摘要
目的:探讨补中益气汤通过脂联素受体1(Adipor1)/磷酸腺苷活化蛋白激酶(AMPK)/过氧化物酶体增生物激活受体γ共激活因子-1α(PGC-1α)对运动性疲劳(EIF)小鼠骨骼肌中脂代谢的影响。方法:将C57BL/6J小鼠随机分为空白组、模型组、补中益气汤低、中、高剂量组、维生素C组。空白组不施加任何干预,其余组通过力竭游泳建立EIF模型。力竭游泳前1 h,空白组、模型组灌服0.2 mL蒸馏水,补中益气汤低、中、高剂量组分别灌胃4.1、8.2、16.4 g·kg^(-1)的补中益气汤,维生素C组灌胃0.04 g·kg^(-1)剂量维生素C,持续6周。实验6周后,计算小鼠体质量增长率、脏器指数、力竭游泳时间;酶比色法检测小鼠血清中尿素氮(BUN)、肌酸激酶(CK)、乳酸脱氢酶(LDH)、乳酸(LD)的水平;苏木素-伊红(HE)染色观察骨骼肌组织病理变化;透射电镜(TEM)观察骨骼肌组织超微结构;微板法检测血清中游离脂肪酸(NEFA)、甘油三酯(TG)的含量;蛋白免疫印迹法(Western blot)测定骨骼肌中Adipor1、磷酸化(p)-AMPK/AMPK、PGC-1α和己糖激酶2(HK2)蛋白的表达。结果:与空白组比较,模型组小鼠的体质量增长率、胸腺指数降低,血清中BUN、CK、LD、LDH水平升高(P<0.01),NEFA、TG含量下降(P<0.01),骨骼肌组织Adipor1、p-AMPK/AMPK、PGC-1α、HK2蛋白表达下降(P<0.05,P<0.01)。与模型组比较,补中益气汤高剂量组体质量增长率、胸腺指数、力竭游泳时间显著升高(P<0.01),BUN、CK、LD、LDH显著下降(P<0.01),NEFA、TG显著升高(P<0.01),骨骼肌组织Adipor1、p-AMPK/AMPK、PGC-1α、HK2蛋白表达明显升高(P<0.05,P<0.01);维生素C组胸腺指数、力竭游泳时间明显提高,BUN、CK、LD明显下降(P<0.05,P<0.01),NEFA、TG显著升高(P<0.01),骨骼肌组织Adipor1蛋白表达显著升高(P<0.01)。结论:补中益气汤能够延缓EIF的发生,其机制可能与调节Adipor1/AMPK/PGC-1α信号通路,提高骨骼肌对脂肪的利用率有关。
Objective:To discuss the impact of Buzhong Yiqitang on lipid metabolism in skeletal muscle of exercise-induced fatigue(EIF)mice through adiponectin receptor 1(Adipor1)/adenosine 5'-monophosphate(AMP)-activated protein kinase(AMPK)/peroxisome proliferator-activated receptor gamma coactivator-1α(PGC-1α).M ethod::C57BL6J mice were randomly divided into the control group,model group,low,middle,and high dose groups of Buzhong Yiqitang,and vitamin C group.No intervention was given to the control group,while the other groups were subjected to exhaustive swimming training to establish the EIF model.One hour before exhaustion,0.2 mL distilled water was given to the control group and the model group,while the mice in the low,middle,and high dose groups of Buzhong Yiqitang were given intragastrically Buzhong Yiqitang of 4.1,8.2,and 16.4 g·kg^(-1),respectively,and the vitamin C group was given vitamin C of 0.04 g·kg^(-1) via gavage for a duration of six weeks.After six weeks of the experiment,the growth rate of body weight,organ index,and exhaustive swimming time were calculated.Enzyme colorimetry was utilized to detect the levels of blood urea nitrogen(BUN),creatine kinase acid(CK),lactate dehydrogenase acid(LDH),and lactic acid(LD).The pathological changes of skeletal muscle were observed using hematoxylin-eosin(HE)staining,while the ultrastructure of skeletal muscle was observed with transmission electron microscope(TEM).The contents of free fatty acids(NEFA)and triglyceride acid(TG)in serum were also examined by microplate method.The protein expressions of Adipor1,p-AMPK/AMPK,PGC-1α,and HK2 in the skeletal muscle were measured by Western blot.Result::Compared with those of the control group,the growth rate of body weight and thymus index of the model group were decreased,and the serum levels of BUN,CK,LD,and LDH were increased(P<0.01).The contents of NEFA and TG were decreased(P<0.01),and the protein expression of Adipor1,p-AMPK/AMPK,PGC-1α,and HK2 in the skeletal muscle decreased(P<0.05,P<0.01).Compared with those in the model group,the growth rate of body weight,thymus index,and exhaustive swimming time were significantly increased(P<0.01),and the levels of BUN,CK,LD,and LDH dropped in the high dose group of Buzhong Yiqitang(P<0.01).The levels of NEFA and TG were greatly improved(P<0.01).The protein expressions of Adipor1,p-AMPK/AMPK,PGC-1α,and HK2 in the skeletal muscle were significantly increased(P<0.05,P<0.01).Compared with those in the model group,the thymus index and exhaustive swimming time were significantly increased in the vitamin C group,and the levels of BUN,CK,and LD dropped(P<0.05,P<0.01).The levels of NEFA and TG were improved significantly(P<0.01),and the protein expression of Adipor1 in skeletal muscle was increased greatly(P<0.01).Conclusion::Buzhong Yiqitang can delay the development of EIF,which may be connected with the regulation of the Adipor1/AMPK/PGC-1αsignaling pathway and the improvement of the utilization rate of skeletal muscle to fat.
作者
彭娟
韦华妮
姜婷
刘伟
潘华山
陈道睿
PENG Juan;W EI Huani;JIANG Ting;LIU Wei;PAN Huashan;CHEN Daorui(School of Physical Education and Health,Guangzhou University of Chinese Medicine,Guangzhou 510006,China;Lingnan Medical Research Center,Guangzhou University of Chinese Medicine,Guangzhou 510405,China;Guangdong Chaozhou Health Vocational College,Chaozhou 521000,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2024年第12期24-30,共7页
Chinese Journal of Experimental Traditional Medical Formulae
基金
广东省自然科学基金面上项目(2024A1515012151)
广东省方证研究重点实验室(2022B1212010012)
广东省普通高校特色创新类项目(2023KTSCX391)
广东省教育厅普通高校认定类科研项目(2021KTSCX237)。