[Inhibitory effect of paeoniflorin on CAL27 cells of tongue squamous cell carcinoma and its mechanism].

PURPOSE: To investigate the inhibitory effect of paeoniflorin(PF) with different concentrations on CAL27 cells of tongue squamous cell carcinoma in vitro and its possible mechanism. METHODS: CCK-8 technique and clone formation trial were used to detect the effect of PF on the proliferation and clone formation of CAL27 cells. Scratch test and Transwell method was used to detect the effects of PF on migration and invasion of CAL27 cells. Staining of Hoechst33342 was employed to evaluate the influence of PF on apoptosis of CAL27 cells, while Western blot was utilized to investigate the effect of PF on the expression of NF-κB pathway related proteins and EMT related proteins. The effect of PF on NO production in CAL27 cells was detected by nitric oxide detection kit. Statistical analysis was performed with SPSS 27.0 software package. RESULTS: PF inhibited proliferation, migration, and invasion of CAL27 cells in vitro in a concentration-dependent way. Moreover, PF caused apoptosis of CAL27 cells. PF impeded NF-κB pathway activity, decreased the expression of P-P65, further reduced the expression of iNOS and MMP-9, suppressed the production of NO, and concurrently inhibited Vimentin,promoted E-cadherin. CONCLUSIONS: Paeoniflorin inhibits the proliferation, migration and invasion of CAL27 cells, which may play an anti-cancer role by inhibiting the activation of NF-κB pathway and EMT.

[The effect of osthole on lipopolysaccharide-induced macrophages polarization and inflammatory reaction].

PURPOSE: To investigate the effect of Osthole (OST) on lipopolysaccharide (LPS)-induced macrophage polarization and inflammatory reaction. METHODS: The effect of different concentrations of OST on proliferative activity of RAW264.7 macrophages was examined by CCK-8 method; the effect of OST at different concentrations (6.25, 12.5 and 25 µmol/L) on macrophage polarization and inflammation was investigated by using intracellular reactive oxygen species (ROS) detection kit (DCFH-DA), immunofluorescence staining, q-PCR and flow cytometry. The effects of OST on macrophage polarization and inflammatory responses were investigated by immunoblotting of proteins. Graphpad prism 8.0 software package was used for statistical analysis of the data. RESULTS: CCK-8 results showed that OST was not significantly cytotoxic to RAW264.7 at less than 25 µmol/L, immunofluorescence and q-PCR results showed that OST at 6.25, 12.5 and 25 µmol/L inhibited the expression of inflammatory factors in M1 macrophages, and iNOS, TNF-α, CCR7 were reduced in a concentration-dependent manner, and effectively upregulated the expression of M2 inflammatory factors IL-10, Arg-1 and CD206. Flow cytometry showed that OST effectively inhibited the expression of LPS-induced M1 marker CD86 in macrophages. CONCLUSIONS: OST can regulate lipopolysaccharide-induced M1 macrophages polarization and reduce inflammatory reaction.

[Experiment of osteogenic differentiation of human adipose stem cells in vitro].

PURPOSE: To explore the feasibility of isolation, proliferation and osteogenic potential of human adipose stem cells (hASCs) in vitro. METHODS: hASCs were isolated from the adipose tissue of liposuction aspirated by means of enzymatic digestion and proliferated in vitro. Morphology and growth kinetics by hemacytometer of primary and continuous cells were observed. Osteogenic differentiation of hASCs cultured in conditional culture medium was evaluated by Alkaline phosphatase (ALP) staining and Von Kossa for mineralization, immunohistochemistry of osteocalcin (OCN), ostseopotin (OPN) and collagen type I for matrix maturation, which was further confirmed by RT-PCR analysis for markers of osteogenic differentiation, with the non-osteogenic induced cells as controls. RESUITS: It was shown that both primary and passage cells shared similar fibroblastic-like morphology, while passage cells grew faster. In osteogenic differentiation bioassay, there was a significant difference in expression of ALP, OPN, and OCN between groups when the culture time was extended to 14 days, and collagen type I was expressed in both groups. CONCLUSIONS: These results indicate that the hASCs grow well in vitro and could be induced to osteoblastic differentiate. hASCs can serve as a seeding cell source for bone engineering.