世界中医药

作者	单位
王康1，孟凤仙1，钮妍2，薛恬珏3，国生1，沈潜1，魏培栋1，付国兵1	1 北京中医药大学东方医院，北京，100078； 2 北京中医药大学第三附属医院，北京，100029； 3 北京中医药大学，北京，100029

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中文摘要:

目的：观察推拿对2型糖尿病（Type 2 Diabetes Mellitus，T2DM）大鼠腹直肌组织腺苷酸活化蛋白激酶α2(AMPKα2）、核因子-κB（Nuclear Factor-κB，NF-κB）及白细胞介素-6(Interleukin-6，IL-6）基因转录及蛋白表达的影响，探讨其改善T2DM大鼠代谢性炎性损伤的分子机制。方法：选取40只Wistar大鼠，随机取出6只为空白组，其余高脂高糖饮食联合低剂量（35 mg/kg）链脲佐菌素腹腔注射建立T2DM模型，成模大鼠32只随机分入模型组、西药组、推拿组及西药推拿组。空白组及模型组每日灌胃蒸馏水，西药组每天以250 mg/kg标准进行二甲双胍灌胃1次，推拿组每日给予推拿手法治疗1次，西药推拿组在相同给药标准基础上每日加用推拿治疗1次。共干预8周，分别记录第2、4、8周的空腹血糖（Fast Blood Glucose，FBG）值，并运用realtime-PCR测定腹直肌AMPKα2、NF-κB及IL-6基因转录，Western-blot测定腹直肌AMPKα2及NF-κB蛋白含量，ELISA测定IL-6蛋白含量。结果：与模型组比较，西药组及西药推拿组FBG在第2、4、8周时显著下降，差异有统计学意义（P<0.05，P<0.01），推拿组FBG在第4、8周时显著下降，差异有统计学意义（P<0.05）；与西药组比较，西药推拿组在第4周降糖效果更为显著，差异有统计学意义（P<0.05）；与推拿组比较，西药推拿组在第4、8周时降糖效果更为显著，差异有统计学意义（P<0.05）；与0周时比较，各观察组FBG在第2、4、8周时，均显著降低，差异有统计学意义（P<0.05，P<0.01）。与空白组比较，模型组AMPKα2mRNA转录及蛋白表达均下降，差异有统计学意义（P<0.05）；与模型组比较，西药组、推拿组、西药推拿组AMPKα2mRNA转录及蛋白表达均升高，差异有统计学意义（P<0.05）。与空白组比较，模型组NF-κBmRNA转录显著升高，差异有统计学意义（P<0.01），蛋白表达升高，差异有统计学意义（P<0.05）；与模型组比较，西药组NF-κBmRNA转录下降，差异有统计学意义（P<0.05）；与模型组比较，推拿组NF-κBmRNA转录及蛋白表达均下降，差异有统计学意义（P<0.05）；与模型组比较，西药推拿组NF-κBmRNA转录和蛋白表达显著下降，差异有统计学意义（P<0.01，P<0.05）；与空白组比较，模型组IL-6 mRNA转录升高，差异有统计学意义（P<0.05），蛋白表达显著升高，差异有统计学意义（P<0.01）；与模型组比较，西药组、推拿组及西药推拿组IL-6基因转录均显著下降，差异有统计学意义（P<0.05，P<0.01），蛋白表达均显著下降，差异有统计学意义（P<0.01）。结论：推拿可改善T2DM大鼠的空腹血糖，并通过上调骨骼肌AMPKα2基因转录与蛋白表达，下调NF-κB、IL-6基因转录与蛋白表达，发挥其改善糖脂代谢性炎性损伤的手法效应。

English Summary:

To observe the effects of Tuina on the gene transcription and protein expression of 5'AMP-activated protein kinaseα2 (AMPKα2)，nuclear factor-κB (NF-κB) and interleukin(IL-6) in rectus abdominis muscle of type 2 Diabetes Mellitus (T2DM) rats.To explore the molecular mechanism of Tuina on improving the metabolic inflammatory injury in T2DM.Methods:A total of 6 Wistar rats out of 40 were randomly selected as the blank group (CON).Others were feed with high-fat and high-sugar diets combined with low-dose (35 mg/kg) streptozotocin to establish a T2DM model.A total of 32 successfully modeled rats were randomly divided into the model group (MOD)，western medicine group (MET)，and Tuina group (TNA) and western medicine combined Tuina group (MIX).The CON and the MOD were perfused with distilled water daily.The MET was daily given a dose of 250 mg/kg for metformin，and the TNA was given daily for manual treatment，and the MIX was given both intervention from MET and TNA.After 8 weeks of intervention，the values of fast blood glucose (FBG) at 2nd，4th，and 8th week were recorded，and real-time PCR was used to measure the gene transcription of AMPKα2，NF-κB，and IL-6 in the rectus abdominis.Western-blot was used to measure the protein expression of AMPKα2 and NF-κB in the rectus abdominis muscle，and the protein expression of IL-6 was determined by ELISA.Results:Compared with the MOD，the FBG of the MET and the MIX decreased significantly at the 2nd，4th，and 8th week (P<0.05，P<0.01)，and the FBG of the TNA decreased significantly at the 4th and 8th week (P<0.05).Compared with the MET，the MIX had a more significant hypoglycemic effect at the 4th week (P<0.05)，and compared with the TNA，the MIX had a more significant hypoglycemic effect at the 4th and 8th week (P<0.05).Compared with 0 weeks，FBG of each treatment group was significantly decreased at the 2nd，4th，and 8th week (P<0.05，P<0.01).Compared with the CON，the mRNA and protein expression of AMPKα2 in the MOD was decreased (P<0.05).Compared with the MOD，the mRNA and protein expression of AMPKα2 in the MET，the TNA and the MIX were increased (P<0.05).Compared with the CON，the transcription of NF-κB mRNA in the MOD was significantly increased (P<0.01)，and the protein expression was increased (P<0.05).Compared with the MOD，the transcription of NF-κB mRNA in the MET was decreased (P<0.05).Compared with the MOD，the transcription of NF-κB mRNA in the MET was significantly decreased (P<0.01)，and the protein expression was decreased (P<0.05).Compared with the CON，the IL-6 mRNA expression in the MOD was increased (P<0.05)，and the protein expression was significantly increased (P<0.01).Compared with the MOD，the IL-6 gene transcription in the MET，the TNA and the MIX was all significantly decreased (P<0.05，P<0.01)，and protein expression was also decreased significantly (P<0.01).Conclusion:Tuina can improve the FBG of T2DM rats，and up-regulate the gene transcription and protein expression of AMPKα2 in skeletal muscle，down-regulate the gene transcription and protein expression of NF-κB and IL-6，in order to improve the metabolic damage caused by glycolipid toxicity.

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