Nitidine chloride

J Ethnopharmacol. 2012 Oct 31;144(1):145-50.

Nitidine chloride inhibits LPS-induced inflammatory cytokines production via MAPK and NF-kappaB pathway in RAW 264.7 cells.[Pubmed: 22971898 ]

Zanthoxylum nitidium (Roxb.) DC. has long been used as a traditional herbal medicine for inflammatory diseases such as rheumatic arthritis and peridentitis. However, the anti-inflammatory mechanism of Nitidine chloride has not been fully elucidated. To determine the anti-inflammatory effects and mechanism of Nitidine chloride (NTD), a pentacyclic alkaloid is isolated from the root of Z. nitidium, in murine macrophages.
METHODS AND RESULTS:
Anti-inflammatory properties of NTD were investigated using lipopolysaccharide (LPS)-stimulated Raw 264.7 macrophages as in vitro model. The pro-inflammatory cytokines were evaluated by real-time RT-PCR and ELISA. Furthermore, intracellular signaling pathways were analyzed by Western blot and Immunofluorescence staining using specific antibodies. NTD significantly reduced the production of pro-inflammatory cytokines such as tumor necrosis factor alpha (TNF-α), interleukin-1β (IL-1β), and IL-6 in both RNA and protein level. Moreover, transcriptional activity of NF-кB as well as the phosphorylation of mitogen-activated protein kinases (MAPKs) in LPS-treated RAW 264.7 was significantly inhibited by NTD in a dose dependent manner. These results suggested that NTD exerts an anti-inflammatory property by inhibiting TNF-α, IL-1β, and IL-6 production in association with reduced NF-κB and MAPK signaling pathways in RAW 264.7 cells.
CONCLUSIONS:
These results suggested that NTD exerts an anti-inflammatory property by inhibiting TNF-α, IL-1β, and IL-6 production in association with reduced NF-κB and MAPK signaling pathways in RAW 264.7 cells. Nitidine chloride inhibits LPS-induced TNF alpha, IL-1beta and IL-6 production via the suppression of phosphorylation of MAPK and the translocation of p65. In addition, these results revealed a novel role of NTD in regulation of inflammatory diseases.

Food Chem Toxicol. 2013 Oct;60:246-51.

Nitidine chloride inhibits renal cancer cell metastasis via suppressing AKT signaling pathway.[Pubmed: 23911800]

Nitidine chloride (NC) has been shown to have anti-cancer effects on various tumors. However, whether NC could exert anti-metastasis activity in renal cancer cells and the underlying mechanisms have not been elucidated.
METHODS AND RESULTS:
In this work, our data demonstrated the anti-metastasis effects of NC on renal cancer cells in vitro. With scratch assay and transwell assays, we found that NC potently suppressed the migration and invasion of 786-O and A498 cells. Mechanistically, we presented that NC significantly decreased phosphorylation of AKT, accompanied by down-regulation of MMP-2 and MMP-9. Furthermore, a specific AKT inhibitor, LY294002, could enhance the anti-metastasis effects of NC, which indicated that NC suppressed metastasis of renal cancer cells partly via inhibition of AKT activity.
CONCLUSIONS:
Taken together, our results imply that NC can be developed as a potential anti-metastasis agent to renal cancer.

Tumour Biol. 2014 Oct;35(10):10201-12.

Nitidine chloride induces apoptosis, cell cycle arrest, and synergistic cytotoxicity with doxorubicin in breast cancer cells.[Pubmed: 25027404]

Medicinal plant extracts have been widely used for cancer treatment. Nitidine chloride (NC) is a natural bioactive alkaloid that has recently been reported to have diverse anticancer properties. We aimed to investigate the cytotoxic effects of NC and the effectiveness of combinatorial treatment including NC and doxorubicin in breast cancer cells.
METHODS AND RESULTS:
Using MTT and flowcytometry assays, we found that NC induced cell growth inhibition and G2/M cell cycle arrest in a time- and dose-dependent manner both in MCF-7 and MDA-MB-231 breast cancer cell lines. Cancer cell growth inhibition was associated with increased levels of the p53 and p21 proteins. Apoptosis induction by NC treatment was confirmed by JC-1 mitochondrial membrane potential, annexin V-positive cell, and TUNEL staining. Using western blot analysis, we found that NC upregulated the pro-apoptotic proteins Bax, cleaved caspase-9 and -3 and cleaved PARP and that it downregulated the anti-apoptotic proteins Bcl-2 and PARP. By using the PI3K/Akt inhibitor LY294002, we further demonstrated that NC-induced apoptosis might be Akt-specific or dependent. In addition, NC exhibited a synergistic effect with doxorubicin on the growth inhibition of the human breast cancer cell lines MCF-7 and MDA-MB-231.
CONCLUSIONS:
Our study demonstrated the anticancer effect of NC on breast cancer and highlighted the potential clinical application of NC.

Oncol Rep. 2015 Mar;33(3):1264-74.

Nitidine chloride induces apoptosis in human hepatocellular carcinoma cells through a pathway involving p53, p21, Bax and Bcl-2.[Pubmed: 25530218]

Nitidine chloride (NC), a novel benzo[c]phenanthridine alkaloid, induces the growth inhibition of cancer cells. Previously it was demonstrated that SMMC-7721 human hepatocellular carcinoma (HCC) cells are highly susceptible to the antiproliferative effects of NC. However, the specific mechanisms remained unclear. In the present study the pathways of growth inhibition induced by NC in SMMC-7721 cells were investigated.
METHODS AND RESULTS:
The effects of NC on SMMC-7721 cell proliferation were characterized by MTT and colony formation assays. Additionally, BALB/c nude mice were transplanted with SMMC-7721 cells to verify the inhibition of HCC by NC in vivo. The results showed that NC inhibited the proliferation of SMMC-7721 cells in vitro in a time- and dose-dependent manner and identified efficacy in vivo in a mouse model of HCC. Acridine orange (AO) staining, transmission electron microscopy, Annexin V/PI staining, TUNEL assay and caspase-3 activation assays were used to investigate apoptosis and the cell cycle distribution. Inhibition was mediated in part by cell cycle arrest in G2/M, leading to chromatin condensation, DNA fragmentation and the formation of apoptotic bodies. Apoptosis was also verified by Annexin V/PI staining, TUNEL assay and caspase-3 activation. To assess the levels of the cell cycle and apoptotic regulators, immunohistochemical staining, ELISA, real-time PCR and RNA interference (RNAi) were employed. The apoptotic process triggered by NC involved the upregulation of p53, p21 and Bax, and the downregulation of Bcl-2.
CONCLUSIONS:
These data elucidate a pathway of apoptosis in SMMC‑7721 cells that involves G2/M arrest, upregulation of p53, Bax, caspase-3 and p21, and downregulation of Bcl-2.

Chinese Journal of Clinical Rehabilitation, 2006, 10(27):171-4.

Protective effects of nitidine chloride on rats during myocardial ischemia/reperfusion.[Reference: WebLink]

①1 and 2 mg/kg Nitidine chloride can reduce the incidence rate of cardiac arrhythmia in rats with myocardial ischemia and reperfusion, postpone the emergence time of cardiac arrhythmia and shorten its duration, decrease the degree of ST segment elevation after reperfusion for 15 minutes and 60 minutes, which have similar effect with verapamil. ②1 and 2 mg/kg Nitidine chloride can reduce the release of myocardial enzyme, relieve the severity of oxygen-derived free radicals injury, and has the effect of protecting myocardial injury during ischemia-reperfusion, in which represents a dose-dependent effect.