恐伤孕鼠对其仔鼠颅骨培养成骨细胞Cx43、BGP表达的影响

Effects of Horror Injuring Pregnant Rats on Expressions of Cx43 and BGP in Filial Rats Calvarial Osteoblasts

目的:从先天肾虚影响颅骨培养成骨细胞Cx43、BGP表达情况入手,研究其影响骨早期发育的机制,并论证补肾填精法的作用。方法:采用模拟地震振动、光电刺激复合应激恐吓孕鼠造成子代先天肾虚。将孕鼠分为正常、模型、补肾3组,补肾组灌服左归丸浓缩剂。取各组新生仔鼠颅盖骨制取培养成骨细胞,观察细胞生长形态、ALP活性测定、RT-PCR检测第7、14、21天Cx43和BGPmRNA表达情况、免疫荧光染色检测第21天Cx43蛋白表达。结果:模型组仔鼠成骨细胞培养第5、7天ALP活性降低;第14、21天Cx43、BGPmRNA表达减少;第21天免疫荧光检测Cx43蛋白表达减少。上述指标与另两组比较差异显著(P<0.05)。结论:先天肾虚导致仔鼠颅骨成骨细胞分化成熟过程中Cx43、BGP表达减少,这是其影响骨早期发育的重要机制之一。左归丸补肾填精治疗可提高Cx43、BGP表达,保证成骨细胞正常功能,促进骨发育。

Objective ： Start with the effect of the congenital deficiency of kidney on the expression of Cx43 and BGP in the filial rats calvarial osteoblasts, and reserch the mechanism of its influence on the early bone development and demonstrate the function of tonifying kidney Tianjing therapy. Methods ： The model of congenital deficiency of kidney was induced by simulated earthquake vibration and electric stimulation combined with light which can scare pregnant rats. The pregnant rats were divided into normal group, model group and tonifying kidney group. The tonifying kidney group were intragastrically administered with Zuogui Pill concentrated mixture. We took each newborn rat＇s calvaria and prepared the cultured osteoblasts. Cell morphology was observed and then we detected the activity of ALP. RT - PCR method was used to detect the expressions of Cx43 and BGP mRNA on the 7th,14 th and 21st days. Cx43 protein expression was detected by immunofluorescence staining at day 21. Results ： In model group rats, the activity of ALP was decreased at day 5 and 7. The expressions of Cx43 and ALP mRNA decreased at day 14 and 21. The expression of Cx43 protein was decreased at day 21. Compared with the other two groups, the above indices of model group were significantly different （ P 〈 0. 05）. Conclusion ： The Cx43 and BGP expressions are decreased in the course of the differentiation and maturation of calvaria osteoblasts of newborn rats induced by congenital deficiency of kidney, which is one of the important mechanisms that affects the early development of bone. Tonifying kidney treatment by Zuogui Pill can increase the expressions of Cx43 and BGP, ensure the normal function of osteoblast, and promote the development of bone.