CAJ | 학술논문

目的研究含壮骨强筋片药物血清对老年大鼠骨髓基质干细胞(BMSCs)增殖及成骨分化的影响.方法取18月龄SD大鼠培养BMSCs,流式细胞仪检测细胞表面蛋白标志CD34、CD31、CD29；CCK-8法检测不同浓度(2.5％、5％、10％)含药血清于培养第24、48、72 h细胞的增殖情况,分为成骨诱导组、成骨诱导中药组、中药组、空白组.培养7d后,检测碱性磷酸酶(ALP)活力.14 d后,检测ALP和骨钙素(Osteocalcin,OC) mRNA含量.结果 CD29表达为阳性,CD31和CD34表达均为阴性.2.5％、5％的含药血清均可促进细胞增殖.ALP活力检测(U/gprot):成骨诱导组(202.76±15.44)、成骨诱导中药组(240.48±18.55)、中药组(178.87±17.29]均明显高于空白组(111.24±20.71)(t=22.50、7.985、3.535,均P＜0.01)；成骨诱导中药组高于成骨诱导组(t=3.103,P＜0.05).ALP基因相对吸光度值:成骨诱导组(0.40±0.20)、成骨诱导中药组(0.60±0.06)均高于空白组(0.09±0.03)(t=2.372、9.547,均P＜0.01)；成骨诱导中药组明显高于成骨诱导组(t =2.523,P＜0.05).OC基因相对吸光度值:成骨诱导组(0.58±0.09)、成骨诱导中药组(0.76±0.11)均高于空白组(0.41±0.02) (P ＜0.05,P＜0.01),成骨诱导中药组明显高于成骨诱导组(t=2.673,P＜0.05).结论含壮骨强筋片药物血清可促进老年大鼠骨髓基质干细胞的增殖及成骨分化,可能是其防治老年性骨质疏松症的机制.
목적 연구함장골강근편약물혈청대노년대서골수기질간세포(BMSCs)증식급성골분화적영향.방법 취18월령SD대서배양BMSCs,류식세포의검측세포표면단백표지CD34、CD31、CD29；CCK-8법검측불동농도(2.5％、5％、10％)함약혈청우배양제24、48、72 h세포적증식정황,분위성골유도조、성골유도중약조、중약조、공백조.배양7d후,검측감성린산매(ALP)활력.14 d후,검측ALP화골개소(Osteocalcin,OC) mRNA함량.결과 CD29표체위양성,CD31화CD34표체균위음성.2.5％、5％적함약혈청균가촉진세포증식.ALP활력검측(U/gprot):성골유도조(202.76±15.44)、성골유도중약조(240.48±18.55)、중약조(178.87±17.29]균명현고우공백조(111.24±20.71)(t=22.50、7.985、3.535,균P＜0.01)；성골유도중약조고우성골유도조(t=3.103,P＜0.05).ALP기인상대흡광도치:성골유도조(0.40±0.20)、성골유도중약조(0.60±0.06)균고우공백조(0.09±0.03)(t=2.372、9.547,균P＜0.01)；성골유도중약조명현고우성골유도조(t =2.523,P＜0.05).OC기인상대흡광도치:성골유도조(0.58±0.09)、성골유도중약조(0.76±0.11)균고우공백조(0.41±0.02) (P ＜0.05,P＜0.01),성골유도중약조명현고우성골유도조(t=2.673,P＜0.05).결론 함장골강근편약물혈청가촉진노년대서골수기질간세포적증식급성골분화,가능시기방치노년성골질소송증적궤제.
Objective To study the effects of serum of the Zhuartgguqiang jin tablets on the proliferation and osteogenic differentiation of bone marrow stromal stem cells (BMSCs) of aged rats.Methods The BMSCs were obtained from male SD rats of 18 months.The cell surface markers CD34,CD31,CD29 were detected by flow cytometry.The proliferation of BMSCs treated by different concentration of medicated serum (2.5％,5％,10％) at 24,48,72hours was detected by CCK-8 method.The BMSCs were divided into 4 groups,which were osteogenic induction group,the Chinese medicated osteogenic induction group,Chinese medicine group,the blank group.After the BMSCs were cultured for 7 days,alkaline phosphatase (ALP) vitality was detected.After the BMSCs were cultured for 14 days,the BMSCs were stained by alizarin red,the mRNA of ALP and osteocalcin(OC) was detected.Results The expressionof CD29 was positive,and the expression of CD31 and CD34 were negative.2.5％,5％ of the medicated serum could promote cell proliferation.The ALP vitality of osteogenic induction group[202.76 ± 15.44(U/gprot)],the Chinese medicated osteogenic induction group [240.48 ± 18.55 (U/gprot)],Chinese medicine group [178.87 ± 17.29 (U/gprot)]were significantly higher than that of blank group [111.24 ± 20.71 (U/gprot)] (t =22.50,7.985,3.535,all P ＜0.01).The ALP activity of Chinese medicated osteogenic induction group was higher than that of osteogenic induction group (t =3.103,P ＜ 0.05).The ALP gene relative OD value of osteogenic induction group (0.40 ± 0.20) and Chinese medicated osteogenic induction group (0.60 ± 0.06) were higher than the blank group (0.09 ± 0.03) (t =2.372,9.547,all P ＜0.01).The ALP gene expression of Chinese medicated osteogenic induction group was obviously higher than that of the osteogenic induction group(P ＜0.05).The OC gene relative OD value of osteogenic induction group(0.58 ± 0.09) and Chinese medicated osteogenic induction group(0.76 ± 0.11) were higher than the blank group(0.41 ± 0.02) (P ＜ 0.05,P ＜ 0.01).The OC gene expression of Chinese medicated osteogenic inductiongroup was obviously higher than that of the osteogenic induction group(t =2.673,P ＜ 0.05).Conclusion Medicated serum of Zhuangguqiangjin tablets can promote the proliferation and osteogenic differentiation of BMSCs of aged rats,which may be the mechanism of prevention senile osteoporosis.