人参皂苷Rg_1、甲壳胺和转化生长因子β_1对人牙周膜细胞增殖和分化功能的作用

Effects of ginsenoside Rg_(1) , chitosan and TGF-β_(1) on the proliferation and differentiation of human periodontal ligament cells

目的 :探讨人参皂苷Rg1、甲壳胺 (Chi)和转化生长因子 β1(TGF β1)对原代培养的人牙周膜细胞增殖和分化功能的影响。 方法 :用原代培养的人牙周膜细胞 ,采用噻唑蓝 (MTT)比色法、酶动力学方法、放射免疫法和流式细胞仪检测人参皂苷Rg1(0 .0 1μmol/L)、不同浓度Chi(0 .0 5 g/L、0 .1g/L、0 .2 g/L)、不同浓度TGF β1(0 .5 μg/L、1μg/L、2 μg/L)和单纯DMEM培养液 (对照组 )对人牙周膜细胞增殖、碱性磷酸酶 (ALP)活性、骨钙素分泌和细胞凋亡的影响。 结果 :①与对照组比较除TGF β1(0 .5 μg/L、2 μg/L)组和Chi(0 .2 g/L)组第 7天外 ,TGF β1、Chi各浓度组及人参皂苷Rg1组在 3、5、7天均能明显增强牙周膜细胞的增殖能力 (P <0 .0 5 )。②TGF β1、Chi各浓度组及人参皂苷Rg1组细胞裂解液中ALP活性均明显高于对照组 (P <0 .0 5 )。③ 7天时 ,除Chi(0 .2 g/L)组外 ,其他各组骨钙素分泌均明显强于对照组 (P <0 .0 5 ) ;2 1天时 ,人参皂苷Rg1(0 .0 1μmol/L)组明显高于其他各组 (P <0 .0 5 )。④TGF β1(1μg/L)组、Chi(0 .1g/L)组和人参皂苷Rg1(0 .0 1μmol/L)组细胞凋亡百分率与对照组比较均明显降低 (P<0 .0 1)。 结论 :与TGF β1一样 ,Chi及人参皂苷Rg1有促进牙周膜细胞增殖的作用 。

Objective: To observe the effects of ginsenoside Rg_(1) , chitosan and TGF-β_(1) on the proliferation and differentiation of human periodontal ligament cells( PDLC). Methods:Human periodontal ligament cells were isolated and cultured. The effects of ginsenoside Rg_(1)(0.01 μmol/L), chitosan((0.05 g/L,)0.1 g/L, 0.2 g/L) and TGF-β_(1)(0.5 μg/L, 1 μg/L, 2 μg/L) on the proliferative ability of human PDLCs were evaluated with MTT method. The alkaline phosphatase activities of human PDLCs were measured with spectrophotometric assay. The secretion of osteocalcin of human PDLCs were measured with radioimmunological method and the apotosis rates of human PDLCs were assayed with flow cytometry with PI staining method. Results: ①Comparing with the control group, the proliferative ability of human PDLCs in ginsenoside Rg_(1)(0.01 μmol/L)group ,Chi(0.05 g/L, 0.1 g/L, 0.2 g/L) groups and TGF-β_(1 )((0.5 μg/L), 1 μg/L, 2 μg/L) groups on day 3,5,7 were considerably increased (P< 0.05) , except those in TGF-β_(1 )(0.5 μg/L、2 μg/L) and Chi(0.2 g/L) groups on day 7 . ② Alkaline phosphatase activity of human PDLCs in ginsenoside Rg_(1 )group , all chitosan groups and all TGF-β_(1) groups were significantly enhanced (P < 0.05).③ Comparing with the control group, the secretion of osteocalcin of human PDLCs in the ginsenoside Rg_(1 )group , all chitosan groups and all TGF-β_(1) groups except that in Chi((0.2 g/L)) group were obviously promoted(P<0.05)on day 7. On day 21,the secretion of osteocalcin of human PDLCs in ginsenoside Rg_(1)(0.01 μmol/L) was higher than that in the other groups ((P< 0.05)). ④Comparing with the control group, apotosis rates of human PDLCs in ginsenoside Rg_(1)((0.01 μmol/L)) group , Chi( 0.1 g/L) group and TGF-β_(1) (1 μg/L) group were considerably decreased (P < 0.01). Conclusion: These findings suggested that ginsenoside Rg_(1) , chitosan and TGF-β_(1) are capable for producing an accelerative effect on the proliferation and differentiation functions of human PDLCs.