| Research purpose The current study investigated the effects of Zuogui pill, which is a representive prescription to invigorate kidney and nourish essence, on aging rat’s bone marrow mesenchymal stem cells (MSC) proliferation, Osteogenic differentiation and Adipogenic differentiation, under the guidance of the theories of traditional Chinese medicine(i. e., kidney essence deficiency caused aging)and modern biology (i.e., stem cell has the congenital essence attribute and exists in the form of the cellular level, The study combined traditional Chinese medicine in anti-aging and stem cell research to assess the effects of the invigorating kidney and Nourishing essence on aging MSC activity and functions from the perspective of the cell biology, through analyzing related indicators to assess Zuogui pill anti-aging effect. The current study attempted to provide valid experimental evidence for traditional Chinese medicine clinical prevention and treatment of aging.Methodolog1, The aging animal samples were Sprague-Dawley rats aged3months, half for male, half for female. The samples were intraperitoneally injected with D-galactose. Some of them were fed with the water decoction of Zuogui pill.2, MSC were stained by Giemsa method, and were observed by the Optical microscope and electron microscope respectively.3, We observed control group and experimental group in terms of cellular form, growth curve, growth cycle and-galactosidase staining senescent cells. Based on the these data, we compared the MSC proliferation capacity of normal, model and Zougui groups in the experiments.4, We compared differentiation capacity of normal group, model group, and Zuogui Pill group by examing the expression of the ratio of ALP stained positive cells and alkaline phosphatase(ALP) activity. 5, We also compared differentiation capacity into steatoblast of normal, model and Zougui group by examining the ratio of oil red stained positive cells and the expression of triglyeride(TG).Findings1, Under the Optical microscope, MSC of model group presented morphological characteristics of senescent cells.While those in Zuogui pill groups were closed to the normal group. Under the electron microscope, MSC of model group presented senescent cells morphological characteristics, But the ultrastructure of the Zuogui pill groups BMSC were closed to the normal group.2, The growth curve of P3and P5MSC shows that, the proliferation capacity of MSC of model group was significantly lower than those of the normal group and Zuogui pill group (P<0.05). The results ofβ-Galactosidase staining showed that model group of aging cells has the highest rate, the Zuogui pill Significantly reduce the positive rate of senescent cells.(P<0.05).3, The basic carbonate enzyme of MSC of different groups was positive after induction. On7d of differentiating into osteoblasts, the rate of MSC ALP expression in the model group was obviously Lower than the normal group, and Zuogui pill group (P<0.05)。 On14d of osteoblasts induction, the rate of ALP stained positive cells of MSC in the mode1group was obviously lower than the normal group and Zuogui Pill group (.P<0.05).4, The oil red stained positive cells of different groups of rats were negative after induction. On7d of differentiating into osteoblasts, MSC TG expression in the model group was obviously higher than the normal group, and Zuogui pill group (P<0.05)。 On the7d of adipogenic differentiation, the rate of oil red stained positive cells in model group MSC was obviously higher than the normal group and Zuogui pill group (P<0.05)。Conclusions1, The water solution of Zuogui pill could delay the aging of rat MSC.2, Aging rat MSC proliferation ability lessened. The water decoction of Zuogui pill could improve the MSC proliferation ability of aging rats to some extent.3, Compared with MSC of normal rats, the Model group’s ability of differentiation into adipoeytes of MSC decreased, while the ability of differentiation into Osteoblast of MSC from aging rats increased The water decoction of Zuogui pill with certain level could facilitate osteogenic differentiation of MSC in aging rats and inhibit its adipogenic differentiation. |
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