK1/2, in all the groups(F=1.102, P>0.05); on day 14,21,28 and 42, integrated optical density average of ERK1/2 in iodine-deficient group[(7.3245±0.5070), (8.3606±1.0683), (9.1217±1.0402), (12.1587±0.7581), all P<0.01]and hypothyroidism groups [for 5 mg/L group: (11.4307±1.5200), (14.919±0.8497), (16.0082±1.1130), (15.7721±0.8293), all P< 0.01; for 15 mg/L group: (7.8538±0.9775), (11.2461±0.8138),(12.78±1.3783), (13.0871±1.1450), all P < 0.01]was significantly lower than those of controls [(16.2831±0.5143), (20.2653±0.9551), (22.7485±1.0267), (22.1725±0.9939), all P < 0.01], the difference having a statistical signifieance(F=16.131,13.543,26.953,41.583, all P<0.01). Conclusions Iodine deficiency and hypothyroidism during critical periods of brain development may change eerebellar morphology and down regulate the protein expression of ERK1/2, which may result in damage of cerebellum development.
