The purpose of this study was to explore the effects of seneciphylline on the proliferation and autophagy of cervical cancer HeLa and Caski cells and the possible mechanisms of autophagy. MTT assay was used to evaluate the effect of seneciphylline on the proliferation of cervical cancer cells. Immunofluorescence assay was applied to investigate the formation of autophagosomes in GFP-LC3/HeLa and GFP-LC3/Caski cells. The effect of seneciphylline on the expression of autophagy-related proteins was checked by Western blotting. In addition, fluorescence colocation assay was used to detect the fusion of autophagosomes and lysosomes. Human cervical cancer subcutaneous xenografts in nude mice were used to evaluate the effect of seneciphylline on the growth of the tumor in vivo. Results showed that HeLa cells proliferation was inhibited by seneciphylline in a dose- and time- dependent manner. Seneciphylline could induce formation of autophagosomes, increase the expression of LC3-II and decrease the expression of P62, suggesting that seneciphylline induced autophagy in HeLa and Caski cells. Compared with seneciphylline alone, seneciphylline combined with later-stage autophagy inhibitor chloroquine significantly increased the expression of LC3-II and P62. Moreover, and fluorescence colocation assay showed that autophagosomes induced by seneciphylline could colocate with lysosomes, indicating that seneciphylline could induce the complete autophagy flux. Compared with seneciphylline alone, seneciphylline combined with earlier-stage autophagy inhibitor 3MA significantly increased the expression of LC3-II and significantly decreased HeLa and Caski cells viability, suggesting that seneciphylline induced protective autophagy. Compared with seneciphylline alone, seneciphylline combined with MEK inhibitor significantly decreased the expression of P-ERK1/2 and formation of autophagosomes, suggesting that autophagy induced by seneciphylline activated MEK/ERK1/2 signal pathway. In addition, seneciphylline showed a significant inhibitory effect on growth of human cervical cancer cells subcutaneous xenografts.

Baihe Dihuangtang is a famous classical formula that has been respected by physicians in the past and is still used today. It was first recorded in ZHANG Zhongjing's Synopsis of the Golden Chamber， and is composed of Lilii Bulbus and Rehmanniae Radix juice. This paper systematically reviewed the research progress of historical evolution， pharmacological activities and clinical applications of Baihe Dihuangtang in recent years， and found that there was no major changes in the composition， decoction method and indications of this formula since the Han dynasty. According to the herbal textual research， the fleshy scaly leaves of Lilium brownii var. viridulum should be selected for Lilii Bulbus in this formula， and the tuberous roots of Rehmannia glutinosa were selected for Rehmanniae Radix. According to the dosage conversion of ancient and modern times， the dosage is 245 g of fresh Lilii Bulbus and 400 g of fresh Rehmanniae Radix， and the ratio of their juice is 1∶1. Its efficacy is to nourish Yin and clear heat， and to tonify the heart and lungs， which is used to treat the heart and lung Yin deficiency syndrome of lily disease. Modern pharmacological studies have shown that the research on the pharmacological effects of Baihe Dihuangtang mainly focuses on anti-depressant， anti-anxiety， improving insomnia and regulating metabolism， and it is mostly used clinically for neurological disorders such as depression， anxiety and insomnia. The quality marker（Q-Marker） of Baihe Dihuangtang were predicted according to the principles of ingredient specificity， component validity， component measurability， formula compatibility， and quality transmissibility and traceability， and it was determined that catalpol， rhmannioside D， regaloside A， regaloside B， regaloside C， and acteoside could be selected as potential Q-Markers of Baihe Dihuangtang， which could provide scientific reference for the establishment of the quality control system and the development of compound preparation of this famous classical formula.