Protection of icariin on impairment of rat bone mesenchymal stem cells induced by 5-fluorouracil in vitro / 中国药理学通报

ABSTRACT

Aim To study the protective effect of icariin on the impairment of rat bone mesenchymal stem cells (BMSCs) induced by 5-fluorouracil (5-FU) and the underlying mechanism. Methods The proliferation of BMSCs was evaluated by MTT assay, the morphological change of apoptotic cells was tested by using the chromatin dye Hoechst 33258, and the apoptotic rate was analyzed by flow cytometry. The proteins expression of cleaved caspase-3 was detected with Western blotting, the activity of caspase-3 was detected by Activity Assay Kit; iNOS and TNOS were detected by nitric oxide synthase (NOS) assay kit. Results MTT detection showed that 5-FU could reduce the survival rate of BMSCs, 10, 20, 40 μmol · L-1ICA could promote the proliferation of BMSCs, and blocked the inhibition of BMSCs cells growth induced by 5-FU. When BMSCs cells were treated by both ICA (10, 20 p, mol · L-1) and 5-FU (2.5×l0-2g· L-1) for 48 hours, compared with model group, the early apoptotic rates of ICA test groups were significantly reduced (P < 0. 01), the expression of cleaved caspase-3 and the activity of caspase-3 were significantly reduced (P < 0. 01), and the activity of iNOS and TNOS was inhibited in a certain degree (P < 0. 05). Conclusions ICA could protect BMSCs against 5-FU-induced damage. The protective role of ICA is probably related to reducing the activity of iNOS and TNOS, inhibiting the activation of caspase-3, and inhibiting the occurrence of apoptosis.