Effect Of Angelica Polysaccharide On Proliferation And Apoptosis Of Osteoblast Cultured In Vitro

Objective:This study researched the effect of different concentration of Angelica Polysaccharide on the proliferation and apoptosis of human osteoblasts in vitro,and then to studied the effects of Angelica polysaccharides on cell cycle and expression of related genes(Cyclin Dl,Bcl-2,Bax)in order to explore the role and possible mechanism of Angelica polysaccharides in the treatment of osteoporosis.Method:(1)Human osteoblast cell lines were cultured in vitro.The cells were randomly divided into control group and experimental group.The experimental groups were cultured in DMEM with different concentration of Angelica polysaccharide(0.25,0.5,1,10 mg/ml)for 72h,MTS method was adopted to detect cell cytotoxicity.(2)Human osteoblast cell lines were cultured in vitro.The cells were randomly divided into control group and APs group(0.5 mg/ml),cultured for five days;MTS was used to detect the proliferation of osteoblasts,flow cytometry was adopted to detect the effect of Angelica polysaccharides on cell cycle and apoptosis,Finally,q-PCR and Western blot were used to detect the effects of Angelica polysaccharides on the expression of Cyclin D,Bcl-2and Bax.Results:(1)MTS assay have showed that Angelica polysaccharide(0.25-lmg/ml)could promote cell proliferation,especially at 0.5 mg/ml concentration.However,a higher dose of Angelica polysaccharide had cytotoxicity and inhibited the proliferation of osteoblasts.(2)Angelica polysaccharide(0.5mg/ml)was used to stimulate osteoblasts for 5 days,MTS assay found that:as time goes on,the effect is more obvious,especially on the fifth day(p<0.01).(3)Human osteoblasts were intervened with Angelica polysaccharide(0.5mg/ml)for 5 days,then the cell cycle distribution were detected by flow cytometry;the results showed that the ratio of S phase cells in the control group was(27.70±0.6658)%,but the proportion of S phase cells in Angelica polysaccharide group was(38.86±0.9557)%,the difference was statistically significant(p<0.05)(4)Human osteoblasts were intervened with Angelica polysaccharide(0.5mg/ml)for 5 days,and the proportion of apoptosis were detected by flow cytometry,the results showed that the ratio of apoptotic cells in control group was(10.49±1.011)%,and the proportion of apoptotic cells in the Angelica polysaccharide group was(6.233±1.621)%,the difference was not statistically significant.(5)Human osteoblasts were intervened with Angelica polysaccharide(0.5mg/ml)for 5 days,the expression of CyclinDl,Bcl-2 and Bax was detected by q-PCR and Western blot.The results showed that:compared with the control group,the expression of CyclinD1 in angelica polysaccharide group was significantly up-regulated(p<0.05),but there was no significant change in the expression of Bcl-2 and Bax(P>0.05).Conclusion:Angelica polysaccharides in a certain concentration range can promote the proliferation of human osteoblasts in vitro,the mechanism of action may be related to the promotion of CyclinD1 expression,which may be related to the distribution of G1 phase to the S phase.It also provides a theoretical basis for the treatment of osteoporosis.