The H2S donor sulforaphane inhibits NLRP3 inflammasome activation by inducing mitochondrial autophagy and mitigating CBS-H2S axis damage in in-vitro and in-vivo models of Parkinson's disease.

Sulforaphane attenuates activation of NLRP3 and NLRC4 inflammasomes but not AIM2 inflammasome

Neuroprotective effect of sulforaphane in 6-hydroxydopamine-lesioned mouse model of Parkinson's disease

Sulforaphane, an NRF2 agonist, alleviates ferroptosis in acute liver failure by regulating HDAC6 activity

We investigated the effects of sulforaphane (SFN), an isothiocyanate from cruciferous vegetables, in the regulation of cerebral blood flow using cranial windows in newborn pigs. SFN administered topically (10 µM–1 mM) or systemically (0.4 mg/kg ip) caused immediate and sustained dilation of pial arterioles concomitantly with elevated H2S in periarachnoid cortical cerebrospinal fluid. H2S is a potent vasodilator of cerebral arterioles. SFN is not a H2S donor but it acts via stimulating H2S generation in the brain catalyzed by cystathionine γ-lyase (CSE) and cystathionine β-synthase (CBS). CSE/CBS inhibitors propargylglycine, β-cyano-L-alanine, and aminooxyacetic acid blocked brain H2S generation and cerebral vasodilation caused by SFN. The SFN-elicited vasodilation requires activation of potassium channels in cerebral arterioles. The inhibitors of KATP and BK channels glibenclamide, paxilline, and iberiotoxin blocked the vasodilator effects of topical and systemic SFN, supporting the concept that H2S is the mediator of the vasodilator properties of SFN in cerebral circulation. Overall, we provide first evidence that SFN is a brain permeable compound that increases cerebral blood flow via a non-genomic mechanism that is mediated via activation of CSE/CBS-catalyzed H2S formation in neurovascular cells followed by H2S-induced activation of KATP and BK channels in arteriolar smooth muscle.

Characterization and neurotherapeutic evaluation of venom polypeptides identified from Vespa magnifica: The role of Mastoparan-M in Parkinson's disease intervention.

Deferoxamine-mediated up-regulation of HIF-1α prevents dopaminergic neuronal death via the activation of MAPK family proteins in MPTP-treated mice

Methamphetamine (METH) is a powerfully addictive stimulant associated with serious health conditions. Accumulating evidence suggests a role of oxidative stress in METH-induced behavioral abnormalities. Sulforaphane (SFN), found in cruciferous vegetables, is a potent antioxidant. It is of interest to determine whether SFN can attenuate behavioral and neuropathological changes associated with METH exposure. This study was undertaken to examine the effects of SFN on behavioral changes and dopaminergic neurotoxicity in mice exposed to METH. The effects of SFN on acute hyperlocomotion and the development of behavioral sensitization induced by the administration of METH were examined. Levels of dopamine (DA) and its major metabolite 3,4-dihydroxyphenyl acetic acid (DOPAC) in the striatum were measured. In addition, DA transporter (DAT) immunoreactivity was also performed. Pretreatment with SFN at 1, 3, and 10mg/kg elicited a dose-dependent attenuation of acute hyperlocomotion in mice, after a single administration of METH (3mg/kg). The development of behavioral sensitization after repeated administrations of METH (3mg/kg/day, once daily for 5days) was significantly reduced by pretreatment with SFN (10mg/kg). In addition, the lowering of DA levels and DOPAC as well as DAT immunoreactivity in the striatum, usually seen after repeated administration of METH, was significantly attenuated by both pretreatment and the subsequent administration of SFN. Furthermore, SFN significantly reduced microglial activation in the striatum after repeated exposure to METH. It is therefore likely that SFN can be a useful drug for the treatment of signs associated with METH abuse in humans.

ObjectiveAccumulating evidence suggests that oxidative stress plays a role in the pathophysiology of schizophrenia and that the potent antioxidants may be potential therapeutic drugs for schizophrenia. This study was undertaken to examine the effects of the potent antioxidant sulforaphane (SFN), found in cruciferous vegetables, on behavioral abnormalities (e.g., hyperlocomotion and prepulse inhibition [PPI] deficits) in mice after a single administration of the N-methyl-D-aspartate (NMDA)-receptor antagonist phencyclidine (PCP).MethodsEffects of SFN (3, 10, and 30 mg/kg, intraperitoneally [i.p.]) on hyperlocomotion and PPI deficits in the adult male ddY mice after administration of PCP (3.0 mg/kg, subcutaneously [s.c.]) were examined.ResultsAdministration of SFN (30 mg/kg, intraperitoneally [i.p.]), but not low doses (3 and 10 mg/kg, i.p.), significantly attenuated hyperlocomotion in mice after PCP administration (3.0 mg/kg, subcutaneously [s.c.]). Furthermore, administration of SFN (3, 10, and 30 mg/kg, i.p.) attenuated the PPI deficits in mice after PCP administration (3.0 mg/kg, s.c.) in a dose-dependent manner.ConclusionThese results suggest that SFN has antipsychotic activity in an animal model of schizophrenia. Therefore, it is likely that SFN may be a potential therapeutic drug for schizophrenia.

Neuroinflammation, particularly through the NLRP3 inflammasome, plays a significant role in the progression of Parkinson's disease (PD). Icaritin (ICT), a flavonoid compound derived from Epimedium, has demonstrated potential therapeutic effects on PD. In this study, an MPTP-induced PD mouse model was used to evaluate the impact of ICT. The mice were treated with ICT through intragastric administration, and their motor functions were assessed by the rotarod test and the pole test. Immunofluorescence was used to examine the expression of TH, Iba1, and GFAP, and Western blotting was used to measure changes in the levels of NLRP3 and other relevant proteins. The results showed that ICT significantly improved motor dysfunction induced by MPTP, reduced dopaminergic neuronal damage, diminished glial cell activation, and inhibited the expression of proteins in the NLRP3 signalling pathway. These findings suggest that ICT has a protective effect on the MPTP-induced PD mouse model and that its mechanism of action is likely related to the inhibition of NLRP3-mediated neuroinflammation.

RND3 modulates microglial polarization and alleviates neuroinflammation in Parkinson's disease by suppressing NLRP3 inflammasome activation

Organophosphorus flame retardant TDCPP, a substitute for brominated flame retardants, is widely used in consumer products but readily leaches into the environment, posing human exposure risks. This study investigated the neurotoxic mechanisms of TDCPP in Parkinson's disease (PD). Using an MPTP-induced PD mouse model, TDCPP exposure exacerbated behavioral deficits, reduced tyrosine hydroxylase (TH)-positive neurons in the substantia nigra, and amplified neuroinflammation characterized by enhanced microglial reactivity, elevated pro-inflammatory IFN-γ, and diminished anti-inflammatory IL-4 and regulatory T cells (Tregs). Concurrently, TDCPP lowered glutathione (GSH) levels and altered ferroptosis-related protein expression, indicating oxidative stress involvement. In SH-SY5Y cells, co-treatment with TDCPP and MPTP caused mitochondrial membrane depolarization, increased reactive oxygen species (ROS), and shifted microglia into a pro-inflammatory state-evidenced by increased CD86 expression-and impaired their phagocytic clearance of α-synuclein. These findings demonstrate that TDCPP aggravates PD neurodegeneration through dual mechanisms: ferroptosis-linked oxidative stress and neuroinflammation. This study provides the first evidence linking TDCPP exposure to ferroptosis-mediated neurotoxicity in PD models, bridging environmental toxicology and neurodegenerative research. The results underscore the role of environmental pollutants in PD progression and offer critical insights for refining safety regulations to mitigate human health risks.

Background: Sulforaphane (SFN), a phytochemical compound, which belongs to isothiocyanates family found in abundance in broccoli sprouts, potently induces a variety of antioxidant enzymes via nrf2-keap1 mediated pathway, thereby protects cells from injury induced by various kinds of oxidative stresses. We have previously shown that SFN protects gastric mucosa from oxidative injury induced by H. pylori infection. SFN also down-regulates histone deacetylase (HDAC) activity, thereby induces apoptosis and inhibits proliferation of tumor cells in variety of tissues. On the other hand, colon cancer has been increasing in Japan. Since numerous epidemiological studies have shown that colon cancer is inversely associated with intake of anti-oxidant vegetables, we examined if dietary SGS prevents colon tumorigenesis in mice, and in human subjects.Methods: 1. Effects of SFN on Colonic tumorigenesis in Mice Treated with Chemical Carcinogen: Effects of SFN on colonic tumorigenesis were examined in the ICR male mice, pretreated with a chemical carcinogen, azoxymethane (AOM) (15 mg/kg). The AOM-treated mice were fed for 8 weeks with or without sulforaphane glucosinolates (SGS: 2,200 ppm/kg/day), which is a precursor of SFN. Effects of SGS treatment on formation of the microscopic aberrant crypt foci (ACF), and the macroscopic tumors in colonic mucosa were evaluated. 2. Effects of SFN on formation of Colonic Aberrant Crypt Foci (ACF) in patients with colonic adenoma: Effects of intake of raw broccoli sprouts (BS), 50 g/day, which contains 220 mg SGS every other day, for 6 months on changes in the number of ACF in rectal mucosa was examined by colonoscopy in patients with colonic adenoma. 3. Effects of SFN on intestinal microbiota in human subjects: Effects of dietary intake of raw broccoli sprouts (BS), 20 g/day, which contains 88 mg SGS every other day, for 2 weeks on intestinal microbiota in healthy volunteers was assessed by measuring composition of stool bacteria, using T-RFLP method. In human studies, alfalfa sprouts, which contains no SFN was used as a placebo control. Results: 1. Daily administration of SGS suppressed formation of microscopic ACF and macroscopic colonic tumors in the AOM-pretreated mice in vivo. 2. Intake of BS for 6 months tended to decrease the number of colonic ACF in patients with colonic adenoma. 3. Intake of BS for 2 weeks increased percentages of Bifidobacterium and Clostridium XVIa, which has been shown to enhance protection of colonic mucosa by increasing butyrate production in colonic lumen.Conclusions: 1. SFN affords chemoprotection against colonic cancer, presumably by up-regulating nrf2-dependent antioxidant enzymes in normal colonic epithelial cells, and by inducing apoptosis of colonic tumor cells through inhibition of HDAC activity. The present study further suggests that changes in intestinal microbiota by SFN, may play a role in chemoprevention against colon cancer.Keywords: sulforaphane, colon cancer, chemoprevention, aberrant crypt foci, intestinal microbiota

We have shown previously that sulforaphane (SFN), a constituent of many edible cruciferous vegetables including broccoli, suppresses growth of prostate cancer cells in culture as well as in vivo by causing apoptosis, but the sequence of events leading to cell death is poorly defined. Using PC-3 and DU145 human prostate cancer cells as a model, we now demonstrate, for the first time, that the initial signal for SFN-induced apoptosis is derived from reactive oxygen species (ROS). Exposure of PC-3 cells to growth-suppressive concentrations of SFN resulted in ROS generation, which was accompanied by disruption of mitochondrial membrane potential, cytosolic release of cytochrome c, and apoptosis. All these effects were significantly blocked on pretreatment with N-acetylcysteine and overexpression of catalase. The SFN-induced ROS generation was significantly attenuated on pretreatment with mitochondrial respiratory chain complex I inhibitors, including diphenyleneiodonium chloride and rotenone. SFN treatment also caused a rapid and significant depletion of GSH levels. Collectively, these observations indicate that SFN-induced ROS generation is probably mediated by a nonmitochondrial mechanism involving GSH depletion as well as a mitochondrial component. Ectopic expression of Bcl-xL, but not Bcl-2, in PC-3 cells offered significant protection against the cell death caused by SFN. In addition, SFN treatment resulted in an increase in the level of Fas, activation of caspase-8, and cleavage of Bid. Furthermore, SV40-immortalized mouse embryonic fibroblasts (MEFs) derived from Bid knock-out mice displayed significant resistance toward SFN-induced apoptosis compared with wild-type MEFs. In conclusion, the results of the present study indicate that SFN-induced apoptosis in prostate cancer cells is initiated by ROS generation and that both intrinsic and extrinsic caspase cascades contribute to the cell death caused by this highly promising cancer chemopreventive agent.

BackgroundSulforaphane (SFN), a phytochemical compound, which belongs to isothiocyanates family found in abundance in broccoli sprouts, potently induces a variety of antioxidant enzymes via NF‐E2‐related factor 2‐Kelch‐like ECH‐associated protein 1‐mediated pathway, thereby protects cells from injury induced by various kinds of oxidative stresses. We have previously shown that SFN protects gastric mucosa from oxidative injury induced by H. pyloriinfection. SFN also down‐regulates histone deacetylase (HDAC) activity, thereby induces apoptosis and inhibits proliferation of tumor cells in variety of tissues. On the other hand, colon cancer has been increasing in Japan. Since numerous epidemiological studies have shown that colon cancer is inversely associated with intake of anti‐oxidant vegetables, we examined if daily intake of SFN prevents colon tumorigenesis in mice, and in human subjects.Methods1. Effects of SFN on Colonic tumorigenesis in Mice Treated with Chemical Carcinogen: Effects of SFN on colonic tumorigenesis were examined in the ICR male mice, pretreated with a chemical carcinogen, azoxymethane (AOM) (15 mg/kg). The mice treated with AOM for 3 or 6 times were fed for 8 or 24 weeks with or without sulforaphane glucosinolates (SGS: 2,200 ppm/kg/day), which is a precursor of SFN. Effects of SGS treatment on formation of the microscopic aberrant crypt foci (ACF), and the macroscopic tumors in colonic mucosa were evaluated. 2. Effects of SFN on formation of Colonic ACF in patients with colonic adenoma: Effects of intake of raw broccoli sprouts (BS), 50 g/day, which contains 220 mg SGS every other day, for 6 months on changes in the number of ACF in rectal mucosa was examined by colonoscopy in patients with colonic adenoma. 3. Effects of SFN on intestinal microbiota in human subjects: Effects of dietary intake of raw BS, 20 g/day, which contains 88 mg SGS every other day, for 2 weeks on intestinal microbiota in healthy volunteers was assessed by measuring composition of stool bacteria, using a method of terminal restriction fragment length polymorphism flora analysis. In human studies, alfalfa sprouts, which contains no SFN was used as a placebo control.Results1. Daily administration of SGS suppressed formation of microscopic ACF and macroscopic colonic tumors in the AOM‐pretreated mice in vivo. 2. Intake of BS for 6 months tended to decrease the number of colonic ACF in patients with colonic adenoma. 3. Intake of BS for 2 weeks increased percentages of Bifidobacterium and Clostridium XVIa, which has been shown to enhance protection of colonic mucosa by increasing butyrate production in colonic lumen.Conclusions1. Daily intake of SFN affords chemoprotection against colonic tumors in the mice treated with a chemical carcinogen. 2. The present study further suggests that, in addition to previously reported mechanisms, changes in the intestinal microbiota by SFN intake may also play a role in chemoprevention against colon cancer. Further studies are required to prove this possibility.Support or Funding InformationThis study was supported in part by Murakami Farm Inc., Japan.

Background: Sulforaphane (SUL), a kind of isothiocyanate, has been reported to exert strong inhibitory effects on various cancer cells by inducing apoptosis and/or cell cycle arrest. Recently autophagy induction by cancer cells has been focused as a defense mechanism against various proapoptotic agents, which may result in resistance to cancer therapy. Therefore, inhibition of autophagy in cancer cells may be a novel strategy in cancer therapy. Tumor cells depend on angiogenesis for their growth and metastasis and, therefore, anti-angiogenic therapy is a promising anti-cancer strategy. In the present study, we aimed to investigate the anti-angiogenic effect of SUL, and the possible induction of autophagy by tumor vascular endothelial cells as a defense mechanism against SUL-induced apoptosis. Additionally, we investigated the role of specific autophagy inhibitor in potentiating the proapoptotic effect of SUL. For these purposes, the human umbilical vein endothelial cells (HUVECs) were used as the in vitro model of tumor endothelium. Methods: HUVECs were isolated from umbilical cords, obtained from normal pregnants at the delivery, from whom informed consent was obtained. Cells were treated with SUL at different concentrations, and their proliferative activity was assessed by the MTS assay. Apoptosis was investigated by flow-cytometry after double-staining with FITC-conjugated Annexin-V and propidium iodine. The formation of acidic vesicular organelles (AVOs) was detected by fluorescence microscopy and flow-cytometry following staining with acridine orange. Detection of light chain 3 (LC3) levels was investigated by Western blot. Localizations of LC3 and cytochrome c were analyzed by immunocytochemistry. Finally, the vascular-like structure formation on Matrigel was investigated. The role of the specific autophagy inhibitor (3-methyladenine (3-MA)) in potentiating the effects of SUL was similarly investigated. Results: The proapoptotic effect was observed by treatment of cells with relatively high concentrations of SUL for long periods of time. At 16 hour-treatment with lower concentrations of SUL, the features of autophagy, were observed in HUVECs. Treatment of cells with 3-MA potentiated the proapoptotic effect of SUL, which was dependent on the activation of caspases and the release of cytochrome c to the cytosol. SUL dose-dependently inhibited the tube-like formation by HUVECs on Matrigel, an important event in the process of angiogenesis, and treatment of 3-MA potentiated this effect. Conclusion: The present results demonstrate the induction of autophagy in endothelial cells as a protective reaction against the proapoptotic effect of SUL, and consequently, the potentiation of the proapoptotic effect by the inhibition of autophagy. These findings open premises for the use of autophagy inhibitors in combination with chemotherapeutic agents for the anti-angiogenic agents. Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4841.