Background Histone deacetylase inhibitors （HDACIs） have been reported to induce apoptosis in cancer cells. The effects of trichostatin A （TSA） on gastric cancer cells have not been well characterized. This study w...Background Histone deacetylase inhibitors （HDACIs） have been reported to induce apoptosis in cancer cells. The effects of trichostatin A （TSA） on gastric cancer cells have not been well characterized. This study was aimed to explore the effects and mechanisms of TSA on human gastric cancer SGC-7901 cells. Methods The cells were treated with TSA and analyzed by cell proliferation assay, Western blot, TUNEL assay, flow cytometry by fluorescein isothiocyanate （FITC） conjugated with Annexin V and PI staining, immunofluorescence analysis, analysis of subcellular fractionation, gene chips and real time polymerase chain reaction （PCR）. Results TSA could inhibit cell growth and induced apoptosis in gastric cancer SGC-7901 cells through the regulation of apoptosis-related genes, such as Bcl-2, Bax and survivin. Further study indicated that the pan-caspase inhibitor z-VAD-fmk did not inhibit the apoptosis induced by TSA, and we did not observe the cleavage of poly ADP ribose polymerase （PARP） after TSA treatment too. In addition, apoptosis inducing factor （AIF） and EndoG were found to translocate from mitochondria to nucleus in the immunofluorescence assay and the Western analysis of subcellular fractionation confirmed the result of immunofluorescence assay. Conclusions The apoptosis induced by TSA in gastric cancer SGC-7901 cells involves a caspase-independent pathway.展开更多
Parkinson’s disease(PD)is a multifactorial disorder of the nervous system where a progressive loss of dopaminergic neurons exist.However,the pathogenesis of PD remains undefined,which becomes the main limitation for ...Parkinson’s disease(PD)is a multifactorial disorder of the nervous system where a progressive loss of dopaminergic neurons exist.However,the pathogenesis of PD remains undefined,which becomes the main limitation for the development of clinical PD treatment.Demethylenetetrahydroberberine(DMTHB)is a novel derivative of natural product berberine.This study was aimed to explore the neuroprotective effects and pharmacological mechanism of DMTHB on Parkinson’s disease using C57BL/6 mice.A PD model of mice was induced by administration of MPTP(20 mg·kg^(-1))and probenecid(200 mg·kg^(-1))twice per week for five weeks.The mice were administered with DMTHB daily by gavage at the dose of 5 and 50 mg·kg^(-1)for one-week prophylactic treatment and five-week theraputic treatment.The therapeutic effects of DMTHB were evaluated by behavior tests(the open field,rotarod and pole tests),immunohistochemical staining of tyrosine hydroxylase(TH),Nissl staining and biochemical assays.The molecular mechanisms of DMTHB on the key biomarkers of PD pathological states were analyzed by Western blot(WB)and qRT-PCR.DMTHB treatment alleviated the behavioral disorder induced by MPTP-probenecid.Nissl staining and TH staining showed that the damage of dopaminergic neurons in the substantia nigra was remarkably suppressed by DMTHB treatment.Western blot results showed that the ratio of Bcl-2/Bax and TH increased,but the level ofα-synuclein(α-syn)was remarkably reduced,which indicated that the apoptosis of dopaminergic neurons in mice was significantly reduced.The protein phosphorylation of p-PI3K,p-AKT and p-mTOR also increased about 2-fold,compared with the model group.Furthermore,qRT-PCR results demonstrated that the mRNA levels of pro-inflammatory cytokines,IL-1βand TNF-α,were reduced,but the level of anti-inflammatory cytokine IL-10 increased after DMTHB treatment.Finally,the cellular assay displayed that DMTHB was also a strong antioxidant to protect neuron cell line PC12 by scavenging ROS.In this study,we demonstrated DMTHB alleviates the behavioral disorder and protects dopaminergic neurons through multiple-target effects includubg anti-apoptotic,anti-inflammatory and antioxidant effects.展开更多
文摘Background Histone deacetylase inhibitors （HDACIs） have been reported to induce apoptosis in cancer cells. The effects of trichostatin A （TSA） on gastric cancer cells have not been well characterized. This study was aimed to explore the effects and mechanisms of TSA on human gastric cancer SGC-7901 cells. Methods The cells were treated with TSA and analyzed by cell proliferation assay, Western blot, TUNEL assay, flow cytometry by fluorescein isothiocyanate （FITC） conjugated with Annexin V and PI staining, immunofluorescence analysis, analysis of subcellular fractionation, gene chips and real time polymerase chain reaction （PCR）. Results TSA could inhibit cell growth and induced apoptosis in gastric cancer SGC-7901 cells through the regulation of apoptosis-related genes, such as Bcl-2, Bax and survivin. Further study indicated that the pan-caspase inhibitor z-VAD-fmk did not inhibit the apoptosis induced by TSA, and we did not observe the cleavage of poly ADP ribose polymerase （PARP） after TSA treatment too. In addition, apoptosis inducing factor （AIF） and EndoG were found to translocate from mitochondria to nucleus in the immunofluorescence assay and the Western analysis of subcellular fractionation confirmed the result of immunofluorescence assay. Conclusions The apoptosis induced by TSA in gastric cancer SGC-7901 cells involves a caspase-independent pathway.
基金supported by the National Natural Science Foundation of China(No.81573484)the Postgraduate Research&Practice Innovation Program of Jiangsu Province(KYCX17_0705)+1 种基金the Opening Project of Shanghai Key Laboratory of Complex Prescription(Shanghai University of Traditional Chinese Medicine)(14DZ2271000)a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)。
文摘Parkinson’s disease(PD)is a multifactorial disorder of the nervous system where a progressive loss of dopaminergic neurons exist.However,the pathogenesis of PD remains undefined,which becomes the main limitation for the development of clinical PD treatment.Demethylenetetrahydroberberine(DMTHB)is a novel derivative of natural product berberine.This study was aimed to explore the neuroprotective effects and pharmacological mechanism of DMTHB on Parkinson’s disease using C57BL/6 mice.A PD model of mice was induced by administration of MPTP(20 mg·kg^(-1))and probenecid(200 mg·kg^(-1))twice per week for five weeks.The mice were administered with DMTHB daily by gavage at the dose of 5 and 50 mg·kg^(-1)for one-week prophylactic treatment and five-week theraputic treatment.The therapeutic effects of DMTHB were evaluated by behavior tests(the open field,rotarod and pole tests),immunohistochemical staining of tyrosine hydroxylase(TH),Nissl staining and biochemical assays.The molecular mechanisms of DMTHB on the key biomarkers of PD pathological states were analyzed by Western blot(WB)and qRT-PCR.DMTHB treatment alleviated the behavioral disorder induced by MPTP-probenecid.Nissl staining and TH staining showed that the damage of dopaminergic neurons in the substantia nigra was remarkably suppressed by DMTHB treatment.Western blot results showed that the ratio of Bcl-2/Bax and TH increased,but the level ofα-synuclein(α-syn)was remarkably reduced,which indicated that the apoptosis of dopaminergic neurons in mice was significantly reduced.The protein phosphorylation of p-PI3K,p-AKT and p-mTOR also increased about 2-fold,compared with the model group.Furthermore,qRT-PCR results demonstrated that the mRNA levels of pro-inflammatory cytokines,IL-1βand TNF-α,were reduced,but the level of anti-inflammatory cytokine IL-10 increased after DMTHB treatment.Finally,the cellular assay displayed that DMTHB was also a strong antioxidant to protect neuron cell line PC12 by scavenging ROS.In this study,we demonstrated DMTHB alleviates the behavioral disorder and protects dopaminergic neurons through multiple-target effects includubg anti-apoptotic,anti-inflammatory and antioxidant effects.
