Total flavonoids of litchi seed inhibit HSC-T6 cells via MAPK signaling pathway
Objective It is to observe the effects of total flavonoids of litchi seed(TFLS)on the expression of MAPK sig-naling pathway related proteins in HSC-T6 cells,and to explore whether TFLS can reverse the process of liver fibrosis via reg-ulating MAPK signal pathway.Methods The TFLS groups of 20 μg/mL,40 μg/mL,80 μg/mL,160 μg/mL,320 μg/mL,cell control group and blank groups were set up,the absorbance of the cells in each group was detected by CCK-8 method to calculate cell proliferation inhibition rate,and the optimal concentration of TFLS was selected to carry out the subsequent experiments.Then TFL group,colchicine group,cell control group and blank group were set up and cultured in respective culture medium.After 24 h,48 h,72 h of culture,the absorbance of of cells in each group was detected by CCK-8 meth-od,and the cell proliferation inhibition rate was calculated;after 72 h of culture,the ultrastructure of cells was observed by transmission electron microscopy,the levels of matrix metalloproteinase-2(MMP-2),collagen typeⅠ(Col-Ⅰ),and col-lagen type Ⅲ(Col-Ⅲ)were detected by ELISA,the expressions of c-Jun N-terminal kinase(JNK),extracellular signal-regulated kinase(ERK),and p38 mRNA were detected by RT-PCR,and the expressions of JNK,ERK,p38 proteins and their corresponding phosphorylated kinase(p-JNK,p-ERK,and p-p38)in cells were detected by immunohistochemistry.Results The optimal concentration of TFLS was 160 μg/mL and the optimal intervention time was 72 h.Compared with the cell control group,the OD values of TFLS group and colchicine group after 24 h,48 h and 72 h of culture were significantly decreased(all P<0.05).After 72 h of culture in the TFLS group,the cell gap became larger,the cilia around the cell membrane dropped,the mitochondrial cristae in cytoplasm were broken or disappeared,some mitochondria were vacuo-lated,the flat pool of rough endoplasmic reticulum was dilated with attached ribosomes degranulated,the chromatin in nu-cleus was condensed with some margination and crumpled nuclear membrane,and formed apoptotic vesicles were found;the levels of MMP-2,Col-Ⅰ and Col-Ⅲ in the TFLS group and colchicine group were significantly lower than those in the cell control group(all P<0.05),and the level of MMP-2 in the colchicine group was significantly lower than that in the TFLS group(P<0.05);the relative expressions of JNK,ERK and P38 mRNA,and the relative expressions of p-JNK,p-ERK,p-P38 proteins in the TFLS group were significantly lower than those in the cell control group(all P<0.05),and none of the differences were statistically significant when compared with the colchicine group(all P>0.05).Conclusion TFLS has biological effects on liver fibrosis,and its mechanism may be related to regulating MAPK signaling pathway.
Total flavonoids of litchi seedliver fibrosisMAPK signaling pathwayhepatic stellate cells
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