左归丸含药血清对高糖负荷ICR小鼠早期胚胎发育的影响

Effects of serum containing Zuogui Pills on development of ICR mouse embryos in high glucose

目的:研究左归丸含药血清对高糖负荷下ICR小鼠早期胚胎发育的影响。方法:使用含0、20、40、60mmol/L葡萄糖的人类输卵管液培养液(HTF)对ICR小鼠胚胎细胞进行体外培养,观察2-细胞率、囊胚细胞率、囊胚细胞数的变化。选择显著抑制ICR小鼠早期胚胎发育的适宜葡萄糖浓度同时添加5%或10%左归丸含药血清体外作用于胚胎1-细胞,观察2-细胞率、囊胚细胞率、囊胚细胞数的变化。结果:胚胎细胞在添加0、20mmol/L葡萄糖的HTF培养基中可正常孵化、发育,在含40、60mmol/L葡萄糖的HTF培养基中发育受到明显损伤(P<0.05),左归丸含药血清能够降低含40及60mmol/L葡萄糖的HIF培养基对胚胎发育的损伤(P<0.05)。结论:左归丸含药血清对高糖诱导的胚胎细胞发育损伤具有明显保护作用,在分子水平揭示了中医滋阴补肾方降糖作用的科学内涵。

Objective: To study the effect of serum containing Zuogui Pills on development of ICR mouse embryos cultured in high glucose medium in vitro. Methods: One-cell embryos were collected and cultured in HTF supplemented with 0, 20, 40, 60mmol/L glucose respectively. The culture efficiency was evaluated by determining the proportion of embryos reaching the tow-cell, blastocyst. The total cell numbers were counted in the blastocystembryos. The glucose concentration that can markedly inhibit the early embryonic development of ICR mouse were selected. The changes of cleavage rate, blastocyst rate and number of blastodermal cells were observed after serum containing 5% or 10% Zuogui Pills treatment. Results: Addition of glucose to HTF in a concentration as high as 20mmol/L didn't inhibit the development of ICR mouse one-cell to hatched blastocyst development. HTF supplemented with 0, 20, 40, 60mmol/L glucose respectively had detrimental effects on preimplantation embryo development and serum containing Zuogui Wan protected pre-implantation embryo development from 40, 60mmol/L glucose induced impairment. Conclusion: The high concentration of glucose is toxic to the early embryonic development and serum containing Zuogui Wan Pills inhibit the impairment induced by high concentration of glucose, which provide further insights into mechanism of blood glucose reducing effects of the TCM yin-nourishing and kidney-invigorating prescription at the molecular level.