20种中草药及开心散对PC12和SH-SY5Y细胞的保护作用
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摘要
目的
选择20种中草药、开心散复方、参远苷复合物及其活性成分观察其对PC12细胞和SH-SY5Y细胞增殖的影响,并探讨其对H2O2、谷氨酸(glutamate, GLU)、皮质醇诱导PC12细胞和SH-SY5Y细胞损伤的保护作用,为寻找神经细胞保护作用新药提供实验依据。
方法
体外培养PC12细胞和SH-SY5Y细胞,应用不同浓度的H2O2、GLU、皮质醇分别建立神经细胞损伤模型,采用MTT法检测细胞存活率。结果
罗勒、虎爪豆、诃子、天麻、参远苷复合物等具有明显促进PC12细胞增殖作用(P<0.05)；罗勒、灯油藤、甘草、虎爪豆、诃子、天麻、远志总苷及人参总皂苷具有明显的促进SH-SY5Y细胞增殖作用(P<0.05)；在H2O2100μmol/L损伤模型中,药物组和模型组相比,罗勒、仙人掌、芦笋、灯油藤、诃子、益智仁、天麻、参远苷复合物、远志总苷等能显著减轻H2O2诱导的PC12细胞损伤作用(P<0.05)；积雪草、指甲花、罗勒、芦笋、甘草、虎爪豆、诃子、葡萄、益智仁天麻、参远苷复合物、远志总苷等能显著减轻H202诱导的SH-SY5Y细胞损伤作用(P<0.05)；在GLU 30mmol/L诱导PC12细胞损伤,与模型组比较,凤仙花、仙人掌、虎爪豆、诃子、益智仁、天麻、参远苷复合物及人参总皂苷等均显示出损伤保护作用(P<0.05)；在GLU 30mmol/L诱导SH-SY5Y细胞损伤,与模型组比较,积雪草、指甲花、罗勒、芦笋、灯油藤、甘草、虎爪豆、肉豆蔻、诃子、益智仁、天麻、参远苷复合物及远志总苷表现出显著的损伤保护活性(P<0.05)；在皮质醇200μmol/L诱导PC12细胞损伤模型中,药物组与模型组相比,积雪草、指甲花、仙人掌、芦笋、灯油藤、虎爪豆、葡萄、益智仁、天麻、参远苷复合物、远志总苷和人参总皂苷能显著的减轻皮质醇诱导的PC12细胞损伤作用(P<0.05)；在皮质醇100μmol/L诱导SH-SY5Y细胞损伤模型中,药物组与模型组相比,积雪草、指甲花、罗勒、芦笋、灯油藤、甘草、诃子、益智仁天麻、参远苷复合物和远志总苷能显著的降低皮质醇对SH-SY5Y细胞的损伤作用。
结论
对PC12细胞和SH-SY5Y细胞均具有增殖作用的药物为：罗勒、虎爪豆、诃子、天麻、参远苷复合物和远志总苷；具有同时对抗H2O2诱导PC12细胞和SH-SY5Y细胞损伤作用的药物为：积雪草、指甲花、罗勒、芦笋、诃子、益智仁、天麻、参远苷复合物、远志总苷及人参总皂苷；同时具有减弱GLU诱导PC12细胞和SH-SY5Y细胞损伤作用的药物为：指甲花、芦笋、甘草、虎爪豆、诃子、益智仁、天麻及参远苷复合物；具有对抗皮质醇诱导PC12细胞和SH-SY5Y细胞损伤作用的药物为：积雪草、指甲花、芦笋、灯油藤、益智仁、天麻、参远苷复合物和远志总苷。其中,益智仁、天麻、参远苷复合物在四个模型中均表现出较强的细胞保护作用。本研究结果对神经退行性疾病和抑郁症药物的开发和选择提供了初步的实验依据。
Objective
To investigate the protective effect of 46 extracts from 20 plants and Kai-Xin-San in PC 12 cells and SH-SY5Y cells injured by H2O2, glutamate (GLU) and hydrocortisone.
Method
Different concentrations of H2O2, GLU and hydrocortisone were used to establish the toxicity models on PC 12 cells and SH-SY5Y cells. Cell viability was determined by MTT assay.
Results
The extracts of Ocimum basilicum, Mucuna pruriens, Terminalia chebula, Alpiniaoxyghyl-lamiq and a mixtured substance from Panax ginseng and Radix Polygalae had significantly proliferative effects on PC 12 cells (P<0.05); The extracts of Ocimum basilicum, Celastrus paniculata, Glycyrrhiza glabra, Mucuna pruriens, Terminalia chebula, Panax ginseng and Radix Polygalae facilitate the proliferation of SH-SY5Y cells significantly (P<0.05). Additionally, the extracts of Ocimum basilicum, Opuntia delineii, Asparagus officinalis, Celastrus paniculata, Terminalia chebula, Gastrodia elata, Alpiniaoxyghyl-lamiq, a mixtured substance from Panax ginseng and Radix Polygalae, Radix Polygalae showed markedly protective effects against H2O2-induced PC 12 cells death; The extracts of Centilla asiatica, Lawsonia alba, Ocimum basilicum, Asparagus officinalis, Glycyrrhiza glabra, Mucuna pruriens, Terminalia chebula, Vitis vinifera, Gastrodia elata, Alpiniaoxyghyl-lamiq, a mixtured substance from Panax ginseng and Radix Polygalae, Radix Polygalae showed markedly protective effects against H2O2-induced SH-SY5Y cells death. Simiarly results were observed in GLU injury model, which showed that the extracts of Lawsonia alba, Opuntia delineii, Mucuna pruriens, Myrisica fragrans, Terminalia chebula, a mixtured substance from Panax ginseng and Radix Polygalae, Radix Polygalae attenuated GLU-induced toxicity obviously on PC 12 cells. And on SH-SY5Y cells by GLU-induced toxiously, the extracts of Centilla asiatica, Lawsonia alba, Ocimum basilicum, Asparagus officinalis, Celastrus paniculata, Glycyrrhiza glabra, Mucuna pruriens, Myrisica fragrans, Terminalia chebula, Gastrodia elata, Alpiniaoxyghyl-lamiq, a mixtured substance from Panax ginseng and Radix Polygalae, Radix Polygalae attenuated GLU-induced toxicity obviously. And on the hydrocortisone injury model, the extracts of Centilla asiatica, Lawsonia alba, Opuntia delineii, Asparagus officinalis, Celastrus paniculata, Mucuna pruriens, Vitis vinifera, Gastrodia elata, Alpiniaoxyghyl-lamiq, a mixtured substance from Panax ginseng and Radix Polygalae, Panax ginseng and Radix Polygalae showed markedly protective effects against hydrocortisone-induced PC12 cells death; Simiarly results were observed to injuryed SH-SY5Y cells by hydrocortisone, the extracts of Centilla asiatica, Lawsonia alba, Ocimum basilicum, Asparagus officinalis, Asparagus officinalis, Mucuna pruriens, Terminalia chebula, Gastrodia elata, Alpiniaoxyghyl-lamiq, a mixtured substance from Panax ginseng and Radix Polygalae, Panax ginseng showed markedly protective effects against hydrocortisone- induced SH-SY5Y cells death.
Conclusion
The extracts of Ocimum basilicum, Mucuna pruriens, Terminalia chebula, Gastrodia elata, a mixtured substance from Panax ginseng and Radix Polygalae, Radix Polygalae have the promoting effect on the proliferation of PC 12 cells and SH-SY5Y cells. Centilla asiatica, Lawsonia alba, Ocimum basilicum, Asparagus officinalis, Terminalia chebula, Gastrodia elata, Alpiniaoxyphyl-lamiq, a mixtured substance from Panax ginseng and Radix Polygalae, Panax ginseng and Radix Polygalae extracts possess significant antioxidative activities. And the extracts from Lawsonia alba, Asparagus officinalis, Glycyrrhiza glabra, Mucuna pruriens, Terminalia chebula, Gastrodia elata, Alpiniaoxyphyl-lamiq, a mixtured substance from Panax ginseng and Radix Polygalae can decreasing the damage induced by GLU in PC12 cells and SH-SY5Y cells. The extracts of Centilla asiatica, Lawsonia alba, Asparagus officinalis, Celastrus prostrate, Gastrodia elata, Alpiniaoxyphyl-lamiq, a mixtured substance from Panax ginseng and Radix Polygalae, Radix Polygalae can decreasing the damage induced by hydrocortisone in PC 12 cells and SH-SY5Y cells. Among all the extracts, Gastrodia elata, Alpiniaoxyphyl-lamiq, a mixtured substance from Panax ginseng and Radix Polygalae had protective effect on all H2O2, GLU and hydrocortisone induced neurotoxicity, with proliferative effect on PC-12 cells and SH-SY5Y cells. The results of this s tudy provide a preliminary experimental basis of screening and developing new agents for ND and depression.
选择20种中草药、开心散复方、参远苷复合物及其活性成分观察其对PC12细胞和SH-SY5Y细胞增殖的影响,并探讨其对H2O2、谷氨酸(glutamate, GLU)、皮质醇诱导PC12细胞和SH-SY5Y细胞损伤的保护作用,为寻找神经细胞保护作用新药提供实验依据。
方法
体外培养PC12细胞和SH-SY5Y细胞,应用不同浓度的H2O2、GLU、皮质醇分别建立神经细胞损伤模型,采用MTT法检测细胞存活率。结果
罗勒、虎爪豆、诃子、天麻、参远苷复合物等具有明显促进PC12细胞增殖作用(P<0.05)；罗勒、灯油藤、甘草、虎爪豆、诃子、天麻、远志总苷及人参总皂苷具有明显的促进SH-SY5Y细胞增殖作用(P<0.05)；在H2O2100μmol/L损伤模型中,药物组和模型组相比,罗勒、仙人掌、芦笋、灯油藤、诃子、益智仁、天麻、参远苷复合物、远志总苷等能显著减轻H2O2诱导的PC12细胞损伤作用(P<0.05)；积雪草、指甲花、罗勒、芦笋、甘草、虎爪豆、诃子、葡萄、益智仁天麻、参远苷复合物、远志总苷等能显著减轻H202诱导的SH-SY5Y细胞损伤作用(P<0.05)；在GLU 30mmol/L诱导PC12细胞损伤,与模型组比较,凤仙花、仙人掌、虎爪豆、诃子、益智仁、天麻、参远苷复合物及人参总皂苷等均显示出损伤保护作用(P<0.05)；在GLU 30mmol/L诱导SH-SY5Y细胞损伤,与模型组比较,积雪草、指甲花、罗勒、芦笋、灯油藤、甘草、虎爪豆、肉豆蔻、诃子、益智仁、天麻、参远苷复合物及远志总苷表现出显著的损伤保护活性(P<0.05)；在皮质醇200μmol/L诱导PC12细胞损伤模型中,药物组与模型组相比,积雪草、指甲花、仙人掌、芦笋、灯油藤、虎爪豆、葡萄、益智仁、天麻、参远苷复合物、远志总苷和人参总皂苷能显著的减轻皮质醇诱导的PC12细胞损伤作用(P<0.05)；在皮质醇100μmol/L诱导SH-SY5Y细胞损伤模型中,药物组与模型组相比,积雪草、指甲花、罗勒、芦笋、灯油藤、甘草、诃子、益智仁天麻、参远苷复合物和远志总苷能显著的降低皮质醇对SH-SY5Y细胞的损伤作用。
结论
对PC12细胞和SH-SY5Y细胞均具有增殖作用的药物为：罗勒、虎爪豆、诃子、天麻、参远苷复合物和远志总苷；具有同时对抗H2O2诱导PC12细胞和SH-SY5Y细胞损伤作用的药物为：积雪草、指甲花、罗勒、芦笋、诃子、益智仁、天麻、参远苷复合物、远志总苷及人参总皂苷；同时具有减弱GLU诱导PC12细胞和SH-SY5Y细胞损伤作用的药物为：指甲花、芦笋、甘草、虎爪豆、诃子、益智仁、天麻及参远苷复合物；具有对抗皮质醇诱导PC12细胞和SH-SY5Y细胞损伤作用的药物为：积雪草、指甲花、芦笋、灯油藤、益智仁、天麻、参远苷复合物和远志总苷。其中,益智仁、天麻、参远苷复合物在四个模型中均表现出较强的细胞保护作用。本研究结果对神经退行性疾病和抑郁症药物的开发和选择提供了初步的实验依据。
Objective
To investigate the protective effect of 46 extracts from 20 plants and Kai-Xin-San in PC 12 cells and SH-SY5Y cells injured by H2O2, glutamate (GLU) and hydrocortisone.
Method
Different concentrations of H2O2, GLU and hydrocortisone were used to establish the toxicity models on PC 12 cells and SH-SY5Y cells. Cell viability was determined by MTT assay.
Results
The extracts of Ocimum basilicum, Mucuna pruriens, Terminalia chebula, Alpiniaoxyghyl-lamiq and a mixtured substance from Panax ginseng and Radix Polygalae had significantly proliferative effects on PC 12 cells (P<0.05); The extracts of Ocimum basilicum, Celastrus paniculata, Glycyrrhiza glabra, Mucuna pruriens, Terminalia chebula, Panax ginseng and Radix Polygalae facilitate the proliferation of SH-SY5Y cells significantly (P<0.05). Additionally, the extracts of Ocimum basilicum, Opuntia delineii, Asparagus officinalis, Celastrus paniculata, Terminalia chebula, Gastrodia elata, Alpiniaoxyghyl-lamiq, a mixtured substance from Panax ginseng and Radix Polygalae, Radix Polygalae showed markedly protective effects against H2O2-induced PC 12 cells death; The extracts of Centilla asiatica, Lawsonia alba, Ocimum basilicum, Asparagus officinalis, Glycyrrhiza glabra, Mucuna pruriens, Terminalia chebula, Vitis vinifera, Gastrodia elata, Alpiniaoxyghyl-lamiq, a mixtured substance from Panax ginseng and Radix Polygalae, Radix Polygalae showed markedly protective effects against H2O2-induced SH-SY5Y cells death. Simiarly results were observed in GLU injury model, which showed that the extracts of Lawsonia alba, Opuntia delineii, Mucuna pruriens, Myrisica fragrans, Terminalia chebula, a mixtured substance from Panax ginseng and Radix Polygalae, Radix Polygalae attenuated GLU-induced toxicity obviously on PC 12 cells. And on SH-SY5Y cells by GLU-induced toxiously, the extracts of Centilla asiatica, Lawsonia alba, Ocimum basilicum, Asparagus officinalis, Celastrus paniculata, Glycyrrhiza glabra, Mucuna pruriens, Myrisica fragrans, Terminalia chebula, Gastrodia elata, Alpiniaoxyghyl-lamiq, a mixtured substance from Panax ginseng and Radix Polygalae, Radix Polygalae attenuated GLU-induced toxicity obviously. And on the hydrocortisone injury model, the extracts of Centilla asiatica, Lawsonia alba, Opuntia delineii, Asparagus officinalis, Celastrus paniculata, Mucuna pruriens, Vitis vinifera, Gastrodia elata, Alpiniaoxyghyl-lamiq, a mixtured substance from Panax ginseng and Radix Polygalae, Panax ginseng and Radix Polygalae showed markedly protective effects against hydrocortisone-induced PC12 cells death; Simiarly results were observed to injuryed SH-SY5Y cells by hydrocortisone, the extracts of Centilla asiatica, Lawsonia alba, Ocimum basilicum, Asparagus officinalis, Asparagus officinalis, Mucuna pruriens, Terminalia chebula, Gastrodia elata, Alpiniaoxyghyl-lamiq, a mixtured substance from Panax ginseng and Radix Polygalae, Panax ginseng showed markedly protective effects against hydrocortisone- induced SH-SY5Y cells death.
Conclusion
The extracts of Ocimum basilicum, Mucuna pruriens, Terminalia chebula, Gastrodia elata, a mixtured substance from Panax ginseng and Radix Polygalae, Radix Polygalae have the promoting effect on the proliferation of PC 12 cells and SH-SY5Y cells. Centilla asiatica, Lawsonia alba, Ocimum basilicum, Asparagus officinalis, Terminalia chebula, Gastrodia elata, Alpiniaoxyphyl-lamiq, a mixtured substance from Panax ginseng and Radix Polygalae, Panax ginseng and Radix Polygalae extracts possess significant antioxidative activities. And the extracts from Lawsonia alba, Asparagus officinalis, Glycyrrhiza glabra, Mucuna pruriens, Terminalia chebula, Gastrodia elata, Alpiniaoxyphyl-lamiq, a mixtured substance from Panax ginseng and Radix Polygalae can decreasing the damage induced by GLU in PC12 cells and SH-SY5Y cells. The extracts of Centilla asiatica, Lawsonia alba, Asparagus officinalis, Celastrus prostrate, Gastrodia elata, Alpiniaoxyphyl-lamiq, a mixtured substance from Panax ginseng and Radix Polygalae, Radix Polygalae can decreasing the damage induced by hydrocortisone in PC 12 cells and SH-SY5Y cells. Among all the extracts, Gastrodia elata, Alpiniaoxyphyl-lamiq, a mixtured substance from Panax ginseng and Radix Polygalae had protective effect on all H2O2, GLU and hydrocortisone induced neurotoxicity, with proliferative effect on PC-12 cells and SH-SY5Y cells. The results of this s tudy provide a preliminary experimental basis of screening and developing new agents for ND and depression.
引文
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