肝疾病,酒精性%核因子κB%茶多酚
肝疾病,酒精性%覈因子κB%茶多酚
간질병,주정성%핵인자κB%다다분
Liver diseases,alcoholic%Nuclear factor kappa B%Tea polyphenols
目的 观察茶多酚对酒精性肝病肝脏核因子-κB(NF-κB)活性调节作用.方法 将22只Wistar大鼠随机分为3组:对照组6只,普通饲料喂养;模型组7只,每天给予大鼠体积分数56%白酒8 ml/kg灌胃,每周递增2ml/kg直至总剂量达每天16ml/kg造模;茶多酚组9只,在模型组的基础上给予茶多酚0.25 g/kg加入白酒中灌胃,7周后处死大鼠.同时培养人正常肝细胞株L02,分为5组:对照组常规培养;模型组:加入0.6%乙醇;预防组:先加入200μg/ml茶多酚培养3 d后再加入0.6%乙醇培养;干预组:同时加入0.6%乙醇和200 μg/ml茶多酚培养;治疗组:先加入0.6%乙醇培养3 d后再加入200μg/ml茶多酚培养.各组细胞均以7d为周期传代,处理4周.实验重复3次.光镜观察肝细胞改变,分光光度计测各组大鼠血清丙二醛及L02细胞活性氧含量,RT-PCR测各组NF-κB和NF-κB抑制因子(I κB)表达;凝胶迁移电泳测各组NF-κB活性变化.结果 NF-κB mRNA表达:对照组为0.53±0.01、模型组为1.15±0.03、茶多酚组为0.58±0.16,模型组明显高于对照组和茶多酚组,F=2431.117,P<0.01,差异有统计学意义.大鼠肝组织NF-κB活性:模型组DNA染色:1410.78±22.19,蛋白染色:1426.08±33.15,对照组DNA染色:419.84±8.32,蛋白染色:419.99±7.06,模型组NF-κB活性比对照组强,P<0.01,差异有统计学意义.茶多酚DNA染色:669.85±41.34,蛋白染色:675.35±18.27,较模型组减弱,P<0.01,差异有统计学意义.L02细胞NF-κB mRNA表达,对照组、模型组、预防组、干预组、治疗组分别0.56±0.01、0.76±0.03,0.60±0.03、0.59±0.01、0.59±0.01,预防组、干预组、治疗组比模型组降低,F=46.353,P<0.01,差异有统计学意义.L02细胞NF-κB活性对照组、模型组、预防组,干预组、治疗组DNA染色分别为:344.42±11.37、849.94±12.45、713.07±11.91,710.79±14.99和693.45±71.69;蛋白染色分别为371.20±13.51、925.96±5.78、758.88±34.65,753.07±76.78和725.77±36.09,预防组、干预组、治疗组比模型组减弱,P<0.01,差异有统计学意义.结论 茶多酚对预防、干预和治疗酒精性肝病均有一定的作用,其作用可能是通过抑制NF-κB的活化实现.
目的 觀察茶多酚對酒精性肝病肝髒覈因子-κB(NF-κB)活性調節作用.方法 將22隻Wistar大鼠隨機分為3組:對照組6隻,普通飼料餵養;模型組7隻,每天給予大鼠體積分數56%白酒8 ml/kg灌胃,每週遞增2ml/kg直至總劑量達每天16ml/kg造模;茶多酚組9隻,在模型組的基礎上給予茶多酚0.25 g/kg加入白酒中灌胃,7週後處死大鼠.同時培養人正常肝細胞株L02,分為5組:對照組常規培養;模型組:加入0.6%乙醇;預防組:先加入200μg/ml茶多酚培養3 d後再加入0.6%乙醇培養;榦預組:同時加入0.6%乙醇和200 μg/ml茶多酚培養;治療組:先加入0.6%乙醇培養3 d後再加入200μg/ml茶多酚培養.各組細胞均以7d為週期傳代,處理4週.實驗重複3次.光鏡觀察肝細胞改變,分光光度計測各組大鼠血清丙二醛及L02細胞活性氧含量,RT-PCR測各組NF-κB和NF-κB抑製因子(I κB)錶達;凝膠遷移電泳測各組NF-κB活性變化.結果 NF-κB mRNA錶達:對照組為0.53±0.01、模型組為1.15±0.03、茶多酚組為0.58±0.16,模型組明顯高于對照組和茶多酚組,F=2431.117,P<0.01,差異有統計學意義.大鼠肝組織NF-κB活性:模型組DNA染色:1410.78±22.19,蛋白染色:1426.08±33.15,對照組DNA染色:419.84±8.32,蛋白染色:419.99±7.06,模型組NF-κB活性比對照組彊,P<0.01,差異有統計學意義.茶多酚DNA染色:669.85±41.34,蛋白染色:675.35±18.27,較模型組減弱,P<0.01,差異有統計學意義.L02細胞NF-κB mRNA錶達,對照組、模型組、預防組、榦預組、治療組分彆0.56±0.01、0.76±0.03,0.60±0.03、0.59±0.01、0.59±0.01,預防組、榦預組、治療組比模型組降低,F=46.353,P<0.01,差異有統計學意義.L02細胞NF-κB活性對照組、模型組、預防組,榦預組、治療組DNA染色分彆為:344.42±11.37、849.94±12.45、713.07±11.91,710.79±14.99和693.45±71.69;蛋白染色分彆為371.20±13.51、925.96±5.78、758.88±34.65,753.07±76.78和725.77±36.09,預防組、榦預組、治療組比模型組減弱,P<0.01,差異有統計學意義.結論 茶多酚對預防、榦預和治療酒精性肝病均有一定的作用,其作用可能是通過抑製NF-κB的活化實現.
목적 관찰다다분대주정성간병간장핵인자-κB(NF-κB)활성조절작용.방법 장22지Wistar대서수궤분위3조:대조조6지,보통사료위양;모형조7지,매천급여대서체적분수56%백주8 ml/kg관위,매주체증2ml/kg직지총제량체매천16ml/kg조모;다다분조9지,재모형조적기출상급여다다분0.25 g/kg가입백주중관위,7주후처사대서.동시배양인정상간세포주L02,분위5조:대조조상규배양;모형조:가입0.6%을순;예방조:선가입200μg/ml다다분배양3 d후재가입0.6%을순배양;간예조:동시가입0.6%을순화200 μg/ml다다분배양;치료조:선가입0.6%을순배양3 d후재가입200μg/ml다다분배양.각조세포균이7d위주기전대,처리4주.실험중복3차.광경관찰간세포개변,분광광도계측각조대서혈청병이철급L02세포활성양함량,RT-PCR측각조NF-κB화NF-κB억제인자(I κB)표체;응효천이전영측각조NF-κB활성변화.결과 NF-κB mRNA표체:대조조위0.53±0.01、모형조위1.15±0.03、다다분조위0.58±0.16,모형조명현고우대조조화다다분조,F=2431.117,P<0.01,차이유통계학의의.대서간조직NF-κB활성:모형조DNA염색:1410.78±22.19,단백염색:1426.08±33.15,대조조DNA염색:419.84±8.32,단백염색:419.99±7.06,모형조NF-κB활성비대조조강,P<0.01,차이유통계학의의.다다분DNA염색:669.85±41.34,단백염색:675.35±18.27,교모형조감약,P<0.01,차이유통계학의의.L02세포NF-κB mRNA표체,대조조、모형조、예방조、간예조、치료조분별0.56±0.01、0.76±0.03,0.60±0.03、0.59±0.01、0.59±0.01,예방조、간예조、치료조비모형조강저,F=46.353,P<0.01,차이유통계학의의.L02세포NF-κB활성대조조、모형조、예방조,간예조、치료조DNA염색분별위:344.42±11.37、849.94±12.45、713.07±11.91,710.79±14.99화693.45±71.69;단백염색분별위371.20±13.51、925.96±5.78、758.88±34.65,753.07±76.78화725.77±36.09,예방조、간예조、치료조비모형조감약,P<0.01,차이유통계학의의.결론 다다분대예방、간예화치료주정성간병균유일정적작용,기작용가능시통과억제NF-κB적활화실현.
Objective To investigate the effect of tea polyphenols (TP) on expression of nuclear factor kappa B (NF- κB) in rats with alcoholic liver diseases and in cells treated with alcohol. Methods 22 female Wistar rats were randomly divided into three groups: a control group, an alcohol model group and a TP plus alcohol group. All treatments were injected into stomach through intragastric tube. L02 cells were divided into five groups: a control group, an alcohol treated group, a prevention group (cells were treated with TP for 3 days, and then treated with alcohol), an intervention group (cells treated with TP and alcohol), and a therapeutic group (cells were treated with alcohol for 3 days, and then treated with TP). Histopathology was observed under light microscope (LM); serum MDA, ROS in cells were quantified by optical density measurement ; the expression of NF- κB and I k B was determined by RT-PCR; and the activity of NF- κB was checked with Electrophoretic Mobility Shift Assay (EMSA). Results LM indicated hepatocytes were injured obviously in the model group. Serum MDA and cells ROS in TP treated groups were significantly lower than the alcohol treated group. The level of NF- κB mRNA expression in TP treated groups(rats: 0.58 ±0.16, cells: 0.60 ± 0.03, 0.59 ± 0.01,0.59 ± 0.01) were significantly lower than the alcohol treated group (rats: 1.15 ± 0.03, cells: 0.76 ± 0.03) (P < 0.01), the level of I κB mRNA expression in the prevention group, intervention group, and therapeutic group (0.51 ± 0.01, 0.50 + 0.01, 0.50 ± 0.12) were significantly higher than the alcohol treated group (0.61 ± 0.03) (P < 0.05), the difference among the three groups was not significant (P > 0.05). The activity of NF- κB in TP treated rats(DNA stain: 669.85 + 41.34, Protein stain: 675.35 ± 18.27) was significantly lower than the alcohol treated rats(DNA stain: 1410.78 ± 22.19,Protein stain:1426.08 ± 33.15) (P < 0.01); NF- κB activity in cells of the prevention, intervention, therapeutic groups (DNA stain: 713.07 ± 11.91, 710.79 ± 14.99, 693.45 ± 71.69; Protein stain: 758.88 ±34.65, 753.07 ± 76.78, 725.77 + 36.09) was significantly lower than the alcohol treated cells (DNA stain:849.94 ± 12.45, Protein stain: 925.96 ± 5.78) (P < 0.01), the difference among the three TP treated groups was not significant (P > 0.01). Conclusion TP can alleviate and prevent alcohol-induced liver injury via inhibiting NF- κB activation.
