睡莲花烟花苷对半乳糖胺致小鼠急性肝损伤的保护作用

Protective effect of nicotiflorin from Nymphaea candida on D-Galinduced acute liver injury in mice

目的:研究睡莲花烟花苷对D-半乳糖胺(D-Gal)所致小鼠急性肝损伤的保护作用。方法:将60只昆明小鼠随机分为正常组,模型组,阳性对照联苯双酯组,烟花苷(25、50和100 mg/kg)给药组,共6组,每组10只。阳性对照联苯双酯组和烟花苷给药组按10 m L/kg每日灌胃给药1次,正常组和模型组每日灌胃等体积蒸馏水,连续灌胃7 d,末次给药1 h后,正常组小鼠腹腔注射生理盐水0.02 m L/g,其他各组腹腔注射D-Gal(800 mg/kg,按0.02 m L/g)造模。造模8 h后摘眼球取血,测定血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)、白细胞介素IL-1β(IL-1β)、白细胞介素(IL-6)、肿瘤坏死因子α(TNF-α)和干扰素γ(INF-γ)水平。颈椎脱臼处死小鼠,摘取肝脏、脾脏称取质量,计算脏器系数,取0.3 g肝组织制备匀浆,检测肝匀浆中丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH)及一氧化氮(NO)含量。并取肝左叶距边缘0.5 cm处小块组织,进行肝组织病理学检查。结果:与模型组比较,烟花苷在(50、100 mg/kg)剂量时,小鼠血清ALT和AST水平均显著降低(P均<0.01),血清中促炎因子IL-1、IL-6、TNF-α和INF-γ水平降低(P<0.05);肝脏、脾脏系数降低(P<0.05),肝匀浆中SOD和GSH活力均显著升高(P均<0.05),MDA和NO含量均显著降低(P均<0.05);肝病理检查结果显示与模型组比较,烟花苷各剂量组肝细胞损伤程度明显减轻,炎性细胞浸润显著减少,其中烟花苷50和100 mg/kg组较25 mg/kg组受损程度较轻、修复情况较好。结论:烟花苷对D-Gal致小鼠急性肝损伤具有保护作用,其作用与促进抗氧化酶、抑制促炎因子的表达有关。

OBJECTIVE:To study protective effect of nicotiflorin from Nymphaea candida on D-Gal-induced liverinjury in mice.METHODS:Sixty healthy adult mice were randomly divided into six groups:control,model,dimethyldiphenyl bicarboxylate(DDB,150mg/kg)and nicotiflorin(25,50,100mg/kg)groups.Chemicals were intragastricallyadministrated f or s even d ays.O ne h our a fter t he l ast a dministration,mice i n a ll e xcept t he c ontrol g roup w ere i ntraperitoneallyinjection w ith D-Gal(800m g/kg,0.02m L/g).A ll m ice w ere s acrificed a t t he8th h our,blood w as c ollected f rom t he e yeball,and serum ALT,AST,IL-1β,IL-6,TNF-αand INF-γwere determined.Mice were killed cervical dislocation,thequality o f t he b ody a fter t he a utopsy,the l iver,spleen,the q uality o f t he o rgan c oefficient,the l iver h omogenate w ere p lacedin-20freezer.The left hepatic lobes at0.5cm from the edge of tissues were fixed with10%formalin,dehydrated,transparentized,paraffined and embedding slices were observes under light microscope for pathological changes.RESULTS:Compared w ith t he m odel g roup,nicotiflorin s ignificantly r educed s erum A LT a nd A ST a ctivity o f r ats w ith a cuteliver i njury(P<0.01)a t d oses g reater t han50m g/kg,s ignificantly i mproved l iver h omogenate o f S OD a nd G SH a ctivity a t h igherthan50mg/kg(P<0.05),significantly decreased MDA and NO content in the liver(P<0.05),IL-1,IL-6,reducedinflammation a nd p romoted t he l evel o f I NF-γa nd T NF-αi n s erum o f m ice(P<0.05),reduced l iver a nd s pleen i ndex(P<0.05)at a dose of more than50mg/kg.Liver pathological examinations showed that,compared with the model group,nicotiflorin significantly reduced the degree of liver cell injury.Inflammatory cell infiltration was significantly decreased,including high,medium a nd l ow d ose g roup c ompared t o t he e xtent o f t he d amage a nd r epair.C ONCLUSION:The p rotective e ffect o f D-Gal o nacute l iver i njury i n m ice w as r elated t o t he p romotion o f a ntioxidant e nzymes a nd t he e xpression o f p roinflammatory c ytokines.