Effects of different compatibility proportion of active ingredients of Kidney Recipe on osteoblast proliferation and differentiation

Jia Ying-min; Li Rui-yu; Wu Mi-shan; Huo Rui-lou; Li Bin

doi:10.3969/j.issn.2095-4344.2015.24.001

Abstract：BACKGROUND:Animal and cel studies have shown that the Kidney Recipe can prevent and treat osteoporosis and improve bone metabolism, but this recipe is complicated. Recent studies on compound Chinese medicine mainly focused on serum drug metabolism and pharmacokinetics, which has limitations, and the effective ingredient and pharmaceutical material basis are uncertain. OBJECTIVE:In the different concentrations and time, by using different compatibility proportion of active ingredients of Kidney Recipe, osteoblasts from neonatal Sprague-Dawley rats were subjected to culture intervention. The proliferation and differentiation of osteoblasts were determined so as to identify time-effect and dose-effect relationship of Kidney Recipe on osteoblasts and to provide experimental evidences for prevention and treatment of osteoporosis. METHODS:Primary neonatal 24-hour osteoblasts of Sprague-Dawley rats were cultured in vitro. Herbs“tonic”and“cathartic”active chemical components of different proportion were used. The experiment contained three groups:Tonics Medicine (T)>Cathartic Medicine (C) group, T<C Medicine group and control group. T>C group and T<C group were given 10, 20, 30μg/L serum-free medium kidney prescription intervention for osteoblast cultures. Cel proliferation was detected at 24, 48 and 72 hours after culture. Alkaline phosphatase secretion of osteoblasts was measured at 48 and 72 hours. RESULTS AND CONCLUSION:At mass concentration of 10μg/L and 48 hours of culture, a significant promoting effect on the proliferation was found in the T>C group (P<0.05). At mass concentration of 20μg/L, no significant difference was detected in the T>C and T<C groups at various time points, but significant difference was found as compared with the control group. At mass concentration of 30μg/L and 48 hours of culture, a significant promoting effect on the proliferation was visible in the T>C group (P<0.05), and the proliferation became weak at 72 hours. At mass concentration of 10μg/L and 72 hours of culture, alkaline phosphatase secretion could be promoted in the T>C group. At mass concentration of 20μg/L and 24 hours of culture, the promoting effect on alkaline phosphatase secretion was most significant in the T<C group (P<0.05). At mass concentration of 30μg/L and 72 hours of culture, the promoting effect on alkaline phosphatase secretion was optimal in the T>C group (P<0.05). Results confirm that active ingredients of Kidney Recipe compatibility proportion can promote osteoblasts proliferation and differentiation most strongly in the T>C group in the time-effect and the dose-effect relationship.