补肾健脾活血方调节尾部悬吊大鼠肌蛋白合成与分解的机制研究  

Study on the Mechanism of Bushen Jianpi Huoxue Decoction(补肾健脾活血方)in Regulating the Synthesis and Decomposition of Muscle Protein in Tail-suspended Rats

作　　者：王萌[1] 李盼盼[1] 韩行 张林[1] 谢鑫[1] 张红梅[1] 卢健[1] WANG Meng;LI Panpan;HAN Hang;ZHANG Lin;XIE Xin;ZHANG Hongmei;LU Jian(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China)

机构地区：[1]辽宁中医药大学,辽宁沈阳110847

出　　处：《辽宁中医药大学学报》2023年第5期38-42,共5页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81774246);辽宁省自然科学基金指导计划项目(LJKQZ2021065)。

摘　　要：目的探究补肾健脾活血方对废用性肌萎缩的影响及过程中肌蛋白合成与分解机制。方法选取40只2月龄雌性大鼠,分为5组:空白组(A)、造模组(B)、低药量组(C)、中药量组(D)、高药量组(E),每组8只。除A组自由活动外,其余4组按照尾部悬吊法进行造模。C、D、E组给予不同剂量补肾健脾活血汤干预,A、B组给予等量蒸馏水。给药28 d后处死大鼠,称重比目鱼肌,测量湿重/体质量比。采用HE染色法对比目鱼肌肌肉组织形态与细胞核聚集状态进行病理形态学观察。ELISA检测补肾健脾活血方对血清中肌肉生长抑制素(myostatin,MSTN)的干预情况。Western blot检测补肾健脾活血方对比目鱼肌肌肉组织中丝氨酸/苏氨酸蛋白激(AKT)、SMAD蛋白3(Smad3)、叉头蛋白转录因子(FOXO1)、肌萎缩F-box蛋白(MAFbx)、肌环指蛋白-1(MuRF1)蛋白表达的干预情况。结果形态学观察显示,同A组相比,B组大鼠比目鱼肌湿重/体质量比值显著降低(P<0.01),肌纤维横截面明显减少,肌纤维密度明显增加,肌纤维间筋膜横截面明显增加,肌束间筋膜横截面明显减少,细胞核高度聚集;同B组比较,C、D、E组肌肉湿重/体质量比值有所上升(P<0.05),肌纤维横截面明显增加,肌纤维密度明显降低,肌束间筋膜横截面显著增加,核聚集程度降低;同D组比较,C组和E组肌肉湿重/体质量比值无明显差异(P>0.05),但伴随药物浓度的增加,C、D、E组肌肉湿重/体质量比值呈现逐渐增加趋势,且肌纤维横截面逐渐增大,肌纤维密度逐渐减小,肌束间筋膜横截面增加逐渐显著,核聚集减少逐渐显著。ELISA检测发现,同A组相比,B组大鼠血清中的MSTN水平显著升高(P<0.01);同B组相比,C组MSTN水平下降(P<0.05),D、E组MSTN水平明显下降(P<0.01);同D组比较,C、E组MSTN水平没有显著性差异(P>0.05)。Western blot检测结果显示,同A组相比,B组大鼠体内Smad3、Akt蛋白表达量减少(P<0.05),FOXO1、MuRF1、MAFObjective To investigate the effect of Bushen Jianpi Huoxue Decoction(补肾健脾活血方)on disuse muscular atrophy and the mechanism of myoprotein synthesis and decomposition in the process.Methods Forty 2-month-old female rats were divided into five groups:blank group(A),model group(B),low dosage group(C),traditional Chinese medicine group(D),high dosage group(E),with 8 rats in each group.Excepting for group A,the other four groups were modeled according to the tail suspension method.C,D and E were intervened with different doses of Bushen Jianpi Huoxue Decoction,and groups A and B were intervened with the same amount of distilled water.After 28 days of administration,the rats were killed,the soleus muscle was weighed,and the wet weight/body mass ratio was measured.HE staining was used to compare the morphology of soleus muscle tissue and the state of nuclear aggregation for pathological observation.The intervention of Bushen Jianpi Huoxue Decoction on serum myostatin(MSTN)was detected by ELISA.Western blot was used to detect the intervention of Bushen Jianpi Huoxue Decoction on the expression of serine/threonine protein kinase(AKT),SMAD protein 3(Smad3),forkhead protein transcription factor(FOXO1),muscle atrophy F-box protein(MAFbx)and muscle ring finger protein-1(MuRF1)in soleus muscle tissue.Results Morphological observation showed that compared with group A,the ratio of muscle wet weight to body mass of soleus muscle in group B was significantly decreased(P<0.01),the cross section of muscle fiber was significantly decreased,the muscle fiber density was significantly increased,the cross section of intermuscle fiber fascia was significantly increased,the cross section of intermuscle fascicular fascia was significantly decreased,and the nuclei was highly aggregated.Compared with group B,the ratio of muscle wet weight to body mass in groups C,D and E were increased(P<0.05),the cross section of muscle fiber were significantly increased,the muscle fiber density were significantly decreased,the cross section of

关 键 词：废用性肌萎缩 补肾健脾活血方 肌蛋白分解 肌蛋白合成 

分 类 号：R285.5[医药卫生—中药学]