Full
Outline
To study the molecular mechanism of Liujunzi Decoction（六君子汤）to improve C2C12 myoblast differentiation in tumor environment，providing experimental basis for the treatment of tumor cachexiamuscle atrophy with strengthening spleen and harmonizing stomachmethod.
It induced the differentiation of myoblasts C2C12 and observed the length of the differentiated myotube with an inverted microscope. Myoblasts C2C12 were co-cultured with lung cancer cell Lewis，and then were divided into blank group，model group，Fork head box protein O1（FoxO1）inhibitor group，Liujunzi Decoction（六君子汤）group and combination group. Western Blot detected the expression of myogenic differentiation antigen（MyoD）in C2C12 cells and the expressions of proteins associated with the peroxisome proliferator-activated receptor gamma coactivator 1-alpha（PGC-1α）/FoxO1 pathway. Electron microscopy observed the changes in the number of mitochondria and ultrastructure of C2C12 cells. Flow cytometry observed the changes inmitochondrial membrane potential of C2C12 cells. Quantitative real-time PCR detected mRNA in the PGC-1α/FoxO1 pathway in C2C12 cells.
Experiments showed that C2C12 cells had the longest transverse diameter in the muscle tube after 48 h of differentiation. Compared with those in the blank group，the MyoD protein expression and mitochondria number were significantly reduced in the model group（P<0.05）. The wrinkled boundary of themitochondrial bilayermembrane was not clear or even more C-shaped or stromal swelling，and the number of mitochondrial cristae was reduced. The arrangementwas disordered，fractured or dissolution disappeared into vacuoles. The mitochondrial membrane potential was decreased. The apoptotic ratewas significantly increased（P<0.05）. The mRNA expression ofmitochondrial transcription factor A（tFAM）was significantly reduced，and FoxO1 protein expression was significantly increased. The protein expressions of PGC-1α，FTO，mTOR and tFAM were significantly decreased（P<0.05）. Compared with themodel group，the expression of MyoD protein increased significantly in Liujunzi Decoction group（P<0.05）. In FoxO1 inhibitor group，Liujunzi Decoction group and the combination group，the number of mitochondria was significantly increased（P<0.05），the mitochondrial bilayer outermembrane structure was normal，the mitochondrial membrane potentialwas increased and the apoptosis rate was significantly reduced（P<0.05）. The mRNA expressions ofmTOR and tFAM in C2C12 cells weres significantly increased in the combination group（P<0.05），the expressions of FoxO1 protein in C2C12 cells in the FoxO1 inhibitor group，Liujunzi Decoction group and the combination group were significantly reduced，and the protein expressions of PGC-1α，FTO，mTOR and tFAM were significantly increased（P<0.05）. Compared with those in the Liujunzi Decoction group，the PGC-1αprotein expression in C2C12 cells in combination group significantly increased（P<0.05）.
Liujunzi Decoction can improve mitochondrial function and myoblast differentiation in tumor environment by interfering with PGC-1α/FoxO1 signal pathway.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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