“脾肾相赞”理论探讨不同治法对大鼠骨髓间充质干细胞成肌分化中TNF-α、JNK含量影响  

Based on the theory of"praise of spleen and kidney",the effects of different treatment methods on TNF-αand JNK content in myogenic differentiation of BMSCs

作　　者：刘彤[1] 李俊儒 付夜平 王致远 林庶茹[1] 杨芳[1] LIU Tong;LI Junru;FU Yeping;WANG Zhiyuan;LIN Shuru;YANG Fang(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China)

机构地区：[1]辽宁中医药大学,辽宁沈阳110847

出　　处：《中国骨质疏松杂志》2024年第4期469-473,共5页Chinese Journal of Osteoporosis

基　　金：国家重点研发计划项目(2018YFC1704301);国家自然科学基金面上项目(81973719);辽宁“百千万人才工程”培养经费资助(2020921100)。

摘　　要：目的在“脾肾相赞”理论指导下,比较不同中医治法对大鼠BMSCs成肌分化及过程中TNF-α、JNK蛋白表达的影响,分析中医治疗骨质疏松症的机制。方法将大鼠分为正常组、诱导组、补肾组、健脾组、补肾健脾组(随机数字表法),制备含药血清,选取第4代BMSCs进行实验,运用免疫荧光法分析各组成肌分化情况,酶联免疫吸附法检测TNF-α、JNK蛋白表达量。结果①BMSCs经诱导15d后,通过免疫荧光化学染色法检测MyoG荧光平均强度,结果显示:各组均显著高于正常组(P<0.01),补肾健脾组最高。②TNF-α蛋白检测:补肾组、补肾健脾组显著低于正常组(P<0.01)。③JNK蛋白检测:各组显著低于正常组(P<0.01)。结论①各治法中,补肾健脾法对大鼠BMSCs成肌分化的促进效果最佳,为通过脾肾、肌骨双方向治疗OP提供可能。②各中医治法均对大鼠BMSCs的成肌分化存在促进作用,这可能与降低TNF-α、JNK水平有关。Objective Under the guidance of the theory of"praise of spleen and kidney",the effects of different TCM treatments on the myogenic differentiation of rat BMSCs and the presence of TNF-αand JNK protein were compared,and the mechanism of TCM in treating osteoporosis was analyzed.Methods Randomly dividing rats into the normal group,the induction group,the kidney-tonifying group,the spleen-invigorating group and the kidney-tonifying and spleen-invigorating group.The fourth generation BMSCs was selected for experiment.The immunofluorescence technique was used to analyze the muscle differentiation in each group and the enzyme-linked immunosorbent assay(Elisa)was employed to detect the presence of TNF-αprotein and JNKprotein.Results①The results showed that,following a 15-day induction with BMSCs,the immunofluorescence chemical stainingrevealed an average fluorescence intensity of MyoG.In comparison to the normal group,the average the average fluorescence intensity of MyoG in other groups was significantly higher than that in the normal group,particularly in the kidney-tonifying and spleen-invigorating group.②In comparison to the normal group,the presence of TNF-αproteinin the kidney-tonifying group and the kidney-tonifying and spleen-invigorating group was significantly lower(P<0.01).③In comparison to the normal group,the presence of JNK protein in other groups was significantly lower(P<0.01).Conclusion①Among the treatment methods of TCM,the method of tonifying the kidney and invigorating the spleen is the best in promoting the myogenic differentiation of rat BMSCs,which provides the possibility for the treatment of OP through both spleen and kidney,muscle and bone.②All TCM treatments can promote the myoblast differentiation of BMSCs in rats,which may be related to the decrease of the levels of TNF-αand JNK.

关 键 词：脾肾相赞 骨髓间充质干细胞 成肌分化 TNF-Α JNK 

分 类 号：R681[医药卫生—骨科学]