菟丝子含药血清对大鼠胚胎肢芽形态发育的影响

目的探讨不同剂量菟丝子含药血清对SD大鼠胚胎肢芽形态发育的影响。方法采用中药血清药理学方法制备菟丝子含药血清。取孕第15天胚胎前肢芽,随机分为3个实验组、阴性对照组和阳性对照组。实验组和阴性对照组分别加入10%培养体积的高、中、低剂量菟丝子含药血清和正常血清,阳性对照组加入全反式维甲酸溶液(10 mg/mL)。采用体外肢芽悬浮培养法培养72h后收获肢芽,石蜡包埋、切片、HE染色观察肢芽细胞和软骨形态。每组取30个肢芽,测量肢芽长度。结果经HE染色后,菟丝子含药血清低、中剂量组和阴性对照组肢芽结构完整,细胞排列整齐,软骨轮廓清晰,指突明显。高剂量组和阳性对照组肢芽软骨出现,轮廓较清晰,但指突不明显。分段测量肢芽长度后,与阴性对照组比较,低和中剂量组前肢芽近端a段长度和远端b段长度无显著性差异(P>0.05)。而阳性对照组和高剂量组的肢芽长度显著小于阴性对照组,与之比较差异显著(P<0.05)。结论高剂量菟丝子含药血清对SD大鼠胚胎前肢芽生长有抑制作用,可显著抑制肢芽指突的生长,存在潜在的胚胎发育毒性。

Effect of cuscuta chinensis Lam medicated serum on morphology development of rat embryo limb buds

Objective:To explore the effect of Cuscuta Chinensis Lam medicated serum on the morphology development of SD rat embryo limb buds.Methods:The different doses Cuscuta Chinensis Lam medicated serum were prepared by serological pharmacological method.The forelimb buds were amputated from SD rat embryos on day15.Five test groups included the experimental groups which were cultured separately by low,middle,high doses Cuscuta Chinensis Lam medicated serum,negative control group which was cultured by normal serum and all-trans retinoic acid(10 mg/mL)was chosen as positive standard control.The forelimb buds were cultured in vitro for 72 hours,limb buds of each group stained with HE to observe development of limb buds;30 limb buds were measured the length.Results:Staining with HE,long bones of each limb buds can be observed,but no carpal.Cells of the low,middle and negative control group were arranged in rows.The cartilage shapes were clear and obvious.However,the cartilage shape of high dose group were not so clear,the fingers were not obvious.By measuring the length of forelimb buds,compared to negative control group,the limb buds length of low and middle groups had no significantly difference(P>0.05.However,the length of positive and high groups were significantly shorter than negative control group(P<0.05).Conclusion:High dose of Cuscuta Chinensis Lam medicated serum can inhibit the development of rat embryos forelimb buds length,especially curb the finger development,implying that it has potential embryo developmental toxicity.