中国药理学与毒理学杂志
中國藥理學與毒理學雜誌
중국약이학여독이학잡지
CHINESE JOURNAL OF PHARMACOLOGY AND TOXICOLOGY
2014年
2期
194-198
,共5页
郭卫伟%常丽俊%丁勇%李欢%葛翠翠%王海洋%张勤丽%牛侨
郭衛偉%常麗俊%丁勇%李歡%葛翠翠%王海洋%張勤麗%牛僑
곽위위%상려준%정용%리환%갈취취%왕해양%장근려%우교
纳米氧化铝%线粒体%自噬
納米氧化鋁%線粒體%自噬
납미양화려%선립체%자서
nano-alu mina%mitochondrion%autophagy
目的:探讨纳米氧化铝对 ICR 小鼠神经细胞线粒体形态及功能的损伤作用。方法健康成年ICR 小鼠分别滴鼻暴露纳米氧化铝(<50 n m)25,50和75 mg·kg -1,连续1个月。电镜观察海马 CA3区神经细胞线粒体的超微结构,并测量线粒体直径;采用定磷法测定脑皮质 Na +-K +-ATPase 与 Ca2+-Mg2+-ATPase 活力;Western 蛋白质印迹法检测细胞色素 c 氧化酶亚单位Ⅳ(COXⅣ),Bcl-2结合蛋白1(Beclin 1)和微管相关蛋白1轻链3Ι蛋白(LC3Ι)和 LC3Ⅱ蛋白的表达。结果与正常对照组相比,50 mg·kg -1组海马 CA3区神经元线粒体明显肿胀,内嵴结构排列稀疏,核周线粒体呈空泡化;线粒体平均直径明显增加〔(0.49±0.02)μm,P<0.05〕;75 mg·kg -1组线粒体大多呈微球形,嵴结构排列紧密,线粒体平均直径显著降低〔(0.36±0.02)μm,P<0.05〕。脑皮质线粒体酶活力随着染毒剂量的增加而降低,与正常对照相比,50和75 mg·kg -1组的 Na +-K +-ATPase 与 Ca2+-Mg2+-ATPase 活力显著降低,分别为6.37±0.22,5.48±1.53和3.21±0.99,(3.28±0.15)kU·g -1蛋白(P <0.05);50和75 mg·kg -1组的 Ca2+-Mg2+-ATPase 活力显著降低(P<0.05)。75 mg·kg -1组 COXⅣ蛋白表达量为1.35±0.66,显著低于其他2个剂量组(P <0.05)。与正常对照相比,75 mg·kg -1组 Beclin 表达量为2.23±0.20,自噬特异蛋白LC3Ⅱ/LC3Ⅰ的比例为0.45±0.10,均显著高于正常对照组(P<0.05)。结论线粒体功能紊乱可能是纳米氧化铝神经毒性机制之一,而神经细胞可能通过自噬清除受损线粒体。
目的:探討納米氧化鋁對 ICR 小鼠神經細胞線粒體形態及功能的損傷作用。方法健康成年ICR 小鼠分彆滴鼻暴露納米氧化鋁(<50 n m)25,50和75 mg·kg -1,連續1箇月。電鏡觀察海馬 CA3區神經細胞線粒體的超微結構,併測量線粒體直徑;採用定燐法測定腦皮質 Na +-K +-ATPase 與 Ca2+-Mg2+-ATPase 活力;Western 蛋白質印跡法檢測細胞色素 c 氧化酶亞單位Ⅳ(COXⅣ),Bcl-2結閤蛋白1(Beclin 1)和微管相關蛋白1輕鏈3Ι蛋白(LC3Ι)和 LC3Ⅱ蛋白的錶達。結果與正常對照組相比,50 mg·kg -1組海馬 CA3區神經元線粒體明顯腫脹,內嵴結構排列稀疏,覈週線粒體呈空泡化;線粒體平均直徑明顯增加〔(0.49±0.02)μm,P<0.05〕;75 mg·kg -1組線粒體大多呈微毬形,嵴結構排列緊密,線粒體平均直徑顯著降低〔(0.36±0.02)μm,P<0.05〕。腦皮質線粒體酶活力隨著染毒劑量的增加而降低,與正常對照相比,50和75 mg·kg -1組的 Na +-K +-ATPase 與 Ca2+-Mg2+-ATPase 活力顯著降低,分彆為6.37±0.22,5.48±1.53和3.21±0.99,(3.28±0.15)kU·g -1蛋白(P <0.05);50和75 mg·kg -1組的 Ca2+-Mg2+-ATPase 活力顯著降低(P<0.05)。75 mg·kg -1組 COXⅣ蛋白錶達量為1.35±0.66,顯著低于其他2箇劑量組(P <0.05)。與正常對照相比,75 mg·kg -1組 Beclin 錶達量為2.23±0.20,自噬特異蛋白LC3Ⅱ/LC3Ⅰ的比例為0.45±0.10,均顯著高于正常對照組(P<0.05)。結論線粒體功能紊亂可能是納米氧化鋁神經毒性機製之一,而神經細胞可能通過自噬清除受損線粒體。
목적:탐토납미양화려대 ICR 소서신경세포선립체형태급공능적손상작용。방법건강성년ICR 소서분별적비폭로납미양화려(<50 n m)25,50화75 mg·kg -1,련속1개월。전경관찰해마 CA3구신경세포선립체적초미결구,병측량선립체직경;채용정린법측정뇌피질 Na +-K +-ATPase 여 Ca2+-Mg2+-ATPase 활력;Western 단백질인적법검측세포색소 c 양화매아단위Ⅳ(COXⅣ),Bcl-2결합단백1(Beclin 1)화미관상관단백1경련3Ι단백(LC3Ι)화 LC3Ⅱ단백적표체。결과여정상대조조상비,50 mg·kg -1조해마 CA3구신경원선립체명현종창,내척결구배렬희소,핵주선립체정공포화;선립체평균직경명현증가〔(0.49±0.02)μm,P<0.05〕;75 mg·kg -1조선립체대다정미구형,척결구배렬긴밀,선립체평균직경현저강저〔(0.36±0.02)μm,P<0.05〕。뇌피질선립체매활력수착염독제량적증가이강저,여정상대조상비,50화75 mg·kg -1조적 Na +-K +-ATPase 여 Ca2+-Mg2+-ATPase 활력현저강저,분별위6.37±0.22,5.48±1.53화3.21±0.99,(3.28±0.15)kU·g -1단백(P <0.05);50화75 mg·kg -1조적 Ca2+-Mg2+-ATPase 활력현저강저(P<0.05)。75 mg·kg -1조 COXⅣ단백표체량위1.35±0.66,현저저우기타2개제량조(P <0.05)。여정상대조상비,75 mg·kg -1조 Beclin 표체량위2.23±0.20,자서특이단백LC3Ⅱ/LC3Ⅰ적비례위0.45±0.10,균현저고우정상대조조(P<0.05)。결론선립체공능문란가능시납미양화려신경독성궤제지일,이신경세포가능통과자서청제수손선립체。
OBJECTIVE To explore the potential neurotoxicity of nano-alu mina (<50 n m)in vivo, we treated the ICR mouse with the nano-alu mina to investigate the mitochondrial da mage of nerve cells on morphology and function.METHODS Adult male mice were exposed to nano-alu mina (<50 n m)of 0,25,50 and 75 mg·kg -1 by nasal instillation for 1 month.Then we observed the mitochondrial ultra-structure of the nerve cells in CA3 region of hippoca mpus,and measured the mean dia meter in every group.The activities of Na +-K +-ATPase and Ca2 +-Mg2 +-ATPase were tested by the determination of the inorganic phosphorus,which was the deco mposition product of ATPase.Western blot analysis was used to detect the expression of COX-Ⅳ,Beclin1 ,LC3Ιand LC3Ⅱ.RESULTS Co mpared with 0 and 25 mg·kg -1 groups exposed to Al2 O3 nanopartilces (Al2 O3 NPs),the mitochondria of CA3 region in hip-poca mpus in 50 mg·kg -1 group beca me ede matous and swollen with sparse and broken cristae sur-rounding the nuclear,and the mean dia meter was higher(0.49 ±0.02 μm,P <0.05).But co mpared with 50 mg·kg -1 group,the mitochondria in 75 mg·kg -1 group beca me s maller with inner cristae of high density,and the mean dia meter was lower(0.36 ±0.02 μm,P<0.05).The enzy me activity of the mito-chondria in cerebral cortex decreased dose-dependently with exposure,the activities of Na +-K +-ATPase in 50 and 75 mg·kg -1 groups(6.37 ±0.22 kU·g -1 protein,5.48 ±1 .53 kU·g -1 protein)and Ca2 +-Mg2 +-ATPase in 50 and 75 mg·kg -1 groups (3.21 ±0.99 kU·g -1 protein,3.28 ±0.15 kU·g -1 protein)were lower than the 0 mg·kg -1 group(P<0.05).Meanwhile,the Ca2 +-Mg2 +-ATPase in 50 and 75 mg·kg -1 groups showed lower activities in co mparison with the 25 mg·kg -1 group.The 75 mg·kg -1 group expressed higher level of the COX-Ⅳ protein 1 .35 ±0.66(P<0.05)than other groups.Both expression of Beclin1 protein and rate of LC3Ⅱ/LC3Ⅰin 75 mg·kg -1 group were more than the 0 mg·kg -1 group. CONCLUSION The mitochondrial dysfunction may be the potential neurotoxicity of nano-alu mina,and the da maged mitochondria were cleared by autophagy.