Ginsenoside Rg3 Relieve Cancer-related Fatigue By Activating AMPK

Background and Objectives Cancer-related fatigue is a common complication of cancer patients.It is mainly manifested as fatigue,lack of concentration,etc.These symptoms are not related to the amount of exercise,but seriously affects the life quality of patients.Skeletal muscle fatigue is one of the key features of cancer-related fatigue.Inflammatory effects can damage mitochondrial function in some cancers and cancer treatments.Fatigue is related to energy metabolism and decreased mitochondrial efficiency in skeletal muscle.AMP-dependent protein kinase is the key molecule in the biological energy metabolism.It restores energy balance during metabolic stress in cellular and physiological.AMPK activation can increase PGC-1? gene expression or directly phosphorylate PGC-1? in skeletal muscle,thereby increasing mitochondrial biogenesis and mitochondrial metabolism.Anti-depressant drugs are mainly used in the clinical treatment of cancer-related fatigue.Such drugs have problems such as not obvious anti-fatigue effect and significant side effects.Ginseng is a traditional and valuable medicinal material in China.In recent years,research on ginsenoside-Rg3 has been gradually deepened.In addition to its anti-tumor properties,toxicity-reducing of chemotherapy drugs,ginsenoside-Rg3 also protects the central nervous system and cardiovascular system,resists fatigue,lowers blood sugar,and repairs wounds Extensive pharmacological effects.This project is to study the ginsenoside Rg3 effect of anti-cancer-related fatigue and its possible molecular mechanism.Method 1.A fatigue model was established in vitro,using lipopolysaccharide and preconditioning mouse myotube C2C12 cells and mouse primary myotube to mimic the cancer-induced fatigue microenvironment.2.Western blot,RT-q PCR,and immunofluorescence were used to detect the levels of AMPK? phosphorylation,mitochondrial production,and mitochondrial biomarker expression in mouse C2C12 myotube and mouse primary myotube in each group.3.Mito-green fluorescent was used to detect the number and activity of mitochondria in the muscle cells.4.Cancer-related fatigue mouse model was established,we randomly divided the mice into four groups: a control healthy group,a tumor-bearing group,a tumorbearing intraperitoneal injection of Rg3 group,and a tumor-bearing Rg3 intragastric group,8 mice in each group.The mice were weighed every two day,and tumor volume was measured every three day after tumor formation(v = a × b2/2).The fatigue of mice in each group was measured using the platform treadmill test.After 7 days,two drug treatment groups were administered once a day at a dose of 5mg/kg for 14 consecutive days.5.Western blot,RT-q PCR,and immunofluorescence were used to detect the AMPK? phosphorylation level,mitochondrial production and mitochondrial biomarker expression levels in the thigh muscles of mice in each group.Result1.Rg3 treatment of mouse C2C12 myotube and primary myotube increased the level of AMPK? phosphorylation in medicine concentration and time-dependent manner.2.Treating with 2ng/ml LPS in mouse myotube formed cancer-induced fatigue microenvironment.Rg3 increased phosphorylation of intracellular AMPK?.3.In vitro cancer-related fatigue microenvironment,Rg3 increased mitochondrial biosynthesis.The expression of mitochondrial biosynthesis proteins increased under Rg3 treatment.4.In vitro cancer-related fatigue microenvironment,Rg3 increased mitochondrial metabolic activity by activating AMPK?.After treatment with Rg3 under fatigue environment,mitochondrial activity was enhanced,and the expression of mitochondrial metabolism-related proteins Cytochrome C and COX IV was increased.5.In the cancer-related fatigue mouse model,the mice in the tumor-bearing group had shorter running distances and slower running speeds on the treadmill test.In the treadmill test,the mice in the tumor-bearing group had longer running distance and faster speed.6.Western blot,RT-q PCR,tissue immunofluorescence,and immunohistochemical results showed that the AMPK? phosphorylation level in the skeletal muscle tissue of the Rg3 group(either the intraperitoneal injection group or the intragastric administration group)was higher than the tumor-bearing group,while the mitochondrial biosynthesis and mitochondrial metabolism-related markers PGC-1?,NRF-1,TFAM,Cytochrome C,COX IV levels was increased.Conclusion 1.Rg3 increased the level of AMPK? phosphorylation in primary myotube and mouse C2C12 myotube.2.Rg3 increased mitochondrial biosynthesis by activating mouse C2C12 myotube and primary myotube AMPK?.3.Rg3 enhanced mitochondrial metabolism by activating mouse C2C12 myotube and primary myotube AMPK?.4.Rg3 relieved fatigue-like behavior in mouse models of cancer-related fatigue.5.Rg3 released the fatigue-liked symptoms of cancer-related fatigue by increasing the level of AMPK? phosphorylation,increasing the mitochondrial biosynthesis capacity and mitochondrial metabolic activity in cancer-related fatigue mice.