Objective: Observing the expression changes of serum proteome in model rats after intervention of the Granules of Eliminating Phlegm and Removing Blood Stasis (豁痰祛瘀颗粒 also known as GEPRB), screening out and iden...Objective: Observing the expression changes of serum proteome in model rats after intervention of the Granules of Eliminating Phlegm and Removing Blood Stasis (豁痰祛瘀颗粒 also known as GEPRB), screening out and identifying the differentially expressed proteins by mass spectrometry and bioinformatics analysis, discussing the molecular mechanism of control the Diabetes deafness by GEPRB. Methods: By use of proteomics technology, the serum protein serum proteome of the control group, model control group, Duxil and each observation group were observed for 2-DE gel pattern matching, and the difference in the relative content of 2 times was chosen for the differentially expressed proteins. Identification of differentially expressed proteins by MALDI-TOF MS/MS, the authors further analysis the phosphorylation, subcellular localization, interaction, direct regulation, and transmembrane of the differences proteins by the way of bioinformatics analysis. Sixty SPF level SD rats elected in diabetic rats model group (abbreviated as DM group) were be randomly divided into 5 groups based on random number sheet, namely model control group, positive drug control group (Du-ke-xi group) and Mai-tong-fang high, medium and low dose group respectively. In addition, set of normal control group. 10 rats in each group. Results: By Coomassie brilliant blue staining, identified 51 differential protein spots dug from 2-D gel by mass spectrometry, successfully identified 13 non-redundant proteins. Most of the identified proteins were secreted protein and belong to different protein families. There were about 12 proteins have the transmembrane region from the authors’ result, ten of them were plasma membrane proteins. Conclusion: It’s suggesting that 13 differential proteins is most likely the protein response to GEPRB in vivo, these proteins may play key role for the treatment of GEPRB to Diabetes deafness. The two highly differentially expressed proteins Apolipoprotein E (apoE) and C3 may be a potential drug target of GEPRB.展开更多
目的:探讨痰瘀共治方对大鼠动脉粥样硬化(atherosclerosis,AS)斑块的影响及其可能机制。方法:12周龄雄性Wistar大鼠85只,随机分为5组:①正常组(the control group,C,n=20),②模型组(the model group,M,n=20),③罗格列酮组(the Rosiglita...目的:探讨痰瘀共治方对大鼠动脉粥样硬化(atherosclerosis,AS)斑块的影响及其可能机制。方法:12周龄雄性Wistar大鼠85只,随机分为5组:①正常组(the control group,C,n=20),②模型组(the model group,M,n=20),③罗格列酮组(the Rosiglitazone group,RSG,n=15),④痰瘀共治低剂量组(TYD,n=15),⑤痰瘀共治高剂量组(TYG,n=15)。采用高脂饲料喂饲法加维生素D3负荷法复制大鼠AS模型,6周后取C组及M组大鼠各5只,以主动脉HE染色切片发现AS斑块作为造模成功指标。造模成功后除正常组喂饲基础饲料,余组继续喂饲高脂饲料,并分别加予生理盐水[10mL/(kg·d)]、罗格列酮(3mg/kg)、痰瘀共治方流浸膏高剂量(0.8g/mL)、痰瘀共治方流浸膏低剂量(1.6g/mL)灌胃治疗6周后处死,HE染色观察大鼠主动脉形态学变化,免疫组织化学染色法观察主动脉过氧化物酶体增值物激活受体γ(PPARγ)蛋白表达的变化,并检测空腹血糖(fasting plasma glu-cose,FPG)、空腹胰岛素(fasting insulin,FINS),计算胰岛素抵抗指数(HOMA-IR)。结果:高脂饮食加维生素D3负荷6周后造模成功,光镜下观察可见主动脉典型的粥样硬化斑块;灌胃治疗6周后,与M组比,各治疗组大鼠FPG、FISN、HOMA-IR水平均降低(P<0.05或P<0.01),HE染色见斑块明显消退,PPARγ蛋白表达平均光密度值明显减弱,以TYG组(1.08±0.03)作用显著(P=0.0025),低于TYD组(2.84±0.07)(P=0.034),与RSG组(1.12±0.05)比差异亦没有统计学意义(P=0.427),同时各治疗组与C组比差异没有统计学意义(P>0.05)。结论:痰瘀共治方可能通过激活PPARγ,改善胰岛素抵抗起到抗动脉粥样硬化的作用。展开更多
文摘Objective: Observing the expression changes of serum proteome in model rats after intervention of the Granules of Eliminating Phlegm and Removing Blood Stasis (豁痰祛瘀颗粒 also known as GEPRB), screening out and identifying the differentially expressed proteins by mass spectrometry and bioinformatics analysis, discussing the molecular mechanism of control the Diabetes deafness by GEPRB. Methods: By use of proteomics technology, the serum protein serum proteome of the control group, model control group, Duxil and each observation group were observed for 2-DE gel pattern matching, and the difference in the relative content of 2 times was chosen for the differentially expressed proteins. Identification of differentially expressed proteins by MALDI-TOF MS/MS, the authors further analysis the phosphorylation, subcellular localization, interaction, direct regulation, and transmembrane of the differences proteins by the way of bioinformatics analysis. Sixty SPF level SD rats elected in diabetic rats model group (abbreviated as DM group) were be randomly divided into 5 groups based on random number sheet, namely model control group, positive drug control group (Du-ke-xi group) and Mai-tong-fang high, medium and low dose group respectively. In addition, set of normal control group. 10 rats in each group. Results: By Coomassie brilliant blue staining, identified 51 differential protein spots dug from 2-D gel by mass spectrometry, successfully identified 13 non-redundant proteins. Most of the identified proteins were secreted protein and belong to different protein families. There were about 12 proteins have the transmembrane region from the authors’ result, ten of them were plasma membrane proteins. Conclusion: It’s suggesting that 13 differential proteins is most likely the protein response to GEPRB in vivo, these proteins may play key role for the treatment of GEPRB to Diabetes deafness. The two highly differentially expressed proteins Apolipoprotein E (apoE) and C3 may be a potential drug target of GEPRB.
文摘目的:探讨痰瘀共治方对大鼠动脉粥样硬化(atherosclerosis,AS)斑块的影响及其可能机制。方法:12周龄雄性Wistar大鼠85只,随机分为5组:①正常组(the control group,C,n=20),②模型组(the model group,M,n=20),③罗格列酮组(the Rosiglitazone group,RSG,n=15),④痰瘀共治低剂量组(TYD,n=15),⑤痰瘀共治高剂量组(TYG,n=15)。采用高脂饲料喂饲法加维生素D3负荷法复制大鼠AS模型,6周后取C组及M组大鼠各5只,以主动脉HE染色切片发现AS斑块作为造模成功指标。造模成功后除正常组喂饲基础饲料,余组继续喂饲高脂饲料,并分别加予生理盐水[10mL/(kg·d)]、罗格列酮(3mg/kg)、痰瘀共治方流浸膏高剂量(0.8g/mL)、痰瘀共治方流浸膏低剂量(1.6g/mL)灌胃治疗6周后处死,HE染色观察大鼠主动脉形态学变化,免疫组织化学染色法观察主动脉过氧化物酶体增值物激活受体γ(PPARγ)蛋白表达的变化,并检测空腹血糖(fasting plasma glu-cose,FPG)、空腹胰岛素(fasting insulin,FINS),计算胰岛素抵抗指数(HOMA-IR)。结果:高脂饮食加维生素D3负荷6周后造模成功,光镜下观察可见主动脉典型的粥样硬化斑块;灌胃治疗6周后,与M组比,各治疗组大鼠FPG、FISN、HOMA-IR水平均降低(P<0.05或P<0.01),HE染色见斑块明显消退,PPARγ蛋白表达平均光密度值明显减弱,以TYG组(1.08±0.03)作用显著(P=0.0025),低于TYD组(2.84±0.07)(P=0.034),与RSG组(1.12±0.05)比差异亦没有统计学意义(P=0.427),同时各治疗组与C组比差异没有统计学意义(P>0.05)。结论:痰瘀共治方可能通过激活PPARγ,改善胰岛素抵抗起到抗动脉粥样硬化的作用。
