BACKGROUND Altered tight junction(TJ)proteins are correlated with carcinogenesis and tumor development.Nimbolide is a tetranotriterpenoid that has been shown to have antioxidant and anti-proliferative properties;howev...BACKGROUND Altered tight junction(TJ)proteins are correlated with carcinogenesis and tumor development.Nimbolide is a tetranotriterpenoid that has been shown to have antioxidant and anti-proliferative properties;however,its anticancer effects and molecular mechanism in hepatocellular carcinoma(HCC)remains obscure.AIM To investigate the effect of nimbolide on TJ proteins,cell cycle progression,and hepatic inflammation in a mouse model of HCC.METHODS HCC was induced in male Swiss albino mice(CD-1 strain)by a single intraperitoneal injection of 100 mg/kg diethylnitrosamine(DEN)followed by 80 ppm N-nitrosomorpholine(NMOR)in drinking water for 28 wk.After 28 wk,nimbolide(6 mg/kg)was given orally for four consecutive weeks in DEN/NMOR induced HCC mice.At the end of the 32nd week,all the mice were sacrificed and blood and liver samples were collected for various analyses.Macroscopic examinations of hepatic nodules were assessed.Liver histology and HCC tumor markers such as alpha-fetoprotein(AFP)and glypican-3 were measured.Expression of TJ proteins,cell proliferation,and cell cycle markers,inflammatory markers,and oxidative stress markers were analyzed.In silico analysis was performed to confirm the binding and modulatory effect of nimbolide on zonula occludens 1(ZO-1),nuclear factor of kappa light polypeptide gene enhancer in B-cells(NF-κB),and tumor necrosis factor alpha(TNF-α).RESULTS We found nimbolide treatment at a concentration of 6 mg/kg to HCC mice reduced hepatic tumor size by 52.08%and tumor volume(P<0.01),and delayed tumor growth in HCC mice with a concomitant reduction in tumor markers such as AFP levels(P<0.01)and glypican-3 expression(P<0.05).Furthermore,nimbolide treatment increased tight junction proteins such as ZO-1 and occludin expression(P<0.05,respectively)and reduced ZO-1 associated nucleic acid binding protein expression(P<0.001)in HCC mice liver.Nimbolide treatment to HCC mice also inhibited cell proliferation and suppressed cell cycle progression by attenuating proliferating cell nuclear antigen(P<0.01),cyclin dependent kinase(P<0.05),and CyclinD1(P<0.05)expression.In addition,nimbolide treatment to HCC mice ameliorated hepatic inflammation by reducing NF-κB,interleukin 1 beta and TNF-αexpression(P<0.05,respectively)and abrogated oxidative stress by attenuating 4-hydroxynonenal expression(P<0.01).Molecular docking studies further confirmed that nimbolide interacts with ZO-1,NF-κB,and TNF-α.CONCLUSION Our current study showed for the first time that nimbolide exhibits anticancer effect by reducing tumor size,tumor burden and by suppressing cell cycle progression in HCC mice.Furthermore,nimbolide treatment to HCC mice ameliorated inflammation and oxidative stress,and improved TJ proteins expression.Consequently,nimbolide could be potentially used as a natural therapeutic agent for HCC treatment,however further human studies are warranted.展开更多
OBJECTIVE Prostate cancer is one of the most commonly diagnosed cancers worldwide.Current therapies for metastatic prostate cancer are only marginally effective,and hence novel treatment modalities are urgently needed...OBJECTIVE Prostate cancer is one of the most commonly diagnosed cancers worldwide.Current therapies for metastatic prostate cancer are only marginally effective,and hence novel treatment modalities are urgently needed.Considerable evidence suggests that chronic inflammation plays a pivotal role in the development and progression of prostate cancer.Thus agents that can suppress these inflammatory mediators could form the basis of novel therapy for prostate cancer patients.In our study,we focused on analyzing the potential anticancer effects of nimbolide,a terpenoid lactone derived from Azadirachta indica(Neem tree)against prostate cancer.METHODS Molecular biology techniques such as western blot analysis,DNA binding,luciferase assays,and immunohistochemistry were used for both in vitro and in vivo experiments.RESULTS Data from the in vitrostudies indicated that nimbolide could inhibit cell proliferation,induce apoptosis and suppress cellular invasion and migration.Interestingly,nimbolide also abrogated the activation of pro-inflammatory STAT 3 transcription factor,and this effect was found to be mediated via an increased production of reactive oxygen species(ROS),whereas depletion of ROS attenuated pSTAT 3 inhibitory effects of the drug.The in vivo efficacy of nimbolide was also noted in transgenic adenocarcinoma of mouse prostate(TRAMP)model,in which this triterpenoid significantly suppressed the tumor progression and growth without exhibiting any substantial adverse effects.CONCLUSION Overall our findings indicate that nimbolide exhibits significant anticancer effects in prostate cancer,and these effects may be mediated at least in part through the modulation of STAT 3 signaling pathway.展开更多
Inflammation,abnormal cholesterol metabolism,and macrophage infiltration are involved in the destruction of the extracellular matrix of the nucleus pulposus(NP),culminating in intervertebral disc degeneration(IDD).Whe...Inflammation,abnormal cholesterol metabolism,and macrophage infiltration are involved in the destruction of the extracellular matrix of the nucleus pulposus(NP),culminating in intervertebral disc degeneration(IDD).Whether nimbolide(Nim),a natural extract,can alleviate IDD is unclear.In this study,we demonstrated that Nim promotes cholesterol efflux and inhibits the activation of the nuclear factor kappa B(NF-κB)and mitogen-activated protein kinase(MAPK)signaling pathways by activating sirtuin 1(SIRT1)in nucleus pulposus cells(NPCs)during inflammation.Thus,Nim balanced matrix anabolism and catabolism of NPCs.However,the inhibition of SIRT1 significantly attenuated the effects of Nim.We also found that Nim promoted the expression of SIRT1 in RAW 264.7,which enhanced the proportion of M2 macrophages by facilitating cholesterol homeostasis reprogramming and impeded M1-like macrophages polarization by blocking the activation of inflammatory signaling.Based on these results,Nim can improve the microenvironment and facilitate matrix metabolism equilibrium in NPCs.Furthermore,in vivo treatment with Nim delayed IDD progression by boosting SIRT1 expression,modulating macrophage polarization and preserving the extracellular matrix.In conclusion,Nim may represent a novel therapeutic strategy for treating IDD.展开更多
基金Supported by JIPMER intramural research grantIndian Council of Medical Research(ICMR),New Delhi,India,No.3/1/3 J.R.F.-2016/LS/HRDDepartment of Biotechnology,Government of India,No.102/IFD/SAN/22/2013-14.
文摘BACKGROUND Altered tight junction(TJ)proteins are correlated with carcinogenesis and tumor development.Nimbolide is a tetranotriterpenoid that has been shown to have antioxidant and anti-proliferative properties;however,its anticancer effects and molecular mechanism in hepatocellular carcinoma(HCC)remains obscure.AIM To investigate the effect of nimbolide on TJ proteins,cell cycle progression,and hepatic inflammation in a mouse model of HCC.METHODS HCC was induced in male Swiss albino mice(CD-1 strain)by a single intraperitoneal injection of 100 mg/kg diethylnitrosamine(DEN)followed by 80 ppm N-nitrosomorpholine(NMOR)in drinking water for 28 wk.After 28 wk,nimbolide(6 mg/kg)was given orally for four consecutive weeks in DEN/NMOR induced HCC mice.At the end of the 32nd week,all the mice were sacrificed and blood and liver samples were collected for various analyses.Macroscopic examinations of hepatic nodules were assessed.Liver histology and HCC tumor markers such as alpha-fetoprotein(AFP)and glypican-3 were measured.Expression of TJ proteins,cell proliferation,and cell cycle markers,inflammatory markers,and oxidative stress markers were analyzed.In silico analysis was performed to confirm the binding and modulatory effect of nimbolide on zonula occludens 1(ZO-1),nuclear factor of kappa light polypeptide gene enhancer in B-cells(NF-κB),and tumor necrosis factor alpha(TNF-α).RESULTS We found nimbolide treatment at a concentration of 6 mg/kg to HCC mice reduced hepatic tumor size by 52.08%and tumor volume(P<0.01),and delayed tumor growth in HCC mice with a concomitant reduction in tumor markers such as AFP levels(P<0.01)and glypican-3 expression(P<0.05).Furthermore,nimbolide treatment increased tight junction proteins such as ZO-1 and occludin expression(P<0.05,respectively)and reduced ZO-1 associated nucleic acid binding protein expression(P<0.001)in HCC mice liver.Nimbolide treatment to HCC mice also inhibited cell proliferation and suppressed cell cycle progression by attenuating proliferating cell nuclear antigen(P<0.01),cyclin dependent kinase(P<0.05),and CyclinD1(P<0.05)expression.In addition,nimbolide treatment to HCC mice ameliorated hepatic inflammation by reducing NF-κB,interleukin 1 beta and TNF-αexpression(P<0.05,respectively)and abrogated oxidative stress by attenuating 4-hydroxynonenal expression(P<0.01).Molecular docking studies further confirmed that nimbolide interacts with ZO-1,NF-κB,and TNF-α.CONCLUSION Our current study showed for the first time that nimbolide exhibits anticancer effect by reducing tumor size,tumor burden and by suppressing cell cycle progression in HCC mice.Furthermore,nimbolide treatment to HCC mice ameliorated inflammation and oxidative stress,and improved TJ proteins expression.Consequently,nimbolide could be potentially used as a natural therapeutic agent for HCC treatment,however further human studies are warranted.
基金The project supported by agrant from NUS Academic Research Fund
文摘OBJECTIVE Prostate cancer is one of the most commonly diagnosed cancers worldwide.Current therapies for metastatic prostate cancer are only marginally effective,and hence novel treatment modalities are urgently needed.Considerable evidence suggests that chronic inflammation plays a pivotal role in the development and progression of prostate cancer.Thus agents that can suppress these inflammatory mediators could form the basis of novel therapy for prostate cancer patients.In our study,we focused on analyzing the potential anticancer effects of nimbolide,a terpenoid lactone derived from Azadirachta indica(Neem tree)against prostate cancer.METHODS Molecular biology techniques such as western blot analysis,DNA binding,luciferase assays,and immunohistochemistry were used for both in vitro and in vivo experiments.RESULTS Data from the in vitrostudies indicated that nimbolide could inhibit cell proliferation,induce apoptosis and suppress cellular invasion and migration.Interestingly,nimbolide also abrogated the activation of pro-inflammatory STAT 3 transcription factor,and this effect was found to be mediated via an increased production of reactive oxygen species(ROS),whereas depletion of ROS attenuated pSTAT 3 inhibitory effects of the drug.The in vivo efficacy of nimbolide was also noted in transgenic adenocarcinoma of mouse prostate(TRAMP)model,in which this triterpenoid significantly suppressed the tumor progression and growth without exhibiting any substantial adverse effects.CONCLUSION Overall our findings indicate that nimbolide exhibits significant anticancer effects in prostate cancer,and these effects may be mediated at least in part through the modulation of STAT 3 signaling pathway.
基金Center of Pharmaceutical Technology of Tsinghua University(Beijing,China)for the support of Schrodinger software and molecular dockingthe National Natural Science Foundation of China(grant numbers 91849114,82030068,and 82172506)。
文摘Inflammation,abnormal cholesterol metabolism,and macrophage infiltration are involved in the destruction of the extracellular matrix of the nucleus pulposus(NP),culminating in intervertebral disc degeneration(IDD).Whether nimbolide(Nim),a natural extract,can alleviate IDD is unclear.In this study,we demonstrated that Nim promotes cholesterol efflux and inhibits the activation of the nuclear factor kappa B(NF-κB)and mitogen-activated protein kinase(MAPK)signaling pathways by activating sirtuin 1(SIRT1)in nucleus pulposus cells(NPCs)during inflammation.Thus,Nim balanced matrix anabolism and catabolism of NPCs.However,the inhibition of SIRT1 significantly attenuated the effects of Nim.We also found that Nim promoted the expression of SIRT1 in RAW 264.7,which enhanced the proportion of M2 macrophages by facilitating cholesterol homeostasis reprogramming and impeded M1-like macrophages polarization by blocking the activation of inflammatory signaling.Based on these results,Nim can improve the microenvironment and facilitate matrix metabolism equilibrium in NPCs.Furthermore,in vivo treatment with Nim delayed IDD progression by boosting SIRT1 expression,modulating macrophage polarization and preserving the extracellular matrix.In conclusion,Nim may represent a novel therapeutic strategy for treating IDD.
