慈姑多糖对异烟肼和利福平联用致损伤小鼠肝细胞凋亡的保护作用
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摘要
目的探讨慈姑多糖对异烟肼和利福平联用致损伤小鼠肝细胞凋亡基因Bax、Bcl-2表达的影响。方法制造异烟肼(INH)和利福平(RFP)联用小鼠肝损伤模型,HE染色观察肝组织病理变化,筛选慈姑多糖肝损伤体内保护最佳剂量。RT-PCR和Western-blot检测小鼠肝细胞中Bax和Bcl-2的mRNA及蛋白表达。结果与对照组相比,INH/RFP模型组小鼠肝组织形态明显改变,小鼠肝细胞中Bax、Bcl-2的表达上升,Bcl-2/Bax减小(P<0.05);与模型组相比,慈姑多糖各剂量均能减轻INH/RFP所致小鼠肝组织病变,高剂量组改善最明显,作为最佳剂量进行后续实验;小鼠肝细胞中高剂量慈姑多糖组Bax表达下降,Bcl-2的表达上升,Bcl-2/Bax增大(P<0.05)。结论慈姑多糖可以对抗INH/RFP联用致肝损伤,可能与抑制Bax的表达并提高Bcl-2的表达从而减少细胞的凋亡有关。
Objective To explore the effects of sagittaria sagittifolin polysaccharide(SSP) on the expression of Bax and Bcl-2 in isoniazid(INH)/rifampicin(RfP)-induced hepatic injury liver cells in mice.Methods Models of live injured mice were induced with INH/RFP.Liver tissues pathology was observed by using HE staining to determine the optimal SSP dose.RT-PCR and Western-blot assay were used to measure the levels of mRNA and protein of Bax and Bcl-2.Results Compared with the control group,liver tissues of mice from the model group presented with significant changes.The expression of Bcl-2 significantly increased in mice,and Bcl-2/Bax decreased(P<0.05).Compared with model group,pathological changes of liver tissues were alleviated in all SSP treated groups,especially in high-dose group.Expression of Bax decreased,expression of Bcl-2 significantly increased and Bcl-2/Bax increased in mouse liver cells(P<0.05).Conclusion SSP may protect liver cells against INH/RFP induced apoptosis by inhibiting the expression of Bax as well as increasing the expression of Bcl-2.Key
Objective To explore the effects of sagittaria sagittifolin polysaccharide(SSP) on the expression of Bax and Bcl-2 in isoniazid(INH)/rifampicin(RfP)-induced hepatic injury liver cells in mice.Methods Models of live injured mice were induced with INH/RFP.Liver tissues pathology was observed by using HE staining to determine the optimal SSP dose.RT-PCR and Western-blot assay were used to measure the levels of mRNA and protein of Bax and Bcl-2.Results Compared with the control group,liver tissues of mice from the model group presented with significant changes.The expression of Bcl-2 significantly increased in mice,and Bcl-2/Bax decreased(P<0.05).Compared with model group,pathological changes of liver tissues were alleviated in all SSP treated groups,especially in high-dose group.Expression of Bax decreased,expression of Bcl-2 significantly increased and Bcl-2/Bax increased in mouse liver cells(P<0.05).Conclusion SSP may protect liver cells against INH/RFP induced apoptosis by inhibiting the expression of Bax as well as increasing the expression of Bcl-2.Key
引文
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[13] 卢春凤,陈廷玉,赵锦程,等.槲皮素对INH诱导的大鼠肝损伤及肝组织Bcl-2和Bax表达的影响[J].解剖学研究,2010,32(1):28-31.Lu CF, Chen TY, Zhao JC, et al. Effect of quercetin on INH injuried liver and expressions of Bcl-2 and Bax in liv-er[J]. Anatomy Research, 2010,32(1):28-31.
[14] Chen X, Xu J, Zhang C, et al. The protective effects of ursodeoxycholic acid on isoniazid plus rifampicin induced liver injury in mice[J]. Eur. J. Pharmacol, 2011,659(1):53-60.
[15] Cao P, Sun JL, Sullivan M A, et al. Angelica sinensis polysaccharide protects against acetaminophen-induced acute liver injury and cell death by suppressing oxidative stress and hepatic apoptosis in vivo and in vitro[J]. Int. J. Biol. Macromol, 2018,111:1133-1139.
[16] Wang X, Liu JR, Zhang XH, et al. Seabuckthorn berry polysaccharide extracts protect against acetaminophen induced hepatotoxicity in mice via activating the Nrf-2/HO-1-SOD-2 signaling pathway[J]. Phytomedicine, 2018,38:90-97.
[17] Dong YZ, Huang JC, Lin X, et al. Hepatoprotective effects of Yulangsan polysaccharide against isoniazid and rifampicin-induced liver injury in mice[J]. J Ethnopharmacol, 2014,152(1):201-206.
[18] 罗永会,张翠香,徐春萍.慈姑多糖降血糖的生物活性研究[J].大理学院学报,2012,11(12):4-5,9.Luo YH, Zhang CX, Xu CP, et al. Research on hypoglycemic effect of the sagittaria sagittifolin polysaccharide[J]. Journal of Dali University, 2012,11(12):4-5,9.
[19] 欧丽兰,余昕,张椿,等.慈姑多糖的提取工艺及其抗肿瘤活性[J].中成药,2016,38(8):1835-1838.Ou LL, Yu X, Zhang C, et al. Antitumor activity of sagittaria sagittifolin polysaccharide[J]. Chinese Traditional Patent Medicine, 2016,38(8):1835-1838.
[2] Cao J, Mi YJ, Shi CL, et al. First-line anti-tuberculosis drugs induce hepatotoxicity: a novel mechanism based on a urinary metabolomics platform[J]. Biochem. Biophys. Res. Commun., 2018,497(2):485-491.
[3] Ou LL, Shui PX, Zhu Y, et al. Study on extraction of polysaccharides and antioxidant activity of sagittaria sagittifolia L. polysaccharide[J]. Agricultural Science & Technology, 2017,18(4):724-732.
[4] 廖艳,孙奇,彭桂英,等.慈菇对异烟肼和利福平致大鼠肝损伤的保护作用[J].北京中医药大学学报,2012,35(7):466-469.Liao Y, Sun Q, Peng GY, et al. Protective effect of Cigu (Sagittaria Sagittifolia) on liver damaged by isoniazid and rifampin in rats[J]. Journal of Beijing University of Traditional Chinese Medicine, 2012,35(7):466-469.
[5] 李冰,王春国,王晶,等.慈姑水提部位对异烟肼和利福平合用致小鼠肝损伤的影响[J].吉林中医药,2016,36(8):818-821.Li B, Wang CG, Wang J, et al. Water extract from sagittaria sagittifolia on hepatic injury induced by co-administration of isoniazid and rifampicin[J]. Journal of Jilin Traditional Chinese Medicine, 2016,36(8):818-821.
[6] 廖艳,李冰,林殷,等.慈姑多糖和慈姑水提部位对异烟肼和利福平合用致肝细胞损伤保护作用的比较研究[J].北京中医药,2017,36(5):415-419.Liao Y, Li B, Lin Y, et al. Comparative study on the protective effect of polysaccharides and aqueous extracts of sagittaria sagittifolia against hepatocellular injury caused by co-administration of isoniazid and rifampin[J]. Journal of Beijing Traditional Chinese Medicine, 2017,36(5):415-419.
[7] Wang J, Luo WZ, Li B, et al. Sagittaria sagittifolia polysaccharide protects against isoniazid- and rifampicin-induced hepatic injury via activation of nuclear factor E2-related factor 2 signaling in mice[J]. J Ethnopharmacol, 2018,227:237-245.
[8] Ke XH, Wang CG, Luo WZ, et al. Metabolomic study to determine the mechanism underlying the effects of polysaccharide on isoniazid- and rifampicin-induced hepatotoxicity in mice[J]. Molecules, 2018,23:1-11.
[9] Wang C, Fan RQ, Zhang YX, et al. Naringenin protects against isoniazid-and rifampicin-induced apoptosis in hepatic injury[J]. World J. Gastroenterol, 2016,22(44):9775-9783.
[10] 王晶,王春国,李冰,等.苯酚-硫酸法测定慈姑中多糖的含量[J].吉林中医药,2017,37(12):1258-1260.Wang J, Wang CG, Li B, et al. Determination of polysaccharides from sagittaria by method of H2SO4-phenol[J]. Journal of Jilin Traditional Chinese Medicine, 2017,37(12):1258-1260.
[11] Wang P, Pradhan K, Zhong X, et al. Isoniazid metabolism and hepatotoxicity[J]. Acta Pharmaceutica Sinica B, 2016,6(5):384-392.
[12] Chowdhury A, Santra A, Bhattacharjee K, et al. Mitochondrial oxidative stress and permeability transition in isoniazid and rifampicin induced liver injury in mice[J]. J. Hepatol, 2006,45(1):117-126.
[13] 卢春凤,陈廷玉,赵锦程,等.槲皮素对INH诱导的大鼠肝损伤及肝组织Bcl-2和Bax表达的影响[J].解剖学研究,2010,32(1):28-31.Lu CF, Chen TY, Zhao JC, et al. Effect of quercetin on INH injuried liver and expressions of Bcl-2 and Bax in liv-er[J]. Anatomy Research, 2010,32(1):28-31.
[14] Chen X, Xu J, Zhang C, et al. The protective effects of ursodeoxycholic acid on isoniazid plus rifampicin induced liver injury in mice[J]. Eur. J. Pharmacol, 2011,659(1):53-60.
[15] Cao P, Sun JL, Sullivan M A, et al. Angelica sinensis polysaccharide protects against acetaminophen-induced acute liver injury and cell death by suppressing oxidative stress and hepatic apoptosis in vivo and in vitro[J]. Int. J. Biol. Macromol, 2018,111:1133-1139.
[16] Wang X, Liu JR, Zhang XH, et al. Seabuckthorn berry polysaccharide extracts protect against acetaminophen induced hepatotoxicity in mice via activating the Nrf-2/HO-1-SOD-2 signaling pathway[J]. Phytomedicine, 2018,38:90-97.
[17] Dong YZ, Huang JC, Lin X, et al. Hepatoprotective effects of Yulangsan polysaccharide against isoniazid and rifampicin-induced liver injury in mice[J]. J Ethnopharmacol, 2014,152(1):201-206.
[18] 罗永会,张翠香,徐春萍.慈姑多糖降血糖的生物活性研究[J].大理学院学报,2012,11(12):4-5,9.Luo YH, Zhang CX, Xu CP, et al. Research on hypoglycemic effect of the sagittaria sagittifolin polysaccharide[J]. Journal of Dali University, 2012,11(12):4-5,9.
[19] 欧丽兰,余昕,张椿,等.慈姑多糖的提取工艺及其抗肿瘤活性[J].中成药,2016,38(8):1835-1838.Ou LL, Yu X, Zhang C, et al. Antitumor activity of sagittaria sagittifolin polysaccharide[J]. Chinese Traditional Patent Medicine, 2016,38(8):1835-1838.