三种药物对X射线损伤后肝实质细胞增殖的影响
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摘要
目的探讨三种药物对X射线损伤后肝实质细胞增殖的影响。方法将人肝实质HL-7702细胞株分为5组:异甘草酸镁组(照射前加入异甘草酸镁注射液)、还原型谷胱甘肽组(照射前加入注射用还原型谷胱甘肽)、多烯磷脂酰胆碱组(照射前加入多烯磷脂酰胆碱注射液)、对照组(单纯照射)和非照射组(正常细胞)。使用6 MV X射线照射细胞株,采用倒置显微镜观察各组照射24 h、48 h及72 h后的细胞形态,使用四甲基偶氮唑盐(MTT)法检测细胞存活率,分析多烯磷脂酰胆碱注射液、还原型谷胱甘肽注射液及异甘草酸镁注射液对受损肝细胞增殖修复的影响。结果倒置显微镜观察示非照射组细胞呈团状分布,分裂快,细胞为多角形,体积较小,对照组细胞生长受到显著抑制,细胞核变大,细胞边缘不清晰,体积变大且肿胀,细胞增殖变慢;细胞活力检测示照射24 h时肝细胞受抑制不显著,主要是细胞形态发生变化,但照射48 h及72 h时对照组细胞吸光度较非照射组显著降低,差异有统计学意义(P <0.05)。三种吸光度最高的药物浓度下,异甘草酸镁(1.0 mg/ml)对细胞增殖的有效率为38.60%,多烯磷脂酰胆碱(250 mol/L)为11.58%,GSH(0.1 mg/ml)未显示促增殖作用。照射24 h、48 h及72 h时,1.0 mg/ml异甘草酸镁组、250 mol/L多烯磷脂酰胆碱组的细胞A值均高于对照组及GSH组(P <0.05),24 h、48 h时1.0 mg/ml异甘草酸镁组和250 mol/L多烯磷脂酰胆碱组MDA含量低于对照组及GSH组,48 h时细胞SOD活力高于对照组及GSH组,差异均有统计学意义(P <0.05)。结论异甘草酸镁对X射线照射后人肝实质细胞增殖修复显著优于多烯磷脂酰胆碱及GSH,对放射性肝损伤治疗前景广阔,但具体作用机制仍需更深入研究。
Objective To investigate the effects of three drugs on the proliferation of hepatocytes after X-ray injury. Methods Human liver parenchyma HL-7702 cell lines were divided into 5 groups: magnesium isoglycyrrhizinate group(adding magnesium isoglycyrrhizinate injection before irradiation), reduced glutathione group(adding reduced glutathione before irradiation), polyene phosphatidylcholine group(adding polyene phosphatidylcholine before irradiation), control group(simple irradiation) and non-irradiated group(normal cells). The cells were irradiated with 6 MV X-raysmorphology and was observed by inverted microscope at 24 h, 48 h and 72 h after irradiation. The cell viability was measured by MTT assay. The effects of polyene phosphatidylcholine injection, reduced glutathione injection and magnesium isoglycyrrhizinate injection on proliferation and repairment of injured hepatocytes were analyzed. Results Inverted microscope observation showed that the non-irradiated group showed a cluster-like distribution, the division was fast, the cells were polygonal, and the volume was small. The growth of the control group was significantly inhibited, the nucleus became larger, the cell edge was unclear, the volume became larger and swollen, and the cell proliferation Slow down. The cell viability assay showed that the inhibition of hepatocytes was not significant at 24 h after irradiation, mainly due to the change of cell morphology. However, the absorbance of the control group was significantly lower than that of the non-irradiated group at 48 h and 72 h after irradiation(P < 0.05).The effects of magnesium isoglycyrrhizinate(1.0 mg/ml) and polyene phosphatidylcholine(250 mol/L) on cell proliferation were 38.60% and 11.58%, respectively. GSH(0.1 mg/ml) showed no effect of promoting proliferation. At 24 h, 48 h and 72 h after irradiation, the A values of magnesium isoglycyrrhizinate group(1.0 mg/ml) and polyene phosphatidylcholine group(250 mol/L) were higher than those of control group and GSH group. The content of MDA in magnesium isoglycyrrhizinate group(1.0 mg/ml) and polyene phosphatidylcholine group(250 mol/L) were lower than thoset in control group and GSH group at 24 h and 48 h after irradiation, and the cell SOD activity at 48 h was significantly higher than that in control group and GSH group(P < 0.05). Conclusions Magnesium glycyrrhizinate is superior to polyene phosphatidylcholine and GSH in the proliferation of human hepatocytes after X-ray irradiation. It has a broad prospect for the treatment of radiation-induced liver injury, but the specific mechanism still needs further study.
Objective To investigate the effects of three drugs on the proliferation of hepatocytes after X-ray injury. Methods Human liver parenchyma HL-7702 cell lines were divided into 5 groups: magnesium isoglycyrrhizinate group(adding magnesium isoglycyrrhizinate injection before irradiation), reduced glutathione group(adding reduced glutathione before irradiation), polyene phosphatidylcholine group(adding polyene phosphatidylcholine before irradiation), control group(simple irradiation) and non-irradiated group(normal cells). The cells were irradiated with 6 MV X-raysmorphology and was observed by inverted microscope at 24 h, 48 h and 72 h after irradiation. The cell viability was measured by MTT assay. The effects of polyene phosphatidylcholine injection, reduced glutathione injection and magnesium isoglycyrrhizinate injection on proliferation and repairment of injured hepatocytes were analyzed. Results Inverted microscope observation showed that the non-irradiated group showed a cluster-like distribution, the division was fast, the cells were polygonal, and the volume was small. The growth of the control group was significantly inhibited, the nucleus became larger, the cell edge was unclear, the volume became larger and swollen, and the cell proliferation Slow down. The cell viability assay showed that the inhibition of hepatocytes was not significant at 24 h after irradiation, mainly due to the change of cell morphology. However, the absorbance of the control group was significantly lower than that of the non-irradiated group at 48 h and 72 h after irradiation(P < 0.05).The effects of magnesium isoglycyrrhizinate(1.0 mg/ml) and polyene phosphatidylcholine(250 mol/L) on cell proliferation were 38.60% and 11.58%, respectively. GSH(0.1 mg/ml) showed no effect of promoting proliferation. At 24 h, 48 h and 72 h after irradiation, the A values of magnesium isoglycyrrhizinate group(1.0 mg/ml) and polyene phosphatidylcholine group(250 mol/L) were higher than those of control group and GSH group. The content of MDA in magnesium isoglycyrrhizinate group(1.0 mg/ml) and polyene phosphatidylcholine group(250 mol/L) were lower than thoset in control group and GSH group at 24 h and 48 h after irradiation, and the cell SOD activity at 48 h was significantly higher than that in control group and GSH group(P < 0.05). Conclusions Magnesium glycyrrhizinate is superior to polyene phosphatidylcholine and GSH in the proliferation of human hepatocytes after X-ray irradiation. It has a broad prospect for the treatment of radiation-induced liver injury, but the specific mechanism still needs further study.
引文
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[13]黄蓓,夏小健,孙起武,等.1例非小细胞肺癌患者药物性肝损伤的用药分析[J].中南药学,2017,15(10):1494-1498.
[14]陈昱延,周一凡,蒋大伟,等.干扰VPS4B表达对肝癌细胞增殖和细胞周期的影响[J].南通大学学报(医学版),2016,36(1):10-13.
[15]Petta S,Valenti L,Marra F,et al.MERTK rs4374383 polymorphism affects the severity of fibrosis in nonalcoholic fatty liver disease[J].JHepatol,2016,64(3):682-690.
[16]高世乐,胡宗涛,董六一,等.肝纤维化4项在原发性肝癌放疗前后变化的意义[J].医学研究杂志,2017,46(11):153-156.
[17]申刚,嘉红云,陈德基,等.miR-106b表达对人肝细胞肝癌细胞增殖能力的影响[J].中华肿瘤杂志,2014,36(7):489-495..
[18]杨光辉,薛涛,马博,等.门冬氨酸鸟氨酸对原发性肝癌放疗患者血管内皮生长因子的影响[J].中国医药导报,2014,11(15):65-67,71.
[19]许涛,景红霞,李林均,等.TACE、IMRT、HIFU联合DC-CIK治疗局部晚期原发性肝癌的效果[J].广东医学,2016,37(12):1846-1849.
[20]满晓波.肝细胞生长因子对肝细胞、肝癌细胞增殖、运动和细胞内游离钙离子浓度变化的影响[D].上海:第二军医大学,1997.
[21]嵇晓辉,范秉琳,张红鸽,等.MicroRNA-100对肝癌细胞增殖活力和细胞周期的影响[J].中国病理生理杂志,2013,29(1):108-111.
[22]刘子文,刘长征,刘卫,等.全反式维甲酸抑制肝癌细胞增殖的作用机制研究[J].中华肝胆外科杂志,2012,18(5):386-388.
[23]郭学军,曹传辉,孙景苑,等.miR-128a在肝细胞癌中表达上调并通过靶向调控RND3促进肝癌细胞增殖[J].南方医科大学学报,2014,34(10):1408-1413.
[24]陶颖,陈娟.沉默信息调节因子3对肝癌细胞增殖的影响[J].南方医科大学学报,2016,36(2):195-199.
[25]刘瑶,贺兴波,舒涛,等.miR-141表达异常对人肝癌细胞恶性生物学表型的影响[J].中国病理生理杂志,2016,32(2):215-220.
[2]张润萍,宁宇,刘娜,等.益气活血中药对放射性肝损伤的保护作用及对肝纤维化指标的影响[J].中国临床医生杂志,2017,45(10):112-114.
[3]Hayes CN,Chayama K.MicroRNAs as biomarkers for liver disease and hepatocellular carcinoma[J].Int J Mol Sci,2016,17(3):280.
[4]陈一兴,曾昭冲,孙菁,等.肝细胞肝癌经动脉化疗栓塞后行立体定向放疗的初步疗效观察[J].中国临床医学,2017,24(2):224-228.
[5]Qiu B,Wang J,Yu Y,et al.DJ-1 promotes development of DEN-induced hepatocellular carcinoma and proliferation of liver cancer cells[J].Oncotarget,2017,8(5):8499-8511.
[6]周蔼斌,周俊平.细胞因子及其信号通路在放射性肝损伤发病中的作用研究进展[J].实用肝脏病杂志,2016,19(6):766-769.
[7]Bilezik?i B,Haberal AN,Demirhan B.Hepatocyte growth factor in patients with three different stages of chronic liver disease including hepatocellular carcinoma,cirrhosis and chronic hepatitis:an immunohistochemical study[J].Can J Gastroenterol,2016,15(3):159-165.
[8]王琴,张东成,董超,等.三维适形放疗放射野布局对治疗原发性肝癌患者放射性肝损伤的影响[J].实用肝脏病杂志,2017,20(3):366-367.
[9]Wilhelm A,Aldridge V,Haldar D,et al.CD248/endosialin critically regulates hepatic stellate cell proliferation during chronic liver injury via a PDGF-regulated mechanism[J].Gut,2016,65(7):1175-1185.
[10]吕东来,陆林,卢虎生,等.三维适形放射治疗原发性肝癌患者发生放射性肝损伤相关因素分析[J].实用肝脏病杂志,2017,20(1):89-92.
[11]Crawford DR,Ilic Z,Guest I,et al.Characterization of liver injury,oval cell proliferation and cholangiocarcinogenesis in glutathione S-transferase A3 knockout mice[J].Carcinogenesis,2017,38(7):717-727.
[12]次旦旺久,畅智慧,赵相轩,等.125I放射性粒子持续照射抑制HepG2肝癌细胞增殖的作用机制[J].临床肝胆病杂志,2016,32(10):1920-1924.
[13]黄蓓,夏小健,孙起武,等.1例非小细胞肺癌患者药物性肝损伤的用药分析[J].中南药学,2017,15(10):1494-1498.
[14]陈昱延,周一凡,蒋大伟,等.干扰VPS4B表达对肝癌细胞增殖和细胞周期的影响[J].南通大学学报(医学版),2016,36(1):10-13.
[15]Petta S,Valenti L,Marra F,et al.MERTK rs4374383 polymorphism affects the severity of fibrosis in nonalcoholic fatty liver disease[J].JHepatol,2016,64(3):682-690.
[16]高世乐,胡宗涛,董六一,等.肝纤维化4项在原发性肝癌放疗前后变化的意义[J].医学研究杂志,2017,46(11):153-156.
[17]申刚,嘉红云,陈德基,等.miR-106b表达对人肝细胞肝癌细胞增殖能力的影响[J].中华肿瘤杂志,2014,36(7):489-495..
[18]杨光辉,薛涛,马博,等.门冬氨酸鸟氨酸对原发性肝癌放疗患者血管内皮生长因子的影响[J].中国医药导报,2014,11(15):65-67,71.
[19]许涛,景红霞,李林均,等.TACE、IMRT、HIFU联合DC-CIK治疗局部晚期原发性肝癌的效果[J].广东医学,2016,37(12):1846-1849.
[20]满晓波.肝细胞生长因子对肝细胞、肝癌细胞增殖、运动和细胞内游离钙离子浓度变化的影响[D].上海:第二军医大学,1997.
[21]嵇晓辉,范秉琳,张红鸽,等.MicroRNA-100对肝癌细胞增殖活力和细胞周期的影响[J].中国病理生理杂志,2013,29(1):108-111.
[22]刘子文,刘长征,刘卫,等.全反式维甲酸抑制肝癌细胞增殖的作用机制研究[J].中华肝胆外科杂志,2012,18(5):386-388.
[23]郭学军,曹传辉,孙景苑,等.miR-128a在肝细胞癌中表达上调并通过靶向调控RND3促进肝癌细胞增殖[J].南方医科大学学报,2014,34(10):1408-1413.
[24]陶颖,陈娟.沉默信息调节因子3对肝癌细胞增殖的影响[J].南方医科大学学报,2016,36(2):195-199.
[25]刘瑶,贺兴波,舒涛,等.miR-141表达异常对人肝癌细胞恶性生物学表型的影响[J].中国病理生理杂志,2016,32(2):215-220.