柿树寄生植物保肝作用活性部分的筛选及其对小鼠急性肝损伤的影响
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摘要
目的:通过建立四氯化碳(CCl_4)急性肝损伤模型筛选出柿树寄生植物具有保肝作用的活性部分,并探讨其对肝损伤小鼠肝细胞的保护作用。方法:(1)制备柿树寄生植物石油醚萃取物、乙酸乙酯萃取物、正丁醇萃取物,建立CCl_4急性肝损伤模型,通过检测各萃取物处理组小鼠血清谷丙转氨酶(ALT)、谷草转氨酶(AST)水平及肝组织丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性,筛选柿树寄生植物的活性部分。(2)将60只昆明小鼠随机分为正常组、模型组、联苯双酯(150 mg/kg)组及正丁醇萃取物低剂量(11.67 mg/kg)组、中剂量(23.34 mg/kg)组、高剂量组(46.67 mg/kg)组,灌胃给药8 d后,除正常组外,其他各组均建立急性肝损伤模型,检测各组小鼠血清白细胞介素(IL)-6和IL-1β水平,并采用免疫组织化学染色法检测肝组织IL-6蛋白表达量。结果:柿树寄生植物各萃取物处理组小鼠肝脏、脾脏、胸腺系数比较,差异均无统计学意义(均P>0.05);与模型组比较,正丁醇萃取物组肝组织MDA含量及血清AST、ALT水平明显降低,肝组织SOD活性明显提高(P<0.05),而石油醚萃取物组和乙酸乙酯萃取物组各指标比较,差异均无统计学意义(均P>0.05)。正丁醇萃取物中、高剂量组小鼠血清IL-6水平及肝组织IL-6蛋白表达量均较模型组显著降低,正丁醇萃取物各剂量组血清IL-1β水平均显著降低(P<0.05),且中、高剂量组IL-6水平低于低剂量组(P<0.05),而IL-1β水平在各剂量组之间无明显差异(P>0.05)。结论:柿树寄生植物正丁醇萃取物是柿树寄生植物保肝作用的活性部分,能明显减轻CCl_4所致的肝损伤,其机制与抑制炎症因子表达,提高机体抗氧化能力,降低氧化应激有关。
Objective:To observe the protective effect of parasitic active part of persimmon on acute liver injury(ALI) induced by carbon tetrachloride(CCl_4) in mice.Methods:(1) The petroleum ether extract,ethyl acetate extract,and n-butanol extract from persimmon were prepared.ALI model of mice was induced by CCl_4,which was used to screen the parasitic active part of persimmon.(2) A total of 60 Kunming mice were divided into control group,model group,bidentate group,and low-,medium-and high-dose(11.67,23.34 and 46.67 mg/kg) of n-butanol extract groups.The mice were treated with 150 mg/kg bidentate or different doses of n-butanol extract via gavage for 8 days,and then the ALI model was established.The levels of serum interleukin(IL)-6 and IL-1β were measured.The expression of IL-6 protein in liver tissues was detected by immunohistochemical staining.Results:There were no significant differences in the ratios of liver,spleen and thymic weight to the body weight among different extracts groups(P>0.05).Compared with the model group,the MDA content in liver tissues and the levels of serum AST and ALT in the n-butanol extract group were significantly decreased,while the SOD activity in liver tissues was significantly increased(P<0.05),while no obvious difference was noted between the petroleum ether extract group and ethyl acetate extract group(P>0.05).The serum IL-6 level and IL-6 protein expression level in liver tissues in medium-and high-dose of n-butanol extract groups were lower than those in the model group,and the serum IL-1β level in low-,medium-and high-dose of n-butanol extract groups was significantly lower(P<0.05).Moreover,the serum level of IL-6 in medium-and high-dose of n-butanol extract groups was lower than that in low-dose group(P<0.05),but there was no significant difference in the serum IL-1β concentration among different doses groups(P<0.05).Conclusion:N-butanol extract was an active part of persimmon,which could protect mice against CCl_4-induced ALI by suppressing inflammation and oxidative stress.
Objective:To observe the protective effect of parasitic active part of persimmon on acute liver injury(ALI) induced by carbon tetrachloride(CCl_4) in mice.Methods:(1) The petroleum ether extract,ethyl acetate extract,and n-butanol extract from persimmon were prepared.ALI model of mice was induced by CCl_4,which was used to screen the parasitic active part of persimmon.(2) A total of 60 Kunming mice were divided into control group,model group,bidentate group,and low-,medium-and high-dose(11.67,23.34 and 46.67 mg/kg) of n-butanol extract groups.The mice were treated with 150 mg/kg bidentate or different doses of n-butanol extract via gavage for 8 days,and then the ALI model was established.The levels of serum interleukin(IL)-6 and IL-1β were measured.The expression of IL-6 protein in liver tissues was detected by immunohistochemical staining.Results:There were no significant differences in the ratios of liver,spleen and thymic weight to the body weight among different extracts groups(P>0.05).Compared with the model group,the MDA content in liver tissues and the levels of serum AST and ALT in the n-butanol extract group were significantly decreased,while the SOD activity in liver tissues was significantly increased(P<0.05),while no obvious difference was noted between the petroleum ether extract group and ethyl acetate extract group(P>0.05).The serum IL-6 level and IL-6 protein expression level in liver tissues in medium-and high-dose of n-butanol extract groups were lower than those in the model group,and the serum IL-1β level in low-,medium-and high-dose of n-butanol extract groups was significantly lower(P<0.05).Moreover,the serum level of IL-6 in medium-and high-dose of n-butanol extract groups was lower than that in low-dose group(P<0.05),but there was no significant difference in the serum IL-1β concentration among different doses groups(P<0.05).Conclusion:N-butanol extract was an active part of persimmon,which could protect mice against CCl_4-induced ALI by suppressing inflammation and oxidative stress.
引文
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[2] SAKALLI CETIN E,TETIKER H,IIHAN CELIK O,et al.Methotrexate-induced nephrotoxicity in rats:protective effect of mistletoe (Viscum album L.) extract [J].Complement Med Res,2017,24(6):364-370.
[3] RANJBAR A,SHARIFZADEH M,KARIMI K,et al.Propofol attenuates toxic oxidative stress by CCl4 in liver mitochondria and blood in rat[J].ranian Journal of Pharmaceutical Research,2014,13(1):253-262.
[4] MISTRY S,DUTT K R,JENA J.Protective effect of Sida cordata leaf extract against CCl 4 induced acute liver toxicity in rats[J].Asian Pac J Trop Med,2013,6(4):280-284.
[5] WEBER L W,BOLL M,STAMPFL A.Hepatotoxicity and mechanism of action of haloalkanes:Carbon tetrachloride as a toxicological model[J].Critical Reviews in Toxicology,2003,33(2):105-136.
[6] 尹连红,许有威.金樱子胶囊对四氯化碳诱导小鼠急性肝损伤的保护作用研究[J].大连医科大学学报,2018,40(1):27-32.
[7] 郑作文,李以军,张文涛.壮药依肝达对小鼠急性肝损伤的保护作用[J].中成药,2015,37(11):2518-2520.
[8] YANG B Y,ZHANG X Y,GUAN S W,et al.Protective effect of procyanidin B2 against CCl4-induced acute liver injury in mice[J].Molecules,2015,20(7):12250-12265.
[9] LI S Q,ZHU S,HAN H M,et al.IL-6 trans-signaling plays important protective roles in acute liver injury induced by acetaminophen in mice[J].J Biochem Mol Toxicol,2015,29(6):288-297.
[10] LIU T,XIA Y,LI J,et al.Shikonin attenuates concanavalin A-induced acute liver injury in mice via inhibition of the JNK pathway[J].Mediators Inflamm,2016,27(3):156-178.
[11] LI L.Preventive effects of interleukin-6 in lipopolysaccharide/D-galactosamine induced acute liver injury via regulating inflammatory response in hepatic macrophages[J].Int Immunopharmacol,2017(51):99-106.