滋阴方与温阳方对鱼藤酮诱导PC12细胞损伤的保护作用  

Protection Effects of "Zi-yin" and "Wen-yang" Prescription on PC12 Cell Damage Induced by Rotenone

作　　者：叶承莉[1] 白群华[2] 苏咏梅[2] 唐勇[1] 高洁莹[2] 肖红 贾燕[2] YE Chengli;BAI Qunhua;SHU Yongmei;TANG Yong;GAO Jieying;XIAO Hong;JIA Yan(TCM Orthopedic Hospital of Chongqing City,Chongqing China 400012;Chongqing Medical University,Chongqing China 400016)

机构地区：[1]重庆市中医骨科医院,重庆400012 [2]重庆医科大学,重庆400016

出　　处：《中医学报》2018年第11期2164-2171,共8页Acta Chinese Medicine

基　　金：重庆市渝中区科技计划基金项目(20160121)

摘　　要：目的:探索滋阴方与温阳方(六味地黄丸与肾气丸)对鱼藤酮诱导PC12细胞损伤的保护作用。方法:以鱼藤酮诱导PC12肾上腺髓质嗜铬细胞瘤细胞损伤为模型,观察滋阴方与温阳方(六味地黄丸与肾气丸)对细胞活力、细胞形态、活性氧的产生、凋亡率、ATP的产生以及酪氨酸羟化酶产生的影响作用。结果:滋阴方与温阳方可明显保护鱼藤酮64μg·L-1诱导的PC12细胞的损伤。滋阴方与温阳方可在质量浓度为0. 02 g·m L-1时明显保护鱼藤酮导致的细胞活力的降低,分别使细胞活力由鱼藤酮单独作用时的0. 65±0. 04增加至0. 86±0. 05(P <0. 05)与0. 89±0. 04(P <0. 05);保护鱼藤酮导致的细胞形态的改变,使缩短的突起长度恢复,并减少细胞的收缩变圆程度。滋阴方与温阳方可在质量浓度为0. 01 g·m L-1和0. 02 g·m L-1时,减少鱼藤酮导致的细胞活性氧的产生,使活性氧由鱼藤酮单独作用时的136. 4±9. 3分别减少至联合滋阴方时的(106. 5±7. 3)(P <0. 05)与(63. 6±8. 2)(P <0. 05)和联合温阳方时的98. 2±7. 4(P <0. 05)与58. 2±6. 5(P <0. 05);减少细胞凋亡,使凋亡率由鱼藤酮单独作用时的15. 2%±0. 6%分别减少至联合滋阴方时的11. 5%±0. 5%(P <0. 05)与7. 3%±0. 6%(P <0. 05)和联合温阳方时的10. 3%±0. 4%(P <0. 05)和6. 5%±0. 4%(P <0. 05);增加鱼藤酮导致的细胞内ATP含量的减少(减少至空白对照组的60. 6%±0. 6%),滋阴方使ATP含量增加至空白对照组的85. 4%±5. 7%(P <0. 05)和92. 3%±5. 6%(P <0. 05),而温阳方分别使细胞内ATP含量增加至空白对照组的86. 3%±6. 7%(P <0. 05)和95. 2%±5. 8%(P <0. 05)。另外,滋阴方与温阳方在质量浓度0. 02g·m L-1处理时,保护鱼藤酮导致的细胞内酪氨酸羟化酶(tyrosine hydroxylase,TH)荧光强度减少(减少至空白对照组的66. 6%±7. 6%),分别使细胞内TH荧光强度增加至空白对照组的87. 2%±7. 8%(P <0. 05)和89. 3%±6. 9%(P <0. 05)。结论:滋阴方与温阳方(六味地�Objective: To investigate the protective effects of the zi-yin and wen-yang prescription on PC12 cells damage induced by rotenone. Methods: The injury of PC12 adrenal medullary pheochromocytoma cells induced by rotenone was used as a model to observe the cell viability,cell morphology,reactive oxygen species production and apoptosis rate of zi-yin prescription and wen-yang prescription ( Liuwei Dihuang Pill and Shenqi Pill) , the production of ATP and the influence of the production of tyrosine hydroxylase. Results: zi-yin and wen-yang prescription could significantly protect PC12 cells from rotenone 64 μg·L^-1 . zi-yin prescription and wen-yang prescription can significantly protect the cell viability caused by rotenone at a concentration of 0. 02 g·mL^-1, and increase the cell viability from ( 0. 65 ± 0. 04) when the rotenone acts alone to ( 0. 86 ± 0. 05) ( P < 0. 05) and ( 0. 89 ± 0. 04) ( P < 0. 05) . They can protect the changes in cell morphology caused by rotenone,shorten the protuberance recovery time,and reduce cell shrinkage and roundness. Zi-yin and wen-yang prescription can reduce the production of reactive oxygen species caused by rotenone at concentrations of 0. 01 g·mL^-1 and 0. 02 g·mL^-1, respectively,The reactive oxygen species reduced from 136. 4 ± 9. 3 to 106. 5 ± 7. 3 ( P < 0. 05) and 63. 6 ± 8. 2 ( P < 0. 05) when rotenone alone acts,and to 98. 2 ± 7. 4 ( P < 0. 05) and 58. 2 ± 6. 5 ( P < 0. 05) when combined with zi-yin prescription. The apoptotic rate increased from 15. 2% ± 0. 6% to 11. 5% ± 0. 5% ( P < 0. 05) and 7. 3% ± 0. 6% when rotenone was used alone ( P < 0. 05) and to 10. 3% ± 0. 4% ( P < 0. 05) and 6. 5% ± 0. 4% when combined with zi-yin and wen-yang prescription ( P < 0. 05) . The intracellular ATP content caused by rotenone increased ( reduced to 60. 6% ± 0. 6% of the blank control group) . Zi-yin prescription increased the ATP content to 85. 4% ± 5. 7% ( P < 0. 05) and 92. 3% ± 5. 6% ( P < 0. 05) in the blank control group,while wen-yang prescription incr

关 键 词：滋阴方 温阳方 六味地黄丸 肾气丸 PC12肾上腺髓质嗜铬细胞瘤细胞 鱼藤酮 

分 类 号：R285.5[医药卫生—中药学]