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1、.中文摘要目的：观察研究加味四逆散及其拆方药物血清对肝星状细胞HSC-T6JAK2-STAT3信号通路的影响,探讨加味四逆散及拆方抗肝纤维化作用机理,分析研究不同药组影响肝纤维化的靶位,为选择针对不同靶位最有治疗优势的药物,重新组合成更优化的抗肝纤维化中药复方提供依据。方法：加味四逆散全方及其拆方药物煎剂给大鼠灌胃 7d；制备各含药血清；培养肝星状细胞；用瘦素刺激 HSC-T6 使其增殖；蛋白印迹试验检测各药物血清对增殖后的肝星状细胞100ng/ml 的瘦素刺激 HSC-T6JAK2-STAT3 蛋白表达的影响效果。结果：1.加味四逆散全方药物血清对增殖后的肝星状细胞下同作用6、12、24h后
2、JAK2表达的影响结果显示,加味四逆散组的蛋白条带颜色较正常血清对照组浅,JAK2表达明显降低；6h与 12h、24h三个时段JAK2表达的蛋白条带颜色无明显差异,显示加味四逆散药物血清能明显降低JAK2蛋白表达水平图1图2；2.加味四逆散全方药物血清对肝星状细胞作用6、12、24h后STAT3表达的影响结果显示,加味四逆散组药物血清作用6h后,蛋白条带颜色开始变浅,STAT3水平逐渐降低,至24hSTAT3蛋白表达水平降到最低,与正常对照组比较降低明显,显示加味四逆散药物血清能明显降低STAT3蛋白表达水平图3；3.加味四逆散组、秋水仙碱组、正常血清组各药物血清对肝星状细胞作用12h后,其S
3、TAT3蛋白表达的蛋白条带颜色与正常血清对照组比较明显变浅,有显著性差异：而加味四逆散组与秋水仙碱组蛋白条带颜色相比未见明显差异,显示加味四逆散药物血清能明显降低STAT3蛋白表达水平图4；4.疏肝药物组、活血药物组、健脾药物组各药物血清对肝星状细胞作用12h后,JAK2,STAT3表达的结果显示,拆方药物血清三组与正常血清对照组蛋白条带颜色相比,均明显变浅,有显著性差异；拆方药物血清三组与秋水仙碱组蛋白条带颜色比较未见明显差异；拆方三组与加味四逆散全方组相比蛋白条带颜色稍深,显示其降低JAK2,STAT3表达的效果不如加味四逆散全方；拆方三组之间蛋白条带颜色略存在着差异,疏肝组浅于活血组浅于
4、健脾组,显示拆方三组药物血清均能降低JAK2,STAT3表达的水平,但疏肝组优于活血组优于健脾组图5。结论：1.加味四逆散全方及其拆方各组药物均有降低增殖后的肝星状细胞瘦41 / 41.素刺激HSC-T6JAK2、STAT3蛋白的表达,抑制JAK2-STAT3信号通路信号转导的作用,说明加味四逆散全方及其拆方各组药物抗肝纤维化的主要作用之一可能是通过降低JAK2、STAT3蛋白表达,阻断JAK2-STAT3经典通路的信号转导来实现的。2.加味四逆散全方降低JAK2、STAT3蛋白表达,阻断JAK2-STAT3经典通路信号转导的作用显著优于拆方中单一方的效果,说明综合运用疏肝、健脾、活血的方药治
5、疗肝纤维化可能会取得比单一疏肝或健脾或活血方药更好的疗效。关键词：加味四逆散；拆方；瘦素；肝星状细胞；JAK2-STAT3信号通路.JiaWei Sini Decoction and its ingredients on hepatic stellateCell JAK2-STAT3 signaling pathways affect in signaltransductionAbstractObjective: To study Jiawei Sini Decoction and its separate components of drugserum on hepatic stellate
6、 cells JAK2-STAT3 signaling pathway, ofJiawei Sini Decoction and its separate mechanism of liver fibrosis, analysisof different Medicines affecting liver fibrosis in target site, for differenttarget site for the selection of the most dominant drug treatment, re-combinedto more optimized liver fibros
7、is provide the basis for traditional Chinesemedicine compound.Methods: Modified all Sini decoction Recipe drugs administered to the rats7d; prepare the drug-containing serum; hepatic stellate cell; with leptin tostimulate the proliferation of HSC-T6; Western blot test to detect After thedrug serum o
8、n the proliferation of hepatic stellate cells JAK2-STAT3 protein expression results.Results: 1.Jiawei Sini Decoction all parties after the serum on theproliferation of hepatic stellate cells the role of 6,12,24 h afterthe JAK2 expression results, Jiawei Sini Decoction group protein band colorthan no
9、rmal serum control light, JAK2 expression was reduced; 6h and 12h, 24hthe expression of the three periods JAK2 protein band was no significantdifference in color, showing Jiawei Sini Decoction significantly lower serumprotein levels of JAK2 ;2.Jiawei Sini Decoction all-serum on hepatic stellate cell
10、s 6,12,24 hafter the expression of STAT3 results, Jiawei Sini Decoction group after 6hthe role of drugs in serum, protein bands began to lighter colors, STAT3 leveldecreased protein levels to minimize 24hSTAT3, compared with the control group.decreasedsignificantly,indicatingJiaweiSiniDecoctioncould
11、.significantly reduce the serum levels of STAT3 protein expression ;3.Jiawei Sini Decoction group, colchicine group, the normal serum groupserum on hepatic stellate cells after 12h, the STAT3 protein in protein bandsof color and normal serum control group significantly lighter, there wassignificant
12、difference: while Jiawei Sini Decoction group and colchicinegroup protein band was no significant difference compared with the color,display Jiawei Sini Decoction could significantly reduce the serum levels ofSTAT3 protein expression ;4.Traditional medicine group, activating the drug group, spleen a
13、ndserum of drug group on the role of hepatic stellate cells after 12h, JAK2,STAT3 expression showed that the three separate components serum and normalcontrol group, serum protein band color compared were significantly lighter,there was significant difference; separate components serum colchicine gr
14、oupof three protein bands with no obvious difference in color; separatecomponents and Jiawei Sini Decoction three all-a protein compared with aslightly darker color, indicating the decrease JAK2, STAT3 expression aseffective as the whole side of Jiawei Sini Decoction; split side protein bandsbetween
15、 the three groups there is a little different color, light on the bloodgroup Traditional group light in the spleen group, shows three separatecomponents could decrease serum JAK2, STAT3 expression levels, but better thanthe Traditional group was better than blood group spleen group .Conclusion: 1.Jiawei Sini Decoction All Recipe in each group after the drugshave reduced the proliferation of hepatic stellate cells JAK2, STAT3 protein expression, inhibition
