补肾复方含药血清对大鼠成骨细胞OPG、ODF表达的影响

INFLUENCE OF KIDNEYS-REINFORCING-COMPOSITE-RECIPES-CONTAINING SERUM ON THE EXPRESSION OF RAT OPG AND ODF

为研究补肾复方对大鼠成骨细胞OPG、ODF表达影响。以补肾活血方、补肾健脾方、龟鹿二仙膏、金匮肾气丸、六味地黄丸、左归丸6种补肾中药复方及生理盐水给体重为250g左右的大鼠灌胃,提取含药血清;采用酶消化法从初生2天SD大鼠头盖骨中分离出成骨细胞,用含药血清及生理盐水进行成骨细胞传代培养;取第2代细胞,培养至细胞铺满培养瓶约80%时,更换含药血清培养液24小时;用含药血清培养液24小时后终止培养,提取总RNA,应用RT-PCR方法观察六种补肾复方对大鼠成骨细胞OPG、ODF表达影响。结果显示补肾活血方、补肾健脾方、金匮肾气丸、六味地黄丸和左归丸含药血清培养的大鼠成骨细胞OPG的表达量高于生理盐水对照组,龟鹿二仙膏含药血清培养的大鼠成骨细胞OPG的表达量低于生理盐水对照组,其中补肾活血方组、补肾健脾方组大鼠成骨细胞OPG表达量明显高于生理盐水对照组,统计学有显著性差异(P<0.05)。6种补肾复方含药血清培养的大鼠成骨细胞ODF表达量均低于生理盐水对照组,其中补肾活血方组、补肾健脾方组和金匮肾气丸组大鼠成骨细胞ODF表达量明显低于生理盐水对照组,统计学有显著性差异(P<0.05)。表明补肾活血方、补肾健脾方含药血清能增加大鼠成骨细胞OPG表达量,降低大鼠成骨细胞ODF表达量。

In order to study the influences of composite kidneys-reinforcing recipes on the expression of rat OPG and ODF,70 SD rats were divided into 7 groups: Groups A,B,C,D,E and F taken as the tested and transgastrically given 'BUSHEN HUOXUE FANG','BUSHEN JIANPI FANG','GUILUERXIAN GAO','JINGUISHENQI WAN','LIUWEI DIHUANG WAN' and 'ZUOGUI WAN',respectively and Group G taken as the control and transgastrically given the normal saline.Then the TCD-containing serums were extracted;osteoblasts were separated from the skull of two-day-old SD rat by enzyme digestion;the tested and the control osteoblasts were generatively cultured in the TCD-containing serums and in the normal saline,respectively;when the second-generation cultured cells spread to 80 % of the culture bottle,the TCD-containing serums were changed and the culture was continued for 24 hours and then stopped,the total RNAs were extracted and the influences of the six kidneys-reinforcing recipes on the expression of OPG and ODF were observed by the RTPCR method.The results showed that the OPG expressions of Groups A,B,D,E and F were higher than that of the control,especially those of Groups A and B with a significant statistical difference from the control(P < 0.05),but the one of Group C was lower than that of the control;the ODF expressions of the six tested groups were lower than that of the control,especially those of Groups A,B and D with a significant statistical difference(P < 0.05),suggesting that the TCD-containing serums of 'BUSHENHUOXUEFANG' and 'BUSHENJIANPIFANG' function to increase the rat OPG expression and decrease the rat ODF expression.【