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Aims: To further investigate the anti-colorectal carcinoma (CRC) effect of Sophoridine (SRI) which is a quinolizidine alkaloid extracted from a traditional Chinese medicine (TCM) Sophora alopecuroides L. and detect the mechanism involved, provide some basis for the development of S. alopecuroides L., Main Methods: The anti-proliferation of SRI in human colorectal cells SW480 were detected by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay. The potential mechanism of anti-proliferation was also investigated using apoptosis assays. The rate of apoptosis cells was detected also. The apoptosis-related proteins cysteinyl aspartate specific protease (caspase), caspase-3, caspase-7, caspase-9, and poly-ADP-ribose-poly-merase (PARP) were determined by western blotting analysis. In animal studies, nude mice were subcutaneously injected with SW480 cells in the armpit to establish the xenograft tumors and administrated with different drugs (control, 5-Fu, SRI H, and SRI L). The general state of health of the mice and the growth of tumors were observed and the inhibitory rate was calculated. The pathology and ultrastructure of xenograft tumors treated with SRI were observed also., Key Findings: SRI significantly inhibited the growth of SW480 cells, and the administration of SRI significantly inhibited the growth of xenograft tumors without apparent toxicity. SRI's mechanism of action involved the induction of apoptosis., Significance: These results suggest that SRI produces obvious anti-tumor effects in vitro and in vivo. It supports the viability of developing SRI as a novel therapeutic prodrug for CRC treatment, as well as providing a method for identifying new anti-tumor drugs in TCM., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Introduction: Sophora alopecuroides L., a traditional Chinese herbal remedy, has been widely used for treating enteritis and bacillary dysentery for many years. Sophocarpine is a major ingredient of S. alopecuroides L. and has a wide range of pharmacological effects., Materials and Methods: In this study, we investigated the therapeutic potential of sophocarpine for treating dextran sulfate sodium (DSS)-induced experimental ulcerative colitis in C57BL/6 mice, a well-characterized murine model of ulcerative colitis. Experimental colitis was induced in these mice by dissolving 5% DSS in their drinking water for 7 days and sophocarpine (60, 30, and 15 mg/kg of body weight) and sulfasalazine (520 mg/kg) were administered orally once a day for 7 days., Results: Sophocarpine significantly ameliorated DSS-induced colitis as identified by a reduced disease activity index and wet weight of colons as well as recovery of body weight. Furthermore, the oral administration of sophocarpine significantly decreased myeloperoxidase activity and the level of interleukin (IL)-1 and IL-6 in serum (P < 0.01), while there was no significant effect on the level of IL-4., Conclusions: In conclusion, sophocarpine significantly ameliorated DSS-induced colitis in mice by regulating the pro- and anti-inflammatory cytokine production. Based upon our results, we suggest that sophocarpine is an effective agent for treating colonic inflammation.

Objective: To investigate the effect of total alkaloids of Sophora alopecuroides (TASA) on dextran sulfate sodium (DSS)-induced colitis in mice., Methods: Chronic experimental colitis was induced by administration of 4 cycles of 4% DSS. Fifty mice were randomly distributed into 4 groups (normal, DSS, DSS/high-dose TASA, and DSS/low-dose TASA groups) by a random number table with body weight stratification. Mice in the normal group (n=11) and DSS-induced colitis control group (n=15) received control treatment of 20 mL/kg distilled water; DSS plus TASA high- and low-dose groups (n=12 each) were treated with TASA solution (20 mL/kg) at the doses of 60 mg/kg and 30 mg/kg, respectively. The severity of colitis was assessed on the basis of clinical signs, colon length, and histology scores. Moreover, secretory immunoglobulin A (sIgA) and haptoglobin (HP) were analyzed by enzyme linked immunosorbent assay; intercellular adhesion molecule 1 (ICAM-1) and macrophage-migration inhibitory factor (MIF) gene expressions were analyzed by quantitative reverse transcriptase realtime polymerase chain reaction (qRT-PCR) using SYBA green I; and nuclear factor κ B (NF-κ B) expression and activation and p65 interaction with the promoter of ICAM-1 gene were assessed by Western blotting and chromatin immunoprecipitation assay., Results: TASA administration significantly attenuated the damage and substantially reduced HP elevation and maintained the level of cecum sIgA. TASA inhibited the ICAM-1 gene expression and had no effect on MIF gene expression. Also, TASA was able to reduce phospho-I κ B α (p-I κ B α) protein expression; however, it had no effect on the activation of I κ B kinase α (IKK α) and inhibitor of NF-κ B α (I κ B α). Moreover, TASA inhibited the p65 recruitment to the ICAM-1 gene promoter., Conclusions: TASA had a protective effect on DSS-induced colitis. Such effect may be associated with its inhibition of NF-κ B activation and blockade of NF-κ B-regulated transcription activation of proinflammatory mediator gene.

Objective: To study the anti-tumor effect of total alkaloid of Sophora alopecuroides (TASA) on human colon adenocarcinoma SW480 cells and Balb/c nude mice tumor xenograft., Methods: The effect of TASA on cell proliferation was assessed using MTT assay and the cell apoptosis was detected using Annexin V-FITC apoptosis assay. The nude mouse model bearing transplanted solid tumor SW480 was established. The changes of the volume and weight of the tumor were determined after treatment the mice with TASA. Fluorescence quantitative PCR was used to detect Caspase-3, Caspase-9 and BCL-2 mRNA expressions in the tumor., Results: TASA inhibited the proliferation of SW480 cells in a dose- and time-dependent manner. The apoptotic rate of cells was the best when the concentration of TASA was 0.92 mg/mL at 48 hours. The volume and weight of the tumor xenograft in TASA groups were decreased when compared with those of the control group. The results of RT-PCR showed that TASA activated the pro-apoptotic Caspase-3 and Caspase-9 and lowered expressions of BCL-2., Conclusion: TASA can inhibit the growth of SW480 cells and the growth of transplanted solid tumor of human SW480 cell line, the mechanism of which involves the effect of Caspase-3, Caspase-9 and BCL-2 expression.

Sophoridine (SRI), one of the quinolizidine alkaloids, is a new anticancer drug with noticeable antitumor action and lower toxicity. To our knowledge, there is no report about its effect on colitis. Repeated colitis was induced by administration of four cycles of 4% DSS. The severity of colitis was assessed on the basis of clinical signs, colon length and histology scores. Moreover, cecum secretory immunoglobulin A (sIgA) and plasma haptoglobin (HP) were analyzed by enzyme-linked immunosorbent assay and ICAM-1, and macrophage migration inhibitory factor (MIF) gene expression was analyzed by quantitative reverse transcriptase real-time polymerase chain reaction using SYBR Green I. SRI administration significantly attenuated the damage and caused substantial reduction of the rise in plasma HP, and maintained the level of cecum sIgA. SRI inhibited the ICAM-1 gene expression and had no effect on MIF gene expression. In conclusion, for the first time, the activity of SRI on DSS-induced colitis mice was investigated, which suggests that SRI could be an attractive therapeutic option in the treatment of inflammatory bowel disease.

Previous studies have demonstrated that the total alkaloids of Sophora alopecuroides (TASA), which contains many different ingredients like sophocarpine, matrine, oxymatrine, sophoridine, sophoramine, aloperine and cytosine, were able to protect colon against ulcers caused by 2,4,6-trinitrobenze sulphonic acid (TNBS)/ethanol treated models. In order to elucidate the mechanisms by which TASA exerts its effect of anti-inflammation and immunoregulation on rats with colitis, DAI (disease activity index) and histological grading of colitis were evaluated in the animal model. Moreover, the expression of CD4(+)CD25(+) regulatory T cells (Tregs) and IL-10 in rats with experimental colitis were observed by FCM, ELISA and RT-PCR in this study. Results showed that TASA (15, 30 or 60 mg/kg/day) significantly up-regulated CD4(+)CD25(+)Tregs (P = 0.02, P = 0.02, P = 0.03) and IL-10 levels (ELISA: P = 0.03, P = 0.02, P = 0.00; RT-PCR: P = 0.04, P = 0.02, P = 0.01) respectively and decreased the DAI and histological grading of colitis in the peripheral blood (PB) and colon of rat colitis models (3.44 +/- 1.53, 4.25 +/- 1.27, 4.42 +/- 1.24 and 3.50 +/- 1.42, 4.05 +/- 1.32, 4.51 +/- 1.55 vs. 7.18 +/- 1.32 and 7.38 +/- 1.52, P < 0.05, P < 0.01, respectively). Most interestingly, a negative correlation was demonstrated between the expression of CD4(+)CD25(+) Tregs and DAI (Pearson r(PB) = -0.677, P < 0.01; Pearson r(COLON) = -0.663, P < 0.01, n = 60), or histological grading of colitis (Pearson r(PB) = -0.725, P < 0.01; Pearson r(COLON) = -0.623, P < 0.01, n = 60). Simultaneously, a positive correlation existed between CD4(+)CD25(+) Tregs and IL-10 cytokine (IL-10 mRNA) in the colon and PB of rats (Pearson r(PB) = 0.789, P < 0.01, n = 60; Pearson r(COLON) = 0.678, P < 0.01, n = 60). These results may explain to some extent the mechanisms of TASA on treating rats with experimental colitis.

Objective: To prepare colon-targetting tablets of total alkaloids of Sophora alopecuroides and evaluate the effect of pectins of different degree of esterification (DE) on sophoridine release profiles in-vitro., Method: Wet granulation technique was employed to prepare petin-based matrix tablets, then tablets were coated the optimal formulation with Kollicoat MAE 30 DP based on the optimal formulation and analysed their release., Result: Coated formulation E could target total alkaloids of S. alopecuroides to colon and various DE of pectin exerted different effects on sophoridine release. The release of low DE pectin-based matrix tablets coating with Kollicoat MAE 30 DP approximatedly fitted zere-order eqution, which was erosion depended., Conclusion: Low DE pectin-based matrix tablet coating with Kollicoat MAE 30 DP can deliver sophoridine to colon, hence improve the effectiveness of sophoridine.

Objective: To test 5 kinds of alkaloid of Sophora alopecuraides L., the effects of TBSA, MT, OMT, SC, SRI against adenocarcinoma of colon cell line SW620., Methods: SW620 cell line was treated with alkaloid at various concentrations. The effect on cell proliferative activity was measured by MTT assay and the effects of alkaloids against cell growth were compared. The changes of cell morphology were observed through optical microscope and fluorescence microscope and the cell apoptosis was detected by flow cytometry and Annexin V-FITC apoptosis assay after the treatments., Results: According to the results of MTT assay, all of the 5 kinds of alkaloid especially SRI and MT were identified to produce growth inhibition against SW620 cells. The sequence of the intensity of effect was TBSA < OMT < SC < MT < SRI. Further study of SRI and MT demonstrated their effects on inducing SW620 cell apoptosis, and SRI showed the strongest effect. Both SRI and MT induced SW620 cells to apoptosis in a dose-and-time-dependent manner., Conclusion: The 5 kinds of alkaloid especially SRI and MT may inhibit the growth of SW620 cells,and induce SW620 cell apoptosis. The results of this study pave the way for further identification of the effective components in Sophora alopecuraides L. that inhibit colorectal carcinoma both in vivo and in vitro.

Objective: To investigate the effects of matrine on the cell cycle and apoptosis in human colon adenocarcinoma SW620 cells and explore the possible mechanisms., Methods: The effect of matrine on cell proliferation was assessed using MTT assay, and the cell cycle arrest induced by matrine was determined by flow cytometry. The changes of cell morphology were observed through optical microscope, fluorescence microscope and electron microscope, and the cell apoptosis was detected using Annexin V-FITC apoptosis assay., Results: Matrine inhibited the proliferation of SW620 cells in a dose- and time-dependent manner. Compared with the control group, the matrine-treated cells showed increased cell percentage arrested in G 0/G1 phase with decreased S-phase cells. Morphologically, the SW620 cells treated with matrine exhibited cell shrinkage, cell size reduction, plasma condensation, cytoplasmic vacuolar changes, and formation of apoptotic body with also the presence of the signet-ring cells, all typical of apoptotic cells., Conclusion: Matrine exposure of SW620 cells inhibits the cell proliferation, causes cell cycle arrest at G 0/G1 phase, and induces apoptosis in a dose- and time- dependent manner.

Objective: To investigate the effects of total base of Sophora alopecuroides on expression of SOD, MDA, NO, MPO in rats with experimental colitis (UC)., Method: SD rats were randomized into 6 groups and colitis was induced by administrating 2,4,6-trinitrobenzesulphonic acid (TNB) recatlly in rats. Rats were given drug by stomach after the first day for 3 weeks, and sacrificed at 23rd day to determine the content of MDA and activities of SOD, NO, MPO in colonic mucosa, and to estimate the symptom and colonic histology., Result: In UC rats, SOD was significantly decreased whereas MDA, NO, MPO were increased when compared with the normal group (P < 0.05). In all treatment groups, Sophora alopecuroides could increase SOD, decrease MDA, NO, MPO (P < 0.05), and improve the symptoms and histological damages., Conclusion: The total dose of S. alopecuroides may ameliorates or alleviate the symptoms of UC through inhibiting oxygen free radical reaction, decreasing the generation of NO and exerting anti-inflammatory effects.