[目的]探讨运脾和络颗粒对糖尿病周围神经病变(diabetic peripheral neuropathy,DPN)大鼠坐骨神经组织发动相关蛋白-1(dynamin-related protein 1,Drp-1)和Caspase-3的干预作用。[方法]链脲佐菌素加高糖高脂造模成功大鼠随机分成模型组...[目的]探讨运脾和络颗粒对糖尿病周围神经病变(diabetic peripheral neuropathy,DPN)大鼠坐骨神经组织发动相关蛋白-1(dynamin-related protein 1,Drp-1)和Caspase-3的干预作用。[方法]链脲佐菌素加高糖高脂造模成功大鼠随机分成模型组、运脾和络颗粒低、中、高剂量组、甲钴胺组,另设正常组;采用葡萄糖氧化酶法测血糖,免疫组化检测各组坐骨神经组织的Drp-1和Caspase-3蛋白的表达。[结果]对大鼠血糖而言,模型组给药前后血糖均高于正常组,差异有统计学意义(P<0.01);与模型组比较,运脾和络颗粒高、中、低剂量组都有一定的降糖作用,差异有统计学意义((P<0.01,P<0.001),其中运脾和络颗粒高剂量组血糖降低最明显。模型组坐骨神经纤维中神经膜、轴突中Drp-1的阳性表达比正常组增加,差异有统计学意义(P<0.01);运脾和络颗粒高、中、低剂量组,甲钴胺组Drp-1的蛋白表达均较模型组有所下降,其中高剂量组减少程度最多,差异有统计学意义(P<0.01)。模型组Caspase-3蛋白表达比正常组增加,差异有统计学意义(P<0.01);运脾和络颗粒高、中、低剂量组,甲钴胺组Caspase-3蛋白表达均较模型组有所下降,其中高剂量组减少程度最多,差异有统计学意义(P<0.01)。运脾和络颗粒各组均能下调DPN模型大鼠坐骨神经组织Drp-1和Caspase-3蛋白表达,运脾和络颗粒高、中剂量组差异有统计学意义(P<0.05,P<0.01),其中以运脾和络颗粒高剂量组最为明显(P<0.01),运脾和络颗粒低剂量组和甲钴胺组虽能下调Drp-1和Caspase-3蛋白表达,但差异无统计学意义(P>0.05)。[结论]运脾和络颗粒可以通过下调DPN大鼠周围神经Drp-1和Caspase-3表达,防治DPN的发生发展,且与剂量有一定的量效关系,这可能是运脾和络颗粒防治DPN的保护作用机制之一。展开更多
This work develops a fluorescence approach for sensitive detection of DNA methyltransferase activity based on endonuclease and rolling circle amplification （RCA） technique. In the presence of DNA adenine methylation...This work develops a fluorescence approach for sensitive detection of DNA methyltransferase activity based on endonuclease and rolling circle amplification （RCA） technique. In the presence of DNA adenine methylation （Dam） MTase, the methylation-responsive sequence of hairpin probe is methylated and cleaved by the methylation-sensitive restriction endonuclease Dpn 1. The products cleaved by restriction endonuclease Dpn I then function as a signal primer to initiate RCA reaction by hybridizing with the circular DNA template. Each RCA product containing thousands of repeated sequences might hybridize with a large number of molecular beacons （detection probes）, resulting in an enhanced fluorescence signal. In the absence of Dam MTase, neither methylation/cleavage nor RCA reaction can be initiated and no fluorescence signal is observed. The proposed method exhibits a dynamic range from 0.5 U/mL to 30 U/mL and a detection limit of 0.18 U/mL. This method can be used for the screening of antimicrobial drugs and has a great potential to be further applied in early clinical diagnosis.展开更多
文摘[目的]探讨运脾和络颗粒对糖尿病周围神经病变(diabetic peripheral neuropathy,DPN)大鼠坐骨神经组织发动相关蛋白-1(dynamin-related protein 1,Drp-1)和Caspase-3的干预作用。[方法]链脲佐菌素加高糖高脂造模成功大鼠随机分成模型组、运脾和络颗粒低、中、高剂量组、甲钴胺组,另设正常组;采用葡萄糖氧化酶法测血糖,免疫组化检测各组坐骨神经组织的Drp-1和Caspase-3蛋白的表达。[结果]对大鼠血糖而言,模型组给药前后血糖均高于正常组,差异有统计学意义(P<0.01);与模型组比较,运脾和络颗粒高、中、低剂量组都有一定的降糖作用,差异有统计学意义((P<0.01,P<0.001),其中运脾和络颗粒高剂量组血糖降低最明显。模型组坐骨神经纤维中神经膜、轴突中Drp-1的阳性表达比正常组增加,差异有统计学意义(P<0.01);运脾和络颗粒高、中、低剂量组,甲钴胺组Drp-1的蛋白表达均较模型组有所下降,其中高剂量组减少程度最多,差异有统计学意义(P<0.01)。模型组Caspase-3蛋白表达比正常组增加,差异有统计学意义(P<0.01);运脾和络颗粒高、中、低剂量组,甲钴胺组Caspase-3蛋白表达均较模型组有所下降,其中高剂量组减少程度最多,差异有统计学意义(P<0.01)。运脾和络颗粒各组均能下调DPN模型大鼠坐骨神经组织Drp-1和Caspase-3蛋白表达,运脾和络颗粒高、中剂量组差异有统计学意义(P<0.05,P<0.01),其中以运脾和络颗粒高剂量组最为明显(P<0.01),运脾和络颗粒低剂量组和甲钴胺组虽能下调Drp-1和Caspase-3蛋白表达,但差异无统计学意义(P>0.05)。[结论]运脾和络颗粒可以通过下调DPN大鼠周围神经Drp-1和Caspase-3表达,防治DPN的发生发展,且与剂量有一定的量效关系,这可能是运脾和络颗粒防治DPN的保护作用机制之一。
基金supported by the National Natural Science Foundation of China(Nos.21190044 and 21175035)National Basic Research Program(No.2011CB911002)International Science&Technology operation Program of China(No.2010DFB30300)
文摘This work develops a fluorescence approach for sensitive detection of DNA methyltransferase activity based on endonuclease and rolling circle amplification （RCA） technique. In the presence of DNA adenine methylation （Dam） MTase, the methylation-responsive sequence of hairpin probe is methylated and cleaved by the methylation-sensitive restriction endonuclease Dpn 1. The products cleaved by restriction endonuclease Dpn I then function as a signal primer to initiate RCA reaction by hybridizing with the circular DNA template. Each RCA product containing thousands of repeated sequences might hybridize with a large number of molecular beacons （detection probes）, resulting in an enhanced fluorescence signal. In the absence of Dam MTase, neither methylation/cleavage nor RCA reaction can be initiated and no fluorescence signal is observed. The proposed method exhibits a dynamic range from 0.5 U/mL to 30 U/mL and a detection limit of 0.18 U/mL. This method can be used for the screening of antimicrobial drugs and has a great potential to be further applied in early clinical diagnosis.
