Buddlejasaponin IV

Phytother Res. 2011 Oct;25(10):1503-10.

Buddlejasaponin IV induces cell cycle arrest at G2/M phase and apoptosis in immortalized human oral keratinocytes.[Pubmed: 21394802]

Buddlejasaponin IV (BS-IV), a major component of Pleurospermum kamtschaticum, exerts antiinflammatory and cytotoxic effects against cancer cells.
METHODS AND RESULTS:
The study investigated whether Buddlejasaponin IV could prevent oral carcinogenesis by inhibiting the growth of immortalized human oral keratinocytes (IHOKs). Buddlejasaponin IV reduced cell viability and induced cell cycle arrest at G2/M phase and apoptotic morphological changes in IHOKs. Buddlejasaponin IV inhibited the levels of cyclin B1, Cdc2 and Cdc25C, but enhanced Chk2 phosphorylation. The increased levels of pRb and p21 protein and the activation of p53 were also noted in Buddlejasaponin IV-treated IHOKs. In addition, Buddlejasaponin IV induced cytochrome c release from mitochondria by reducing antiapoptotic Bcl-2 levels and increasing pro-apoptotic Bax levels. Buddlejasaponin IV treatment resulted in the activation of caspase-9 and caspase-3. PARP cleavage was also clearly observed in the Buddlejasaponin IV-treated IHOKs. Furthermore, the expression of the Fas death receptor and Fas ligand was induced and procaspase-8 level was suppressed by Buddlejasaponin IV treatment. Taken together, Buddlejasaponin IV treatment inhibited the growth of IHOK cells via the induction of p53-dependent cell cycle arrest at the G2/M phase and apoptosis via both mitochondrial-dependent and death receptor-mediated pathways.
CONCLUSIONS:
Thus, Buddlejasaponin IV can be considered an excellent candidate for a chemopreventive agent to block the progression of HPV-induced oral carcinogenesis.

Cancer Res.,2005,46:2464.

Antimetastatic activity of the extract of Pleurospermum kamtschaticum and its active component, Buddlejasaponin IV through the induction of detachment-mediated apoptosis(Anoikis) and the expression of nonsteroidal anti-inflammatory drug-activated gene(NAG[Reference: WebLink]

Buddlejasaponin IV(BS-IV) is one of the active components of the P. kamtschaticum. Apoptosis is the process of programmed cell death or cell suicide. Anoikis is a poorly characterized form of apoptosis that occurs when adherent cells lose their integrin-mediated attachment to the extracellular matrix. Anoikis has been suggested to act as a physiological barrier to metastasis. NAG-1, a TGF-β superfamily member also has antitumorigenic activity and stimulates apoptosis in colon cancer and other cell lines. Cell viability was assessed using a MTT and Trypan Blue exclusion assay. Apoptosis was investigated in terms of DAPI staining, DNA laddering and a flow cytometry analysis. The expression of NAG-1, MAPKs and Bcl-2 family members were assessed by western blotting.
METHODS AND RESULTS:
In this study, we found that the methanolic extract of P. kamtschaticum and BS-IV inhibited the viability of HT-29 cells in a dose-related manner. The extract of P. kamtschaticum and BS-IV induced apoptosis, showing nuclear condensation, DNA fragmentation and the increment of annexin V-FITC-positive fluorescent cells. In particular, the extract of P. kamtschaticum and BS-VI reduced the adhesion of HT-29cells on culture flask and detached the cells within two hours. Moreover the extract of P. kamtschaticum and BS-IV dose-dependently upregulated the expression of NAG-1. Based on these data, to investigate the molecular mechanisms underlying the induction of anoikis and NAG-1 upregulation by the extract of P. kamtschaticum and BS-IV, we assessed the expression and/or activation of integrin-dependent kinases, MAPKs and Bcl-2 family, especially BH3 only proteins. Antimetastatic effect was confirmed in spontaneous mouse lung metastatis model.
CONCLUSIONS:
These results suggest that the extract of P. kamtschaticum and BS-IV may possess antimetastatic potential by inducing anoikis and upregulating NAG-1 expression.

Br J Pharmacol. 2006 May;148(2):216-25.

Anti-inflammatory effect of buddlejasaponin IV through the inhibition of iNOS and COX-2 expression in RAW 264.7 macrophages via the NF-kappaB inactivation.[Pubmed: 16520738]

Buddlejasaponin IV isolated from Pleurospermum kamtschatidum is an anti-inflammatory compound that inhibits NO, PGE(2) and TNF-alpha production.
METHODS AND RESULTS:
Here, we studied the mode of action of this compound. Buddlejasaponin IV (2.5-10 microM) reduced lipopolysaccaride (LPS (1 microg ml(-1)))-induced levels of iNOS and COX-2 at the protein levels, and iNOS, COX-2, TNF-alpha, interleukin (IL)-1beta and IL-6 mRNA expression in RAW 264.7 macrophages in a concentration-dependent manner, as determined by Western blotting and RT-PCR, respectively. Buddlejasaponin IV inhibited the LPS-induced activation of nuclear factor-kappaB (NF-kappaB), a transcription factor necessary for proinflammatory mediators, iNOS, COX-2, TNF-alpha, IL-1beta and IL-6 expression. This effect was accompanied by a parallel reduction in IkappaB-alpha degradation and phosphorylation, and by the nuclear translocation of the NF-kappaB p65 subunit. The effects of Buddlejasaponin IV on acute phase inflammation were studied on serotonin- and carrageenan-induced paw edema. The antiedematous effect of Buddlejasaponin IV was compared with 10 mg kg(-1) of indomethacin p.o. Maximum inhibitions of 26 and 41% were noted at a dose of 20 mg kg(-1) for serotonin- and carrageenan-induced paw edema, respectively. The analgesic effect of Buddlejasaponin IV was evaluated using acetic acid-induced writhing and hot-plate tests. Buddlejasaponin IV (10 and 20 mg kg(-1), p.o.) was found to have a marked analgesic effect in both models.
CONCLUSIONS:
These results suggest that the inhibitions of the expressions of iNOS, COX-2, TNF-alpha, IL-1beta and IL-6 by blocking NF-kappaB activation, are responsible for the anti-inflammatory effects of Buddlejasaponin IV isolated from P. kamtschatidum.

J Cancer Prev. 2013 Dec;18(4):330-6.

The Inhibitory Effect of Buddlejasaponin IV on the Growth of YD-10B Human Oral Squamous Cell Carcinoma Cells.[Pubmed: 25337562]

Buddlejasaponin IV (BS-IV), a triterpene saponin isolated from Pleurospermum kamtschaticum HOFFMANN (Umbelliferae), is known to have potent anti-inflammatory activity and cytotoxicity against diverse cancer cell lines. In the present study, we attempted to verify whether Buddlejasaponin IV could inhibit cell growth, and induce cell cycle arrest and apoptosis in highly invasive YD-10B human oral squamous cell carcinoma (OSCC) cells.
METHODS AND RESULTS:
YD-10B cells were treated with various concentrations of Buddlejasaponin IV, and the cell viability was evaluated by MTT assay. Flow cytometry was conducted to examine cell phase distribution and DAPI staining was performed to observe apoptotic morphological changes in Buddlejasaponin IV-treated YD-10B cells. Western blot analysis was used to investigate the expression of proteins associated with cell cycle arrest and apoptosis. RESULTS: Buddlejasaponin IV treatment significantly reduced the viability of YD-10B cells and partially arrested cell cycle progression at the G2/M phase. Treatment with Buddlejasaponin IV substantially decreased the levels of cyclin B1 and stimulated the phosphorylation of checkpoint kinase 2 (Chk2). The expression of p21 was increased but the phosphorylation of Akt was inhibited in Buddlejasaponin IV-treated YD-10B cells. Furthermore, Buddlejasaponin IV induced release of cytochrome c from mitochondria by reducing anti-apoptotic Bcl-2 level and increasing pro-apoptotic Bax level. Active caspase-3 level and the cleavage of poly (ADP-ribose) polymerase (PARP) were enhanced by Buddlejasaponin IV treatment. In addition, Buddlejasaponin IV increased the expression of Fas death receptor and its ligand (FasL) in YD-10B cells.
CONCLUSIONS:
The treatment with Buddlejasaponin IV inhibits the growth of YD-10B cells by inducing p21-dependent cell cycle arrest at G2/M phase and apoptosis through both mitochondrial-dependent and death receptor-mediated pathways. Thus, Buddlejasaponin IV is an excellent candidate for a chemopreventive agent to block the progression of human OSCC.

J Ethnopharmacol. 2007 Jun 13;112(2):255-61. Epub 2007 Mar 14.

The MeOH extract of Pleurospermum kamtschaticum and its active component buddlejasaponin (IV) inhibits intrinsic and extrinsic hyperlipidemia and hypercholesterolemia in the rat.[Pubmed: 17433587]

METHODS AND RESULTS:
Since the MeOH extract and the BuOH fraction of Pleurospermum kamtschaticum were found to be active using these four hypolipidemic assays, its major saponin Buddlejasaponin IV {BS(IV)} isolated from the BuOH fraction were also tested to demonstrate the active components. Buddlejasaponin IV was found to significantly inhibit hypercholesterolemia and hyperlipidemia by extrinsic and intrinsic inducers. In particular, Buddlejasaponin IV reduced the blood thiobarbituric acid reactive substance (TBARS) and hydroxy radical levels, and increased superoxide dismutase activity in high cholesterol diet-induced rats, thus suggesting that Buddlejasaponin IV reduces oxidative stress caused by a high cholesterol diet. Moreover, these effects of Buddlejasaponin IV were comparable to probucol, which was used as a positive control.
CONCLUSIONS:
These results suggested that Pleurospermum kamtschaticum which is traditionally used to treat atherosclerosis and its active major saponin Buddlejasaponin IV could be used to treat hypercholesterolemia or hyperlipidemia.