Depression is a psychological disease with no particularly effective therapy currently available.In the present study,we aimed to examine the antidepressive activity of a pharmaceutical Chinese medicine called TaiZi(T...Depression is a psychological disease with no particularly effective therapy currently available.In the present study,we aimed to examine the antidepressive activity of a pharmaceutical Chinese medicine called TaiZi(TZ)capsule,consisting of total polysaccharides of Radix Pseudostellariae and total flavonoids of both Radix Pueraria and Herba Epimedii.A tail suspension test and forced swimming test were performed to assess the effect of TZ in vivo.A plasmid of TPH2(tryptophan hydroxylase-2)was constructed to determine the exact target of TZ in vitro.In addition,mRNA expression was detected using a real-time PCR assay,and the protein expression was investigated using a Western blotting analysis.The results showed that TZ had an anti-depression effect in mouse and rat models with increased serotonin in the brain,and in the upregulation of mRNA and protein expression of TPH2 in the brain simultaneously by inhibition of NRSF(neuron restrictive silencer factor)expression because NRSF could bind to NRSE(neuron restrictive silencer element)to repress TPH2 transcription during the depression conditions.Icariin could bind to NRSE directly and block NRSF protein toward to NRSE for TPH2 inhibition.Therefore,we concluded that TZ had potential antidepressive effects because it could ameliorat the depression-like behavior in the animals,and the underlying mechanism of the effect was involved in NRSF/NRSE-TPH2 signaling.Icariin was identified as the active component of TZ.This study provided a new perspective for the development of antidepression drugs(Chinese medicines)based on NRSF/NRSE-TPH2 signaling.展开更多
Escherichia coli(E.coli)is often used to produce recombinant proteins rapidly with high yield.However,the bacteria expresses non-target proteins unexpectedly in many cases,some of which may later be proven useful.Full...Escherichia coli(E.coli)is often used to produce recombinant proteins rapidly with high yield.However,the bacteria expresses non-target proteins unexpectedly in many cases,some of which may later be proven useful.Full characterization of these unpredicted proteins are usually expensive and time-consuming.In this study,we used E.coli to express neuron-restrictive silencer factor/REl-silencing transcription factor(NRSF/REST)functional motif ZnF2-8,which is involved in the interaction of NRSF/REST with neuron-restrictive silencer element(NRSE/RE1)dsDNA or small non-coding dsRNA for neuron gene transcriptional repression or activation.Overexpression of a non-target protein was observed.Two-dimensional 1H-15N HSQC NMR spectroscopy,X-ray crystallography and other biochemical assays were used in combination to characterize the non-target protein to beβ-lactamase.展开更多
基金The Student Research Training(SRT)Program of Tsinghua University(Grant No.1522T0221)the National Natural Science Foundation of China(Grant No.81374006 and 81073092)。
文摘Depression is a psychological disease with no particularly effective therapy currently available.In the present study,we aimed to examine the antidepressive activity of a pharmaceutical Chinese medicine called TaiZi(TZ)capsule,consisting of total polysaccharides of Radix Pseudostellariae and total flavonoids of both Radix Pueraria and Herba Epimedii.A tail suspension test and forced swimming test were performed to assess the effect of TZ in vivo.A plasmid of TPH2(tryptophan hydroxylase-2)was constructed to determine the exact target of TZ in vitro.In addition,mRNA expression was detected using a real-time PCR assay,and the protein expression was investigated using a Western blotting analysis.The results showed that TZ had an anti-depression effect in mouse and rat models with increased serotonin in the brain,and in the upregulation of mRNA and protein expression of TPH2 in the brain simultaneously by inhibition of NRSF(neuron restrictive silencer factor)expression because NRSF could bind to NRSE(neuron restrictive silencer element)to repress TPH2 transcription during the depression conditions.Icariin could bind to NRSE directly and block NRSF protein toward to NRSE for TPH2 inhibition.Therefore,we concluded that TZ had potential antidepressive effects because it could ameliorat the depression-like behavior in the animals,and the underlying mechanism of the effect was involved in NRSF/NRSE-TPH2 signaling.Icariin was identified as the active component of TZ.This study provided a new perspective for the development of antidepression drugs(Chinese medicines)based on NRSF/NRSE-TPH2 signaling.
基金The National Basic Research Program of China(2011CB966300)the National Natural Science Foundation of China(30970595,21472229 and 20921091)
文摘Escherichia coli(E.coli)is often used to produce recombinant proteins rapidly with high yield.However,the bacteria expresses non-target proteins unexpectedly in many cases,some of which may later be proven useful.Full characterization of these unpredicted proteins are usually expensive and time-consuming.In this study,we used E.coli to express neuron-restrictive silencer factor/REl-silencing transcription factor(NRSF/REST)functional motif ZnF2-8,which is involved in the interaction of NRSF/REST with neuron-restrictive silencer element(NRSE/RE1)dsDNA or small non-coding dsRNA for neuron gene transcriptional repression or activation.Overexpression of a non-target protein was observed.Two-dimensional 1H-15N HSQC NMR spectroscopy,X-ray crystallography and other biochemical assays were used in combination to characterize the non-target protein to beβ-lactamase.
