Farrerol inhibits AngII-induced VSMCs proliferation by reducing Cx43 expression / 中国药理学通报

Aim To study the role of Cx43 in inhibi-tion of AngII-induced vascular smooth muscle cells(VSMCs) proliferation by farrerol. Methods The primary VSMCs were isolated and cultured by direct adherent culture methods. VSMCs were identified by immunohistochemstry. The cells were divided into the following groups:control group,AngII group,AngII+Farrerol group. The cell viability was measured by CCK-8 cell vitality test. The proliferation of VSMCs was measured by the methods of Edu. The cell cycle of VSMCs was detected by flow cytometry. The mRNA levels of Cx43 were measured by Real-time PCR. The protein levels of Cx43 were measured by Western blot. Results 60 μmol·L-1farrerol could significantly de-crease the cell viability and EdU rate of VSMCs in-duced by AngII(P<0.05),which could also prevent the transformation of VSMCs from G0/G1phase to S phase. The results of real-time PCR and Western blot showed that,compared with the model group,Farrerol could significantly reduce the mRNA and protein ex-pression level of Cx43(P <0.01). After the interfer-ence of Cx43 by siRNA, the inhibition of proliferation by farrerol decreased significantly. Conclusion Far-rerol inhibits AngII-induced VSMCs proliferation signif-icantly, which might be associated with reducing the expression of Cx43.

Effects of HIF-1αon epithelial-mesenchymal transition, invasion and migration in colorectal cancer cells / 天津医药

Objective To explore whether hypoxia could promote epithelial-mesenchymal transition (EMT) in various differentiated colorectal cancer cells, and analyse the effect of hypoxia on invasion and migration of colorectal cancer cells. Methods HCT116 (poorly differentiated) and HT-29 (highly differentiated) colorectal adenocarcinoma cells were selected respectively. The morphological changes of two cell lines were observed after 0,10,25,50,100 and 150 mg/L cobalt chloride (CoCl2) treatment for 48 h. The expression of hypoxia-inducible factor-1α(HIF-1α) protein was analysed after 0, 10,25,50,100 and 150 mg/L CoCl2 treatment for 48 h. An optimal concentration of CoCl2 was then selected. Methylthiazolyl tetrazolium (MTT) assay was used to detect the proliferation of two kinds of colorectal cancer cells induced by CoCl 2 at different time points (0, 24, 48, 72 and 96 h), and to select an optimal time. Under the optimal concentration and time conditions, the HCT116 and HT-29 cells were processed by hypoxia (hypoxia group) and normoxia (normoxic group). Transwell invasion assay and Wound healing assay were used to detect cell invasion and migration in two groups. Western blot assay and RT-PCR were used to detect protein and mRNA expression levels of HIF-1α, E-cadherin and Vimentin in two groups. Results Two kinds of cells showed obvious morphological changes after 50 mg/L CoCl2 treatment for 48 h. HIF-1αprotein level first increased and then decreased in two groups of cells with the increased concentration of CoCl 2, and 50 mg/L CoCl2 was the optimal concentration (P<0.05). The cell proliferation showed a tendency to decrease after the increase in both kinds of cells with or without hypoxia for 0-96 h (P<0.05), and 48 h was the optimal time. The transmembrane number and cell migration rate were significantly more in hypoxia group than those of normoxic group (P<0.05). The protein and mRNA levels of HIF-1α and Vimentin were significantly higher in hypoxia group than those of normoxic group in HCT116 and HT-29 cell lines (P<0.05). E-cadherin protein and mRNA levels were significantly lower in hypoxia group than those of normoxic group (P<0.05). Conclusion Hypoxia can promote EMT in different differentiated colorectal cancer cells, and can enhance invasion and migration of two kinds of colorectal cancer cells.

Effects of berberine on the proliferation and differentiation of HL-60 cells / 中国药理学通报

Aim To study effects of berberine on the proliferation a nd differentiation of HL-60 cells. Methods Cell inhibitory effec t was determined by growthcurve method and colony formation.Cell differentiation was analyzed by cytomorphological changes, NBT reduction and cell surface antig en.Cell cycle was analyzed by flow cytometry.Results The growth o f HL-60 cells was significantly inhibited by berberine in a time and dose-depe ndent manner.1,2,4,8 mg?L -1 berberine induced differentiation of HL-60 c ells: Morphological evaluation indicated that the berberine-treated cells exhibited granulation appearance; NBT reduction was significantly increase d; expression of CD11b increased obviously. Flow cytometry analysis showed that berberine arrested HL-60 cells in G 0/G 1 phase, the percent of the cells in S phase decreased.Conclusion Berberine can induce differentiatio n and growth inhibition of HL-60 cells