铁皮石斛粗多糖对刀豆蛋白A诱导的小鼠急性肝损伤的保护作用及机制研究
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摘要
目的:观察铁皮石斛粗多糖(Dendrobium Officinale,DO)对刀豆蛋白A(concanavalin A,ConA)诱导的小鼠急性肝损伤的保护作用并探讨相关机制。方法:利用ConA(20 mg/kg)制备小鼠急性肝损伤模型,分别给予300、600、1200 mg/kg剂量的铁皮石斛多糖(DOPS)连续灌胃20天。全自动生化分析仪检测血清ALT、AST、Alb、TP及A/G比; ELISA试剂盒检测TNF-α、INF-γ;肝组织HE染色进行肝组织病理学损伤分级评价;试剂盒检测SOD、GSH、MDA水平;TUNEL染色法检测肝细胞凋亡情况;Western blot检测Nrf2、HO-1、p-JNK、BCl-2的蛋白表达情况。结果:与ConA诱导的小鼠急性肝损伤模型组相比,铁皮石斛组小鼠血清ALT、 AST、TNF-α、INF-γ值明显降低,Alb、TP及A/G比明显升高;肝组织病理损伤分级明显降低;肝组织的SOD、GSH水平明显升高,MDA水平明显降低。与模型组相比,铁皮石斛组小鼠肝细胞凋亡明显减少;Nrf2、HO-1、BCl-2的蛋白表达明显升高,p-JNK明显降低。结论:本研究表明铁皮石斛对ConA诱导的小鼠急性肝损伤具有保护作用,其机制可能与铁皮石斛上调Nrf2信号通路,抑制氧化应激,进而抑制JNK信号通路相关。
Objective: To investigate the protective effect and the mechanism of Dendrobium Officinale( DO) on concanavalin A( ConA)-induced acute liver injury in mice Methods: Acute liver injury mice model was established by ConA( 20 mg/kg). Mice were orally administered with300、600、1200 mg/kg of DOPS extract for 20 days,respectively. The serum levels of ALT,AST,Alb and TP were determined by automatic biochemical analyzer and A/G ratio was calculated. Levels of TNF-α and IFN-γ were assayed by ELISA. Liver tissues were stained by HE and the histopathological changes were evaluated. The contents of SOD,GSH,MDA were assayed by related kits. Apoptosis of hepatic cells was detected by TUNEL staining. The protein expressions of Nrf2,HO-1,p-JNK and BCl-2 were detected by Western blot. Results: Compared with the model group,the serum levels of ALT,AST,TNF-、INF-γ treated with DO were significantly decreased and Alb,TP,A/G were significantly increased. The histopathological grade of liver damage decreased. The levels of SOD,GSH in liver tissues were significantly increased and MDA level was decreased. Compared with the model group,the hepatic cell apoptosis of DO groups were significantly decreased. The protein expressions levels of Nrf2,HO-1 and BCl-2 were significantly increased and p-JNK was significantly decreased. Conclusion: The results show that DO has the protective effect on ConA-induced acute liver injury mice. Its mechanism may be related to up-regulating Nrf2 signaling pathway,inhibiting oxidative stress and JNK signaling pathway.
Objective: To investigate the protective effect and the mechanism of Dendrobium Officinale( DO) on concanavalin A( ConA)-induced acute liver injury in mice Methods: Acute liver injury mice model was established by ConA( 20 mg/kg). Mice were orally administered with300、600、1200 mg/kg of DOPS extract for 20 days,respectively. The serum levels of ALT,AST,Alb and TP were determined by automatic biochemical analyzer and A/G ratio was calculated. Levels of TNF-α and IFN-γ were assayed by ELISA. Liver tissues were stained by HE and the histopathological changes were evaluated. The contents of SOD,GSH,MDA were assayed by related kits. Apoptosis of hepatic cells was detected by TUNEL staining. The protein expressions of Nrf2,HO-1,p-JNK and BCl-2 were detected by Western blot. Results: Compared with the model group,the serum levels of ALT,AST,TNF-、INF-γ treated with DO were significantly decreased and Alb,TP,A/G were significantly increased. The histopathological grade of liver damage decreased. The levels of SOD,GSH in liver tissues were significantly increased and MDA level was decreased. Compared with the model group,the hepatic cell apoptosis of DO groups were significantly decreased. The protein expressions levels of Nrf2,HO-1 and BCl-2 were significantly increased and p-JNK was significantly decreased. Conclusion: The results show that DO has the protective effect on ConA-induced acute liver injury mice. Its mechanism may be related to up-regulating Nrf2 signaling pathway,inhibiting oxidative stress and JNK signaling pathway.
引文
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[10]宋美芳,李光,陈曦,等.两种石斛多糖提高小鼠免疫活性的初步研究.中国药学杂志,2013,48(6)∶428～431.
[11]邱现创,赵宁,李晨,等.铁皮石斛多糖提取工艺优化及对果蝇抗氧化能力的影响.食品科学,2018,39(2)∶ 273～280.
[12]钟惠娟,陈璐,周洁,等.铁皮石斛多糖对高糖诱导的血管内皮依赖性舒张功能的影响.中国新药杂志, 2017,26(12)∶ 1443～1449.
[13]吴人照,杨兵勋,李亚平,等.铁皮石斛多糖对SHR-sp大鼠抗高血压中风作用的实验研究.中国中医药科技, 2011,18(3)∶ 204～210.
[14]王杰,葛颖华,周萃,等.鲜铁皮石斛提取物抗Lewis肺癌的机制研究.中国现代应用药学,2014,31(8)∶953～957.
[15]陈健,戚辉,李金标,等.铁皮石斛多糖促进毛发生长的实验研究.中国中药杂志,2014,39(2)∶291～295.
[16]王玲,唐德强,王佳佳,等.铁皮石斛原球茎与野生铁皮石斛多糖的抗菌及体外抗氧化活性比较.西北农林科技大学学报(自然科学版),2016,44(6)∶ 167～180.
[17]林为艺,颜美秋,吕圭源,等.铁皮石斛茎非多糖与粗多糖体内外抗氧化活性的比较.中药药理与临床, 2016,32(2)∶ 138～141.
[18]粱楚燕,李焕彬,侯少贞,等.铁皮石斛护肝及抗胃溃疡作用研究.世界科学技术-中医药现代化·中药研究,2013,15(2)∶ 233～236.
[19]汤小华,陈素红,吕圭源,等.铁皮石斛对小鼠急性酒精性肝损伤模型SOD、MDA、GSH-Px的影响. 浙江中医杂志, 2010,45(5)∶369～370.
[20]袁慧琦,梁楚燕,梁健,等.铁皮石斛对小鼠急性酒精性肝损伤的保护作用.暨南大学学报(自然科学与医学版),2016,37(5)∶ 384～388.
[21]王凯,眭丹娟,王长锁,等.5种石斛对四氯化碳致小鼠急性肝损伤的保护作用.中国中药杂志,2017,42(10)∶ 1945～1950.
[22]Wang F, Xue Y, Yang J Y, et al. Hepatoprotective effect of apple polyphenols against concanavalin A-induced immunological liver injury in mice. Chem Biol Interact, 2016,258∶ 159～165.
[23]Wang K P, Song Z Z, Wang H J, et al. Angelica sinensis polysaccharide attenuates concanavalin A-induced liver injury in mice. Int Immunopharmacol, 2016,31∶ 140～148.
[24]Chen K, Li J J, Li S N, et al. 15d-PGJ2 alleviates ConA-induced acute liver injury in mice by up-regulating HO-1 and reducing hepatic cell autophagy. Biomed Pharmacother, 2016, 80∶ 183～192.
[25]Liang J, Chen S X, Hu Y D. Protective roles and mechanisms of Dendrobium offcinal polysaccharides on secondary liver injury in acute colitis. Int J Biol Macromol, 2018, 107∶ 2201～2210.
[26]Ma J Q, Ding J, Zhang L, et al. Hepatoprotective properties of sesamin against CCl4 induced oxidative stress-mediated apoptosis in mice via JNK pathway. Food Chem Toxicol,2014,64∶ 41～48.
[27]刘乐江,唐圣松. ROS介导JNK信号通路的研究进展.现代生物医学进展,2010,10(7)∶1378～1380.
[2]张引强,唐旭东,郭朋,等. 荣肝合剂对刀豆蛋白A诱导急性免疫性肝损伤小鼠T淋巴细胞亚群的影响. 中西医结合肝病杂志, 2016,26(1)∶26～29.
[3]Tiegs G. Cellular and cytokine-mediated mechanisms of inflammation and its modulation in immune-mediated liver injury. Z Gastroenterol, 2007,45(1)∶ 63～70.
[4]Li J J, Xia Y J, Liu T, et al. Protective effects of astaxanthin on ConA-induced autoimmune hepatitis by the JNK/p-JNK pathway-mediated inhibition of autophagy and apoptosis. PLoS One, 2015,10(3)∶ 1～21.
[5]汪劼,易静. JNK介导的信号转导途径以及活性氧在其中的作用.生命科学,2006,18(4)∶ 361～367.
[6]Zhang Y D, Chen F. Reactive Oxygen Species(ROS), Troublemakers between Nuclear Factor- B (NF- B) and c-Jun NH2-terminal Kinase (JNK). Cancer Res, 2004,64(6)∶ 1902～1905.
[7]Okada K, Shoda J, Taguchi K, et al. Nrf2 counteracts cholestatic liver injury via stimulation of hepatic defense systems. Biochem Biophys Res Commun, 2009,389(3)∶ 431～436.
[8]Shin S M, Yang J H, Ki S H. Role of the Nrf2-ARE pathway in liver diseases. Oxid Med Cell Longev, 2013,2013∶ 763257.
[9]吕圭源,颜美秋,陈素红.铁皮石斛功效相关药理作用研究进展.中国中药杂志,2013,38(4)∶489～493.
[10]宋美芳,李光,陈曦,等.两种石斛多糖提高小鼠免疫活性的初步研究.中国药学杂志,2013,48(6)∶428～431.
[11]邱现创,赵宁,李晨,等.铁皮石斛多糖提取工艺优化及对果蝇抗氧化能力的影响.食品科学,2018,39(2)∶ 273～280.
[12]钟惠娟,陈璐,周洁,等.铁皮石斛多糖对高糖诱导的血管内皮依赖性舒张功能的影响.中国新药杂志, 2017,26(12)∶ 1443～1449.
[13]吴人照,杨兵勋,李亚平,等.铁皮石斛多糖对SHR-sp大鼠抗高血压中风作用的实验研究.中国中医药科技, 2011,18(3)∶ 204～210.
[14]王杰,葛颖华,周萃,等.鲜铁皮石斛提取物抗Lewis肺癌的机制研究.中国现代应用药学,2014,31(8)∶953～957.
[15]陈健,戚辉,李金标,等.铁皮石斛多糖促进毛发生长的实验研究.中国中药杂志,2014,39(2)∶291～295.
[16]王玲,唐德强,王佳佳,等.铁皮石斛原球茎与野生铁皮石斛多糖的抗菌及体外抗氧化活性比较.西北农林科技大学学报(自然科学版),2016,44(6)∶ 167～180.
[17]林为艺,颜美秋,吕圭源,等.铁皮石斛茎非多糖与粗多糖体内外抗氧化活性的比较.中药药理与临床, 2016,32(2)∶ 138～141.
[18]粱楚燕,李焕彬,侯少贞,等.铁皮石斛护肝及抗胃溃疡作用研究.世界科学技术-中医药现代化·中药研究,2013,15(2)∶ 233～236.
[19]汤小华,陈素红,吕圭源,等.铁皮石斛对小鼠急性酒精性肝损伤模型SOD、MDA、GSH-Px的影响. 浙江中医杂志, 2010,45(5)∶369～370.
[20]袁慧琦,梁楚燕,梁健,等.铁皮石斛对小鼠急性酒精性肝损伤的保护作用.暨南大学学报(自然科学与医学版),2016,37(5)∶ 384～388.
[21]王凯,眭丹娟,王长锁,等.5种石斛对四氯化碳致小鼠急性肝损伤的保护作用.中国中药杂志,2017,42(10)∶ 1945～1950.
[22]Wang F, Xue Y, Yang J Y, et al. Hepatoprotective effect of apple polyphenols against concanavalin A-induced immunological liver injury in mice. Chem Biol Interact, 2016,258∶ 159～165.
[23]Wang K P, Song Z Z, Wang H J, et al. Angelica sinensis polysaccharide attenuates concanavalin A-induced liver injury in mice. Int Immunopharmacol, 2016,31∶ 140～148.
[24]Chen K, Li J J, Li S N, et al. 15d-PGJ2 alleviates ConA-induced acute liver injury in mice by up-regulating HO-1 and reducing hepatic cell autophagy. Biomed Pharmacother, 2016, 80∶ 183～192.
[25]Liang J, Chen S X, Hu Y D. Protective roles and mechanisms of Dendrobium offcinal polysaccharides on secondary liver injury in acute colitis. Int J Biol Macromol, 2018, 107∶ 2201～2210.
[26]Ma J Q, Ding J, Zhang L, et al. Hepatoprotective properties of sesamin against CCl4 induced oxidative stress-mediated apoptosis in mice via JNK pathway. Food Chem Toxicol,2014,64∶ 41～48.
[27]刘乐江,唐圣松. ROS介导JNK信号通路的研究进展.现代生物医学进展,2010,10(7)∶1378～1380.