3-羟基丁酸衍生物对PC12细胞凋亡的保护作用
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摘要
目的观察3-羟基丁酸甲酯和3-羟基丁酸乙酯对PC12缺血清诱导凋亡细胞的保护作用。方法化学法降解聚羟基丁酸酯制备3-羟基丁酸甲酯和3-羟基丁酸乙酯。培养PC12细胞,以含血清培养组细胞为阴性对照组,缺血清培养组细胞为阳性对照组,以在缺血清条件下添加不同浓度3-羟基丁酸甲酯或3-羟基丁酸乙酯细胞为样品组,通过形态学观察法、噻唑蓝(MTT)法检测各组细胞形态及活性变化。结果与阴性对照组比较,阳性对照组细胞出现大量凋亡,形态变小变细,活性降低。样品处理组细胞活性不同程度提高,低浓度(0.01和0.001 mg·m L-1)3-羟基丁酸甲酯和3-羟基丁酸乙酯对凋亡细胞有保护作用,浓度为1 mg·m L-1时效果最好。结论通过化学法能够制备高纯度3-羟基丁酸甲酯和3-羟基丁酸乙酯,上述两种样品对缺血清诱导凋亡的PC12细胞有保护作用。
Objective To investigate the protective effect of 3-hydroxybutyrate derivate( methyl-3-hydroxybutyrate and ethyl-3-hydroxybutyrate) on the apoptosis of PC12 induced by FBS deficiency. Methods Methyl-3-hydroxybutyrate and ethyl-3-hydroxybutyrate were prepared by chemical degradation. PC12 cells were divided into different groups: medium with FBS as negative control,medium without FBS as positive control,medium with different concentrations of methyl-3-hydroxybutyrate or ethyl-3-hydroxybutyrate without FBS as sample groups. The shape and viability of cells in each group were analyzed by optical microscope and MTT. Results Compared with the negative control,cells in the positive control group demonstrated a large number of apoptosis,smaller and thinner morphology,and lower activity. However,the activity of cells was improved in the sample groups. Low concentration( 0. 01 and 0. 001 mg · m L- 1) of methyl-3-hydroxybutyrate and ethyl-3-hydroxybutyrate showed a protective effect on cell apoptosis,and 1 mg · m L- 1had the best protection effect. Conclusion High purity methyl-3-hydroxybutyrate and ethyl-3-hydroxybutyrate can be produced by chemical method,and both chemicals present good effect on protecting the PC12 from apoptosis.
Objective To investigate the protective effect of 3-hydroxybutyrate derivate( methyl-3-hydroxybutyrate and ethyl-3-hydroxybutyrate) on the apoptosis of PC12 induced by FBS deficiency. Methods Methyl-3-hydroxybutyrate and ethyl-3-hydroxybutyrate were prepared by chemical degradation. PC12 cells were divided into different groups: medium with FBS as negative control,medium without FBS as positive control,medium with different concentrations of methyl-3-hydroxybutyrate or ethyl-3-hydroxybutyrate without FBS as sample groups. The shape and viability of cells in each group were analyzed by optical microscope and MTT. Results Compared with the negative control,cells in the positive control group demonstrated a large number of apoptosis,smaller and thinner morphology,and lower activity. However,the activity of cells was improved in the sample groups. Low concentration( 0. 01 and 0. 001 mg · m L- 1) of methyl-3-hydroxybutyrate and ethyl-3-hydroxybutyrate showed a protective effect on cell apoptosis,and 1 mg · m L- 1had the best protection effect. Conclusion High purity methyl-3-hydroxybutyrate and ethyl-3-hydroxybutyrate can be produced by chemical method,and both chemicals present good effect on protecting the PC12 from apoptosis.
引文
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[2]PLECKO B,STOCKLER I S,SCHOBER E,et al.Oral beta-hydroxybutyrate supplementation in two Patients with hyperinsulinemic hypoglycemia:monitoring of betahydroxybutyrate levels in blood and cerebrospinal fluid,and in the brain by in vivo magnetic resonance spectroscopy[J].Pediatry Res,2002,52(2):301-306.
[3]XIAO X Q,ZHAO Y,CHEN G Q.The effect of 3-hydroxybutyrate and its derivatives on the growth of glial cells[J].Biomaterials,2007,28(25):3608-3616.
[4]ZOU X H,LI H M,WANG S,et al.The effect of 3-hydroxybutyrate methyl ester on learning and memory in mice[J].Biomaterials,2009,30(8):1532-1541.
[5]WU Q,SUN S Q,YU P.Environmental dependence of microbial synthesis of Polyhydroxyalkanoates[J].Acta Polymerica Sinica,2000,1(6):751-756.
[6]DAIKHIN Y,YUDKOFF M.Ketone bodies and brain glutamate and GABA metabolism[J].Dev Neuroseienee-Basel,1998,20(4-5):358-364.
[7]CHENG S,CHEN G Q,LESKI M,et al.The effect of D,L-β-Hydroxybutyric acid on cell death and proliferation in L929 cells[J].Biomaterials,2006,27(20):3758-3765.
[8]SARANSAARI P,OJA S S.Taurine and neural cell damage[J].Amino Acids,2000,19(3-4):509-526.
[9]GREENE L A,TISCHLERA S.Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor[J].Proc Natl Acad Sci USA,1976,73(7):2424-2428.
[10]KERR J F,WYLLIE A H,CURRIE A R.Apoptosis:a basic biological phenomenon with wide ranging implications in tissue kinetics[J].Br J Cancer,1972,26(4):239-257.
[11]MOGI M,OZEKI N,NAKAMURA H T.Dual roles for NFkappa B activation in osteoblastic cells by serum deprivation:osteoblastic apoptosis and cell-cycle arrest[J].Bone,2004,35(2):507-516.