中医不同治法调控ERK2、JNK促进肌源性干细胞向成骨分化的研究

目的 通过观察中医不同治法促进大鼠肌源性干细胞(muscle-derived stem cells, MDSCs)成骨分化及细胞外调节蛋白激酶2(extracellular regulated protein kinase 2,ERK2)、c-Jun氨基末端激酶(c-Jun N-terminal kinase, JNK)蛋白表达情况,探究基于“脾肾相关”理论下,中医防治骨质疏松症(osteoporosis, OP)的作用机制。方法 在随机数字表法下,将大鼠分为正常组、诱导组、补肾组、健脾组及补肾健脾组,制备含药血清,选取第4代大鼠MDSCs进行实验,CCK8法检测细胞增殖,p-NPP法检测细胞碱性磷酸酶(alkaline phosphatase, ALP)活性,ELISA法检测细胞ERK2、JNK蛋白表达。结果 (1)各组均可促进MDSCs增殖。(2)与正常组相比,各组ALP活性显著升高(P<0.01),其中补肾健脾组最高。(3)诱导组、补肾组、补肾健脾组ERK2蛋白表达显著高于正常组(P<0.01),健脾组高于正常组,差异无统计学意义;各组JNK蛋白表达显著低于正常组(P...

Effects of different treatments of TCM on osteogenic differentiation by muscle-derived stem cells through regulation of ERK2 and JNK

Objective By observing the osteogenic differentiation of rat muscle-derived stem cells (MDSCs) induced by different treatments of traditional Chinese medicine and the expression of extracellular regulated protein kinase 2 (ERK2) and c-Jun N-terminal kinase (JNK), to discuss the mechanism of traditional Chinese medicine in the prevention and treatment of osteoporosis (OP) under the correlation between the spleen and kidney theory. Methods Using the random number table method, rats were divided into the normal group, the induction group, the kidney-tonifying group, the spleen-invigorating group, the kidney-tonifying, and the spleen-invigorating group. Drug-containing serum was prepared and the fourth generation of rat MDSCs were selected for the experiment. The proliferation of MDSCs was measured using CCK8 method. The activity of alkaline phosphatase (ALP) was detected using p-NPP method. The protein expressions of ERK2 and JNK were measured using ELISA. Results (1) The proliferation of rat MDSCs was promoted in each group. (2) Compared to that in the normal group, the activity of ALP in other groups increased significantly (P<0.01), and it was the highest in the kidney-tonifying and the spleen-invigorating group. (3) The protein expression of ERK2 in the induction, the kidney-tonifying, the kidney-tonifying, and the spleen-invigorating group was much higher than that in the normal group (P<0.01), but the increase in the spleen-invigorating group was not obvious. The protein expression of JNK in other groups was much lower than that that in the normal group (P<0.01). Conclusion (1) The osteogenic differentiation by rat MDSCs may promot by tonifying the kidney and invigorating the spleen, indicating that under the guidance of spleen and kidney correlation theory, strengthening the role between muscle and bone may prevent and treat osteoporosis. (2) The change of protein expression of ERK2 and JNK suggests that the mechanism of traditional Chinese medicine in the treatment of osteoporosis may be related to the regulation of the expression of ERK2 and JNK protein, which provides a new way for traditional Chinese medicine in the clinical diagnosis and treatment of osteoporosis.