Analysis of the heavy metal contents' effect on steroidal saponins and the anti-breast cancer activity of Paris polyphylla var. yunnanensis.

Background: P. polyphylla var. yunnanensis, as a near-threatened and ethnic medicine in China, used to be a key ingredient in traditional Chinese medicine in treatment of traumatic injuries, sore throat, snakebites, and convulsions for thousands of years. However, there were no reports on the inverse relationship between the contents of heavy metals and saponins and its anti-breast cancer pharmacological activity in P. polyphylla var. yunnanensis. Methods: The present study aimed to reveal the characteristics of heavy metal contents and saponins and its anti-breast cancer pharmacological activity and their interrelationships in P. polyphylla var. yunnanensis from different production areas. The contents of heavy metal and steroidal saponins in P. polyphylla var. yunnanensis were analyzed by inductively coupled plasma mass spectrometry (ICP-MS) and the high-performance liquid chromatography technique, respectively. The Pearson correlation was used to study the correlation between saponins and heavy metals. 4T1 mouse mammary tumor cells were selected and cultivated for antitumor studies in vitro. Cell Counting Kit-8 (CCK-8) assay, Hoechst staining, and flow cytometry analysis were used for the examination of the proliferation and apoptosis of 4T1 tumor cells. Mouse breast cancer 4T1 cells were subcutaneously injected into BALB/c mice to construct a tumor model to explore the in vivo inhibitory effect on breast cancer. TUNEL assay and immunohistochemistry were used for the examination of the effect of P. polyphylla var. yunnanensis from different origins on cancer cell proliferation and apoptosis induction in 4T1 tumor mice. Results: Heavy metal contents were highly correlated with the content of steroidal saponins. The overall content of 10 metals in the three producing origins was of the order C3 >C2 >C1. The total content of eight steroidal saponins in the extracts of P. polyphylla var. yunnanensis from three different origins was C1 >C2 >C3. The Pearson correlation study showed that in all of the heavy metals, the contents of Cd and Ba were positively correlated with the main steroidal saponins in P. polyphylla var. yunnanensis, while Al, Cr, Cu, Fe, Zn, As, Hg, and Pb showed a negative correlation. In vitro experiments showed that the extracts of P. polyphylla var. yunnanensis from three origins could inhibit the proliferation and induce cell apoptosis of 4T1 cells in a concentration- and time-dependent manner, especially in the C1 origin. In vivo experiments showed that the extract of P. polyphylla var. yunnanensis from the three origins could inhibit the growth of tumors and induce the apoptosis of tumor cells. In the three origins, C1 origin had the lowest total heavy metal level but the highest total steroidal saponin level. Therefore, it showed a better effect in reducing the expression of the human epidermal growth factor receptor 2 (HER2) and Kiel 67 (Ki67) and increasing the expression of p53 in tumor tissues compared to the other origins. In conclusion, in the three origins, C1 origin exhibits antitumor pharmacological effects in vivo and in vitro which are better than those in the other origins. Conclusion: In this study, we found that with the increase of the heavy metal content, the content of steroid saponins and anti-breast cancer activity decreased. The results showed that the high content of the total heavy metals may not be conducive to the accumulation of steroidal saponins in P. polyphylla var. yunnanensis and lead to the low anti-breast cancer activity. The results of this study suggest that the content of heavy metals should be controlled in the artificial cultivation process of P. polyphylla var. yunnanensis.

The Effect of Salt-Processed Psoralea corylifolia on Generative Organ Targeting.

Psoralen and isopsoralen are two isomers and main effective components within Psoralea corylifolia. In order to investigate the salt-processing effect on tissue distribution characters of psoralen and isopsoralen, a sensitive and accurate ultrahigh pressure liquid chromatographic tandem mass spectrometric (UHPLC-MS/MS) method has been developed and validated for simultaneous determination of the 2 components in rats' tissues after administration of the extracts that came from either crude or salt-processed Psoralea corylifolia L. Data displayed that both areas under the curve (AUC) of psoralen and isopsoralen from salt-processed scurfpea fruit group were significantly increased compared with that of the crude herb group, especially in heart (p < 0.05), ovary, and testes (p < 0.001). Though the RE and RCmax of psoralen and isopsoralen in all of the investigated organs were over 1.0, generative organs kept the maximum value. The experiment manifested that salt-processing of scurfpea fruit can increase the distribution of psoralen and isopsoralen to generative organs, heart and spleen, and the distribution of psoralen and isopsoralen to generative organs is significantly higher compared to heart and spleen (p < 0.01). Results indicate that salt-processing of scurfpea fruit can significantly increase the distribution of psoralen and isopsoralen to generative organs.

Evaluation of the influence of salt processing on pharmacokinetics of psoralen and isopsoralen in Psoralea corylifolia L.

A sensitive, specific and rapid ultra-high-pressure liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method has been developed to investigate pharmacokinetic properties of psoralen and isopsoralen, two compounds isolated from raw/salt-processed fruit of Psoralea corylifolia L. UHPLC-MS/MS was used with positive ion electrospray. The mobile phase was composed of acetonitrile and 0.1% formic acid aqueous solution and a gradient elution program at flow rate of 0.3 mL/min was applied. Multiple reaction monitoring mode was used for the quantification of psoralen, isopsoralen ([M + H](+) m/z 187.0 → m/z 131.0) and scoparone (m/z 207.0 → m/z 151.1). Scoparone served as an internal standard. The method was fully validated for its sensitivity, selectivity, stability, matrix effect and extraction recovery. The obtained results showed that salt-processed Buguzhi significantly promoted the absorption of psoralen and isopsoralen, and increased the bioavailability of these compounds.

[Effects of sowing date and planting density on the grain' s protein component and quality of strong and medium gluten winter wheat cultivars].

In a field experiment with split-split plot design, the effects of sowing date and planting density on the grain's protein component and quality of strong gluten wheat cultivar Linyou 145 and medium gluten wheat cultivar Linyou 2018 were studied. The results showed that proper sowing date brought the highest protein content and yield in wheat grain. With sowing date postponed, the grain's gliadin and glutenin contents of Linyou 145 increased obviously, while those of Linyou 2018 changed little. The grain quality of Linyou 145 was more affected by sowing date, compared with that of Linyou 2018. When sowing at proper date, the grain's protein and glutenin contents had significant correlations with its wet gluten content, sedimentation value, dough stability time, softness, and evaluation value; while when the sowing date postponed, there existed a positive correlation between the contents of gliadin and wet gluten. The change of the proportions of different protein components in wheat grain induced by the variation of sowing date could be the main reason of the improvement in wheat grain quality. Within the test range (2.25 million - 3.75 million plants x hm(-2)) of planting density, the grain's protein content was less affected, but the grain quality of Linyou 145 was affected to a certain extent. Low planting density (2.25 million plants x hm(-2)) brought the best grain quality of Linyou 2018.