CAJ | 학술논문

目的观察糖尿病大鼠脑组织淀粉样前体蛋白β位分解酶1(BACE1)、β-淀粉样蛋白(Aβ)的表达,探讨糖尿病糖代谢异常在阿尔茨海默病中的作用机制.方法腹腔注射链脲佐菌素诱发糖尿病大鼠动物模型,随机分为正常组(N组)、假手术组(S组)、4周模型组(M4组)、6周模型组(M6组)、8周模型组(M8组).穿梭箱实验、Morris水迷宫实验检测大鼠认知行为学改变,免疫组织化学法、免疫印迹法、RT-PCR和酶联免疫吸附法(ELISA)检测BACE1;ELISA法检测Aβ在糖尿病大鼠脑组织中的表达,用图像分析仪测定吸光度值.结果 M组大鼠的电击次数、学习和记忆潜伏期时间显著延长(P<0.01),主动逃避次数显著降低(P<0.01);ELISA法检测Aβ_(1-40)和Aβ_(1-42)显示糖尿病模型鼠脑组织内Aβ_(1-40)水平明显增高,从正常组的(64.13±6.76)pg/mg升至模型组的(86.43±7.03)pg/mg(P<0.001);Aβ_(1-42)也明显升高,从正常组的(67.43±5.12 )pg/mg 升至(89.45±5.28) pg/mg (P<0.001);糖尿病模型鼠脑组织内BACE1水平显著增高,ELISA法从正常组的(116.46±8.10)pg/mg升至模型组的(158.73±6.24)pg/mg (P<0.001).免疫印迹法吸光度值从正常组的0.61±0.11升至模型组的1.52±0.16 (P<0.001),RT-PCR法吸光度值从正常组的1.62±0.26升至模型组的3.61±0.32 (P<0.001),免疫组织化学法吸光度值从正常组的0.81±0.21升至模型组的2.01±0.36 (P<0.001).BACE1和Aβ表达水平与学习和记忆负相关.结论 BACE1和Aβ在糖尿病大鼠脑组织表达增高,糖尿病糖代谢异常增强BACE1和Aβ表达,参与了阿尔茨海默病的发病机制.
목적 관찰당뇨병대서뇌조직정분양전체단백β위분해매1(BACE1)、β-정분양단백(Aβ)적표체,탐토당뇨병당대사이상재아이자해묵병중적작용궤제.방법 복강주사련뇨좌균소유발당뇨병대서동물모형,수궤분위정상조(N조)、가수술조(S조)、4주모형조(M4조)、6주모형조(M6조)、8주모형조(M8조).천사상실험、Morris수미궁실험검측대서인지행위학개변,면역조직화학법、면역인적법、RT-PCR화매련면역흡부법(ELISA)검측BACE1;ELISA법검측Aβ재당뇨병대서뇌조직중적표체,용도상분석의측정흡광도치.결과 M조대서적전격차수、학습화기억잠복기시간현저연장(P<0.01),주동도피차수현저강저(P<0.01);ELISA법검측Aβ_(1-40)화Aβ_(1-42)현시당뇨병모형서뇌조직내Aβ_(1-40)수평명현증고,종정상조적(64.13±6.76)pg/mg승지모형조적(86.43±7.03)pg/mg(P<0.001);Aβ_(1-42)야명현승고,종정상조적(67.43±5.12 )pg/mg 승지(89.45±5.28) pg/mg (P<0.001);당뇨병모형서뇌조직내BACE1수평현저증고,ELISA법종정상조적(116.46±8.10)pg/mg승지모형조적(158.73±6.24)pg/mg (P<0.001).면역인적법흡광도치종정상조적0.61±0.11승지모형조적1.52±0.16 (P<0.001),RT-PCR법흡광도치종정상조적1.62±0.26승지모형조적3.61±0.32 (P<0.001),면역조직화학법흡광도치종정상조적0.81±0.21승지모형조적2.01±0.36 (P<0.001).BACE1화Aβ표체수평여학습화기억부상관.결론 BACE1화Aβ재당뇨병대서뇌조직표체증고,당뇨병당대사이상증강BACE1화Aβ표체,삼여료아이자해묵병적발병궤제.
Objective To investigate expression of β-site APP-cleaving enzymel(BACE1) and Aβ in brain of diabetes mellitus of Wistar rats,to study pathophysiological mechanism of Alzheimer disease from diabetic metabolic disorder. Methods Animal model of diabetes mellitus was established by streptozocin with intraperitoneal injection. Wistar rats were randomly divided into normal group (N), sham-operation group (S), 4 weeks diabetes mellitus model group (M4), 6 weeks diabetes mellitus model group (M6) and 8 weeks diabetes mellitus model group (M8). Behaviour was tested with Morris water maze and shuttle box test. Expression of Aβ was measured by enzyme linked immunosorbent assay and BACE1 by immunohistochemistry, enzyme linked immunosorbent assay, Western blotting and RT-PCR. The absorbance value was measured by imaging analysis. Results The electric times and latancy of memory and study were more increased in model group than that in N and S group but the times of escape more decreased(P<0.01). The expression of Aβ_(1-40) increased from (64.13±6.76)pg/mg in normal group to (86.43±7.03)pg/mg in model group by ELISA(P<0.001) and Aβ_(1-42) from (67.43±5.12 )pg/mg in normal group to (89.45±5.28) pg/mg (P<0.001) in model group. The expression of BACE1 increased from (116.46±8.10)pg/mg in normal group to (158.73±6.24)pg/mg in model group by ELISA and from 0.61±0.11 in normal group to 1.52±0.16 by Western blotting absorbance valule and from 1.62±0.26 in normal group to 3.61±0.32 by RT-PCR absorbance valule and from 0.81±0.21 in normal group to 2.01±0.36 by immunohistochemistry absorbance valule (P<0.001). The expression of BACE1 and Aβ in MT group was higher than that of in N and S group (P<0.01). The level of BACE1 and Aβ had positive correlation with cognitive impairment.Conclusion The expression of BACE1 and Aβ is increased in diabetes mellitus rats. Diabetes mellitus contributes to Alzheimer diseases that.