苹果褐腐病防治药剂的筛选和残留安全性评价

为筛选苹果褐腐病高效、低毒、低残留农药,采用菌丝生长速率法,测定了8种杀菌剂对苹果褐腐病菌的抑制率。药剂室内抑菌结果表明,戊唑醇、咪鲜胺、代森联、啶酰菌胺、多抗霉素和吡唑醚菌酯对苹果褐腐病菌的抑菌效果较好,菌丝生长抑制率基本在90%以上。基于药剂室内抑菌结果,筛选了6种药剂进行了田间防效试验,结果表明,450 g/L咪鲜胺AS 1000倍液、430 g/L戊唑醇SC 3000倍液、70%代森联WG 300倍液和25%吡唑醚菌酯SC 1000倍液防治效果均在90%以上,50%啶酰菌胺WG 750倍液和10%多抗霉素WP 1000倍液防效较差。农药残留分析结果表明,咪鲜胺、戊唑醇、代森联、吡唑醚菌酯、啶酰菌胺和多抗霉素农药残留量均符合国家食品安全标准。

复方茵丹汤对急性肝内胆汁淤积大鼠肝组织法尼醇受体及多耐药相关蛋白3表达的影响

目的研究复方茵丹汤对α-萘异硫氰酸酯(ANIT)诱导的大鼠急性肝内胆汁淤积的作用机制。方法将108只雄性SD大鼠随机分为正常对照组,模型组,中药复方茵丹汤高、中、低剂量组和阳性药对照组,每组18只,依据取材时间不同又将各组随机分为24、48、72 h 3个亚组,每组6只。模型组、中药组、阳性药对照组大鼠一次性给予2%ANIT按100 mg·kg^(-1)·d^(-1)灌胃造模,正常对照组予等容积色拉油灌胃。造模2 h后,中药高、中、低剂量组分别给予生药含量为24.48 g·kg^(-1)·d^(-1)、12.24 g·kg^(-1)·d^(-1)和6.12 g·kg^(-1)·d^(-1)的复方茵丹汤中药煎剂l ml/100 g体质量灌胃,模型组和正常对照组给予等容积生理盐水灌胃,阳性药对照组给予67.5 mg·kg^(-1)·d^(-1)熊去氧胆酸(UDCA)水溶液l ml/100 g体质量灌胃,均1次/d。造模后分别于24、48、72 h处死相应时相大鼠,采用RT-q-PCR、Western Blot和免疫组化技术检测法尼醇受体(FXR)及多耐药相关蛋白3(MRP3)在大鼠肝组织的表达水平。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果 RT-q-PCR结果显示,与同时相正常组比较,模型组FXR mRNA表达均明显降低(P值均<0.05);与同时相模型组比较,中药各剂量组及UDCA组FXR mRNA表达明显升高(P值均<0.05);中药高剂量组各时相FXR mRNA表达明显高于中、低剂量组(P值均<0.05)。与同时相正常组比较,模型组MRP3 mRNA表达均显著升高(P值均<0.05);与同时相模型组比较,除低剂量组24 h外,中药各剂量组及UDCA组MRP3 mRNA表达均明显升高(P值均<0.05);中药高剂量组48 h、72 h的MRP3 mRNA表达均明显高于中、低剂量组(P值均<0.05)。免疫组化结果显示,与同时相正常组比较,模型组FXR蛋白表达均显著降低(P值均<0.05);与模型组比较,24 h中药各剂量组及各时相UDCA组FXR蛋白表达均显著升高(P值均<0.05);中药高剂量组FXR蛋白表达明显高于同时相中、低剂量组和模型组(P值均<0.05)。与同时相正常组比较,模型组MRP3蛋白表达显著升高(P值均<0.05);与同时相模型组比较,除低剂量24 h外,中药各剂量组及UDCA组MRP3蛋白表达明显升高(P值均<0.05);中药高剂量组MRP3蛋白表达显著高于同时相低剂量组及72 h中剂量组(P值均<0.05)。Western Blot结果显示,与同时相正常组比较,模型组FXR蛋白表达均显著降低(P值均<0.05);与同时相模型组比较,中药各剂量组及UDCA组FXR蛋白表达均显著升高(P值均<0.05);中药高剂量组FXR蛋白表达均显著高于同时相中、低剂量组(P值均<0.05)。与同时相正常组比较,模型组MRP3蛋白表达均显著升高(P值均<0.05);与模型组比较,48 h及72 h中药中、高剂量组及UDCA组MRP3蛋白表达均明显升高(P值均<0.05);中药高剂量组48 h和72 h的MRP3蛋白表达均高于相应的中、低剂量组(P值均<0.05)。结论复方茵丹汤对ANIT灌胃诱导AIC大鼠的治疗作用可能与其上调FXR和MRP3基因、蛋白表达有关。

复方茵丹汤对急性肝内胆汁淤积大鼠肝组织早期生长反应因子-1和细胞间黏附分子-1表达的影响

目的研究复方茵丹汤对α-萘异硫氰酸酯(α-naphthalene isothiocyanate,ANIT)诱导的大鼠急性肝内胆汁淤积作用机制。方法将108只雄性SD大鼠随机分为空白对照组,模型组,中药高、中、低剂量组和阳性药对照组,每组18只,依据取材时间不同又将各组随机分为24、48、72 h 3个亚组,每组6只。模型组、中药组、阳性药对照组大鼠一次性给予2%ANIT按100 mg/(kg·d)灌胃造模,空白对照组予等容积色拉油灌胃。造模2 h后,中药高、中、低剂量组分别给予生药含量为24.48、12.24和6.12 g/(kg·d)的复方茵丹汤中药煎剂1 mL/100 g体重灌胃,模型组和空白对照组给予等容积生理盐水灌胃,阳性药对照组给予67.5 mg/(kg·d)去氧胆酸灌胃,均1次/日。造模后分别于24、48、72 h处死相应时相大鼠,采用RT-q-PCR、Western blot和IHC技术检测早期生长反应因子1(early growth response-1,Egr-1)及细胞间黏附分子-1(intercellular adhesion molecule-1,ICAM-1)在大鼠肝组织的表达水平。结果与同时相空白对照组比较,模型组Egr-1、ICAM-1基因及蛋白表达均明显升高(P<0.01)。与同时相模型组比较,中药各剂量组及阳性药对照组Egr-1及ICAM-1基因及蛋白表达均明显降低(P<0.05,P<0.01)。中药高剂量组Egr-1、ICAM-1基因及蛋白表达明显低于同时相中药中、低剂量组(P<0.05,P<0.01),与阳性药对照组比较差异无统计学意义(P>0.05)。结论复方茵丹汤对ANIT灌胃诱导AIC大鼠的治疗作用可能与其下调Egr-1和ICAM-1基因和蛋白表达有关。

Effect of compound Yindan decoction on alpha-naphthylisothiocyanate-Induced acute intrahepatic cholestasis in rats

OBJECTIVE:To investigate the therapeutic mechanism of compound Yindan decoction (CYD) in a rat model of acute intrahepatic cholestatic (AIC).METHODS:A total of 108 adult male rats were randomly divided into control (n =18) and AIC groups (n =90).AIC was induced in rats using alpha-naphthylisothiocyanate (ANIT)(75 mg/kg,10 mL/kg in corn oil,p.o.).Then,90 AIC rats were randomly divided into five groups:a control group (n =18),a CYD high dose group (n =18),a CYD middle dose group (n =18),a CYD low dose group (n =18),and a ursodeoxycholicacid (UDCA) group (n =18).According to sampling time,each group was subdivided into three subgroups:24 h (n =6),48 h (n =6),and 72 h groups (n =6).The CYD-high,-middle and-low groups were orally administered 24.48,12.24,and 6.12 g.kg-1.d-1 modified CYD,respectively,while the model group was given 20 mL/kg of body weight of distilled water once a day.The UDCA group was given 67.5 mg.kg-1.d-1 UDCA once a day.Radioimmunity assay was used to detect the activity of alanine aminotransferase (ALT),aspartate aminotransferase (AST),alkaline phosphatase (ALP),gamma-glutamyl transpeptidase (GGT) and the levels of total bilirubin (TBil) and indirectbiliruin (DBil) in rats.Reverse transcription quantitative polymerase chain reaction (qRT-PCR),Western blot analysis,and immunohistochemistry were used to detect multidrug resistance-associated protein 2 (MRP2) expression.In vitro,HepG2 hepatocellular carcinoma cells were treated with CYD medicated serum at a concentration of 15 mol/L.MRP2 and retinoid X receptor alpha (RXRα) expression was analyzed by qRT-PCR and Western blotting.RESULTS:Serum levels of ALT,AST,GGT,ALP,TBil,and DBil were significantly reduced in the CYD and positive drug groups compared with the control group (P < 0.05 and P < 0.01,respectively).Pathological changes in rat liver tissues at 72 h in the CYD-high and-medium dose groups and positive drug group were not significant compared with the control group.CYD and UDCA treatment ameliorated ANIT-induced biliary epithelial cell proliferation.Neutrophil infiltration was rare and little focal necrosis was observed in Iobules in the CYD-high and-medium dose groups and UDCA group at 72 h.Compared with the control group,the expression of MRP2 mRNA and MRP2 protein in the liver tissue of the CYD groups was significantly increased (P <0.05 and P < 0.01,respectively).MRP2 expression and RXRα nuclear receptor mRNA and protein levels in the CYD groups were significantly increased compared with the control and UDCA groups (P <0.01).CONCLUSION:CYD can alleviate cholestasis in ANIT-induced AIC rats,and the mechanism underlying this action might involve increases in ALT,AST,GGT,ALP,TBil,and DBil and upregulation of MRP2 and RXRα mRNA and protein levels.