中华神经医学杂志
中華神經醫學雜誌
중화신경의학잡지
CHINESE JOURNAL OF NEUROMEDICINE
2010年
2期
119-123
,共5页
姜黄素%热休克蛋白70%α-突触核蛋白%蛋白酶体活性
薑黃素%熱休剋蛋白70%α-突觸覈蛋白%蛋白酶體活性
강황소%열휴극단백70%α-돌촉핵단백%단백매체활성
Curcumin%Heat shock protein 70%α-synuclein%Activity ofproteasome
目的 探讨姜黄素通过诱导热休克蛋白70(Hsp70)高表达,抑制α-突触核蛋白的异常表达和聚集,促进蛋白酶体系统降解异常α-突触核蛋白对多巴胺能细胞的保护作用.方法 选用大鼠嗜铬细胞瘤细胞株(PCI2细胞),鱼藤酮诱导其损伤建立帕金森病细胞模型,利用姜黄素进行干预:采用MTT法检测细胞活力,荧光酶标仪检测蛋白酶体水解酶活性,Western blot检测Hsp70和α-突触核蛋白的表达,免疫荧光法检测细胞内Hsp70的表达和α-突触核蛋白的聚集.结果 鱼藤酮组PC12细胞活力及蛋白酶体水解酶活性明显降低,Hsp70的表达轻度增加,α-突触核蛋白表达和聚集明显增加,与对照组比较差异有统计学意义(P<0.05).经不同浓度姜黄素预处理4h后与0.1 μmol/L鱼藤酮共同孵育PC12细胞24 h,与鱼藤酮组比较,0.5 μmol/L和1.0 μmol/L姜黄素使细胞活力以及蛋白酶体水解酶活性明显升高,Hsp70表达明显升高,α-突触核蛋白的表达和聚集明显减少,差异有统计学意义(P<0.05);5.0 μmol/L和10 μmol/L姜黄素对鱼藤酮的拮抗作用明显减弱,细胞活力与鱼藤酮组比较差异均无统计学意义(P>0.05),胰蛋白酶、多肽-谷氨酰-多肽水解酶活性与鱼藤酮组比较差异均无统计学意义(P>0.05);10 μmol/L姜黄素组糜蛋白酶样水解酶活性进一步降低,与鱼藤酮组比较差异有统计学意义(P<0.05).结论 低浓度姜黄素能够通过诱导PC12细胞表达Hsp70,诱导蛋白酶体水解酶活性表达,进而抑制α-突触核蛋白的表达和聚集,从而拮抗鱼藤酮诱导的PC12细胞的损伤.
目的 探討薑黃素通過誘導熱休剋蛋白70(Hsp70)高錶達,抑製α-突觸覈蛋白的異常錶達和聚集,促進蛋白酶體繫統降解異常α-突觸覈蛋白對多巴胺能細胞的保護作用.方法 選用大鼠嗜鉻細胞瘤細胞株(PCI2細胞),魚籐酮誘導其損傷建立帕金森病細胞模型,利用薑黃素進行榦預:採用MTT法檢測細胞活力,熒光酶標儀檢測蛋白酶體水解酶活性,Western blot檢測Hsp70和α-突觸覈蛋白的錶達,免疫熒光法檢測細胞內Hsp70的錶達和α-突觸覈蛋白的聚集.結果 魚籐酮組PC12細胞活力及蛋白酶體水解酶活性明顯降低,Hsp70的錶達輕度增加,α-突觸覈蛋白錶達和聚集明顯增加,與對照組比較差異有統計學意義(P<0.05).經不同濃度薑黃素預處理4h後與0.1 μmol/L魚籐酮共同孵育PC12細胞24 h,與魚籐酮組比較,0.5 μmol/L和1.0 μmol/L薑黃素使細胞活力以及蛋白酶體水解酶活性明顯升高,Hsp70錶達明顯升高,α-突觸覈蛋白的錶達和聚集明顯減少,差異有統計學意義(P<0.05);5.0 μmol/L和10 μmol/L薑黃素對魚籐酮的拮抗作用明顯減弱,細胞活力與魚籐酮組比較差異均無統計學意義(P>0.05),胰蛋白酶、多肽-穀氨酰-多肽水解酶活性與魚籐酮組比較差異均無統計學意義(P>0.05);10 μmol/L薑黃素組糜蛋白酶樣水解酶活性進一步降低,與魚籐酮組比較差異有統計學意義(P<0.05).結論 低濃度薑黃素能夠通過誘導PC12細胞錶達Hsp70,誘導蛋白酶體水解酶活性錶達,進而抑製α-突觸覈蛋白的錶達和聚集,從而拮抗魚籐酮誘導的PC12細胞的損傷.
목적 탐토강황소통과유도열휴극단백70(Hsp70)고표체,억제α-돌촉핵단백적이상표체화취집,촉진단백매체계통강해이상α-돌촉핵단백대다파알능세포적보호작용.방법 선용대서기락세포류세포주(PCI2세포),어등동유도기손상건립파금삼병세포모형,이용강황소진행간예:채용MTT법검측세포활력,형광매표의검측단백매체수해매활성,Western blot검측Hsp70화α-돌촉핵단백적표체,면역형광법검측세포내Hsp70적표체화α-돌촉핵단백적취집.결과 어등동조PC12세포활력급단백매체수해매활성명현강저,Hsp70적표체경도증가,α-돌촉핵단백표체화취집명현증가,여대조조비교차이유통계학의의(P<0.05).경불동농도강황소예처리4h후여0.1 μmol/L어등동공동부육PC12세포24 h,여어등동조비교,0.5 μmol/L화1.0 μmol/L강황소사세포활력이급단백매체수해매활성명현승고,Hsp70표체명현승고,α-돌촉핵단백적표체화취집명현감소,차이유통계학의의(P<0.05);5.0 μmol/L화10 μmol/L강황소대어등동적길항작용명현감약,세포활력여어등동조비교차이균무통계학의의(P>0.05),이단백매、다태-곡안선-다태수해매활성여어등동조비교차이균무통계학의의(P>0.05);10 μmol/L강황소조미단백매양수해매활성진일보강저,여어등동조비교차이유통계학의의(P<0.05).결론 저농도강황소능구통과유도PC12세포표체Hsp70,유도단백매체수해매활성표체,진이억제α-돌촉핵단백적표체화취집,종이길항어등동유도적PC12세포적손상.
Objective To explore the protective effects of curcumin on dopaminergic cells by inducing the expression of heat shock protein 70 (Hsp70), inhibiting anormal expression and aggregation of α-synuclein and promoting proteasome system to degrade abnormal α-synuclein. Methods The cellular model of Parkinson's disease was induced by rotenone in the rat pheochromocytoma strain (PC12 cells) and curcumin was also employed to intervene the cell expression. Cell viability was assessed by MTT assay and the activity of the 3 hydrolases in proteasome was measured by the method of fluorescent anzyme immunoassay. Western blotting was employed to detect the expressions of Hsp70 and α-synuclein and immunofluorescence was used to observe the expression of Hsp70 and the aggregation of α-synuclein. Results The cell viability and the activity of proteasome in the rotenone group were decreased dramatically; the expression of Hsp70 was slightly increased while the expression and aggregation of α-synuclein was increased significantly as compared with those in other groups (P<0.05). Pretreatmant with 0.5 μmol/L and 1.0 μmol/L curcumin resulted in significantly increased cell viability and activity ofproteasome in PC12 cells after co-incubation with 0.1 μmol/L rotenone for 24 h compared with the rotenone group; the expression of Hsp70 was increased significantly and significantly decreased expression and aggregation of α-synuclein was found (P<0.05). The protective effects of 5.0 μmol/L and 10 μmol/L curcumin against the rotenone-induced injury in PC12 cells decreased apparently; when the curcumin reached 10 μmol/L, the activity of chymotrypsin in proteasome statistically lower than that in the rotenone group; the activity of the other 2 enzymes in proteasome were not significantly different from that in the rotenone group (P>0.05). Conclusion Low concentration of curcumin can induce higher expression of Hsp70 and higher activity of proteasome to inhibit the anormal expression and aggregation of α-synuclein in PC 12 cells, thus it can alleviate the rotenone-induced injury in PC 12 cells.
