骨金散对去卵巢大鼠Ⅰ型胶原交联N-端肽、Ⅰ型胶原mRNA表达的影响

The effects of Gujinsan on the level of serum cross linked N-telopeptide of type I Collage and the expression of type I collagen mRNA in bone tissue in ovariectomized rats

目的:复制去卵巢大鼠骨质疏松动物模型,探讨中药骨金散对去卵巢雌性大鼠血清Ⅰ型胶原交联N-端肽(NTX)和骨组织Ⅰ型胶原mRNA表达的影响。方法:6月龄雌性SD大鼠40只,随机分为去势组和假手术组。去势组切除双侧卵巢,假手术组切除部分脂肪。大鼠去势后1个月,将去势组随机分为3组:模型组、雌激素组和骨金散组,雌激素组每日灌服己烯雌酚0.05m g/kg,骨金散组按1.8g/kg剂量灌服骨金散,其余各组灌服等量生理盐水。2个月后,酶联免疫吸附法测量血清碱性磷酸酶(ALP)、NTX水平,RT-PCR方法检测右侧股骨骨组织Ⅰ型胶原mRNA表达水平。结果:与假手术组比较,模型组血清ALP、NTX含量明显升高(P<0.01),Ⅰ型胶原mRNA水平显著下降(P<0.01)。雌激素组和骨金散组大鼠体内的ALP、NTX水平显著低于模型组(P<0.01),骨金散组大鼠体内的ALP水平高于雌激素组(P<0.05)。雌激素组和骨金散组实验大鼠骨组织中Ⅰ型胶原蛋白mRNA表达含量明显高于模型组(P<0.01)。结论:骨金散可减轻去卵巢大鼠ALP、NTX水平的提高,上调骨组织Ⅰ型胶原mRNA的表达水平,这可能与其治疗骨质疏松的机制有关。

Objective：To establish the animal models of osteoporosis in ovariectomized rats and investigate the effects of Gujinsan on the level of serum alkaline phosphatase（ALP）,cross linked N-telopeptide of type I collage（NTX） levels,and the expression of type I collagen mRNA in bone tissue.Methods：40 six-month old SD female rats were randomly assigned to two groups,including the ovariectomized group and false operation group.In the ovariectomized group,double ovaries of rats were removed.In the false operation group,some fat around the ovaries on both sides of rats were removed.At 4 weeks after operation,the ovariectomized groups were randomly allocated into ovariectomized control group,estrogen group,and Gujinsan group,by drug intervention are as follows.Estrogen group：intragastric administrated stilboestrol（0.05 mg/kg per day）.Gujinsan groups：intragastric administrated Gujinsan（1.8g/kg per day）.The rest of groups：intragastric administrated the same amount of normal saline.Two months later,alkaline phosphatase（ALP） and cross linked N-telopeptide of type I collage（NTX） levels were detected by enzyme-linked immunosorbent assay（ELISA）,and the expression of type I collagen mRNA in bone tissue were detected by reverse transcription-polymerase chain reaction（RT-PCR）.Resultss：In ovariectomized control group,the serum ALP,NTX were significantly higher（P0.01）,and the type Ⅰ collagen mRNA levels were significantly lower（P0.01） than that in false operation group.The serum ALP,NTX levels in estrogen group and Gujinsan group of rats were significantly lower than that in ovariectomized control group（P0.01）,and the serum ALP levels in Gujinsan group were higher compared with the estrogen group（P0.05）.The type Ⅰ collagen mRNA expression in bone tissue both in estrogen group and the Gujinsan group rats were significantly higher than that in ovariectomized control group（P0.01）.Conclusion：Gujinsan could reduce the higher levels of serum ALP and NTX and can increase the level of type Ⅰ collagen mRNA expression in ovariectomized rats,which might be one of the mechanisms of Gujinsan for the treatment of osteoporosis.