BACKGROUND:Immortalized cervical epithelial cels H8 can become cancerous under the induction of carcinogenic agent, and may cause cervical cancer when there is a cofactor interaction. However, there is stil a lack of effective intervention for female patients with precancerous lesions, and this treatment is blank in the clinic. OBJECTIVE: To explore the effects and mechanism of astragalus injection on apoptosis of human immortalizedcervical epithelial cels H8. METHODS: This study contained two groups: astragalus drug group and the blank control group. (1) Enzyme linked immunosorbent assay (ELISA) was used to detect DNA fragments of apoptotic H8 after astragalus injection. (2) Enzyme-labeling instrument was used to analyze the changes in caspase-3 and caspase-9 activities a fter astragalus injection. (3) Western blot assay was used to detect the protein expression changes of caspase-3, caspase-9 and PARP in H8 cels after astragalus injection. RESULTS AND CONCLUSION: (1) ELISA results showed that at 0, 6, 12 and 24 hours after 20 g/L astragalus injection, DNA fragments were gradualy increased with time prolonged in a time-dependent effect (P < 0.05). (2) Enzyme-labeling instrument demonstrated that at 0, 6, 12 and 24 hours after 20 g/L astragalus injection, caspase-3 and caspase-9 activities increased in a time-dependent manner (P < 0.05). (3) At 0, 6, 12 and 24 hours after 20 g/L astragalus injection, the expression of cleaved caspase-3 and cleaved caspase-9 were gradualy increased in H8 cels (P < 0.05). Cleaved PARP protein expression was gradualy decreased (P < 0.05). These findings indicate that astragalus injection could obviously induce H8 apoptosis, which may be associated with the upregulated protein expression of caspase-3 and caspase-9.

In vitro experiments are an indispensable method in the pharmacological research of Chinese materia medica. However， the development of in vitro experiments is hindered by the various sources， complex composition， and unstable physical and chemical properties of Chinese materia medica. Therefore， it has become an urgent problem to develop reasonable methods for in vitro pharmacological research. At present， the commonly used in vitro experimental methods in the pharmacological research of Chinese materia medica include direct administration and indirect administration. Indirect administration methods include drug-containing serum/plasma， drug-containing intestinal absorption solution， and drug-containing liver incubation solution. With the development of computer technology and systems biology， the combination of high performance liquid chromatography-mass spectrometry （HPLC-MS）， omics， network pharmacology， computer/artificial intelligence-assisted drug design and other technologies with in vitro experiments of Chinese materia medica has not only greatly reduced the research cost but also significantly improved the efficiency of pharmacological exploration， effective ingredient screening， and drug discovery. Therefore， the combined methods have been gradually recognized and widely used. This paper reviewed the in vitro experimental methods in pharmacological research of Chinese materia medica and their progress， aiming to provide help for selecting reasonable research methods.