独活寄生汤对大鼠腰椎间盘细胞活性及CaM-CaMK-CREB信号通路的影响
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1. 福建省立医院,福建 福州 350003;2. 福建中医药大学附属第二人民医院,福建 福州 3500033. 福建生物工程职业技术学院,福建 福州 350001

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R285.5

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基金项目: 福建省教育厅课题(JB11084);福建省中医处中医药科研课题(wzkf201304/wzkf201302) 收稿日期: 2015 - 02 - 23 作者简介: 钟灼琴(1985-)女,福建罗源人,住院医师,在读硕士研究生,主要研究方向:脊椎病康复。△通信作者:张坤木,E-mail:277873495@qq.com


Effect of Duhuo Jisheng Decoction on the Cells Activity and Signal Transductions of CaM-CaMK-CREB Occurring in Lumbar Disc Cells of Adult Rats
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1. Fujian Provincial Hospital, Fuzhou 350003, China;2. Second People’s Hospital, Fujian University of Traditional Chinese Medicine, Fuzhou 350003, China;3. Fujian Vocational College of Bioengineering, Fuzhou 350001, China

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    摘要:

    目的研究独活寄生汤治疗腰椎间盘突出症与CaM-CaMK-CREB信号通路的内在关联。方法用独活寄生汤中药血清、空白血清及胎牛血清对大鼠腰椎间盘纤维环细胞进行分组干预。采用MTT比色法检测独活寄生汤中药血清作用后的纤维环细胞生长曲线;应用western blotting技术检测独活寄生汤中药血清对CaM、CaMK、CREB表达及定量的影响,并对检测结果进行数据处理及统计学分析。结果中药血清干预后的第1天,5组之间两两比较无明显差异(P>0.05);中药血清干预后的第2天至第7天各组与空白血清组比较均有明显差异(P<0.05);与空白血清组比较,各剂量独活寄生汤中药血清均能可显著提高大鼠腰椎间盘纤维环细胞内的CaM、CaMKII、CaMKIV和CREB等蛋白的表达水平(P<0.01),且随着中药剂量的增加CaM、CaMKII、CaMKIV和CREB等蛋白的表达水平提升能力也在增加。结论独活寄生汤中药血清可明显促进大鼠椎腰间盘纤维环细胞生长,随着中药剂量的增加中药血清促进细胞生长能力增强;临床上独活寄生汤治疗腰椎间盘突出症效应过程可能存在一条重要的信号通路“CaM-CaMK-CREB”。

    Abstract:

    Objective The relationship between the CaM-CaMK-CREB signal pathway and the treatment for lumbar disc disease by Duhuo Jisheng decoction was studied. Methods Group intervention on fibrous ring cells of lumbar intervertebral disc of adult rats by medicine serum mixed with Du huo ji sheng decoction, blank serum and fetal calf serum respectively was made. Detect by MTT colorimetric assay the growth curve of fibrous ring cell under the adoption of Du huo ji sheng decoction serum;Conduct western blotting to detect the impact of herbal medicine serum on expression and quantification of CaM, CaMK and CREB;Perform data processing and statistical analysis on the results. Results The five groups showed no significant difference between each other during the first day after the intervention of Chinese medicine serum(P>0.05); from the second to seventh day after the intervention, there were significant difference between the blank serum group and other groups(P<0.05). Compared with the blank serum group, Du huo ji sheng Decoction medicine serum at all doses could substantially improve the expression of CaM, CaMKII, CaMKIV and CREB in fibrous ring cells of lumbar intervertebral disc of adult rats(P<0.01), and with the increased doses of Chinese medicine, the capacity of improving the expression level was also increasing. Conclusion Du huo ji sheng decoction medicine serum could highly stimulate the development of fibrous ring cells of lumbar intervertebral disc of adult rats. Clinically when it comes to the treatment by Du huo ji sheng decoction for lumbar disc disease, there is likely to be an important signal pathway called “CaM-CaMK-CREB”.

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