Extraction Of Cordyceps Flower Polysaccharides And The Protective Effect On CCl₄-induced Mice Liver Fibrosis

Liver fibrosis is caused by excessive accumulation of extracellular matrix?ECM? during chronic liver diseases such as viral hepatitis,cholestasis and non-alcoholic steatohepatitis.Advanced liver fibrosis can lead to severe liver diseases,including cirrhosis,liver failure and portal hypertension.The pathological mechanism of liver fibrosis is complex,and there are no drugs with definite efficacy for clinical application at present.Natural products can treat liver fibrosis through multiple targets,which has become a hotspot in the treatment of liver fibrosis in recent years.Cordyceps militaris?Lex.Fr?Link cultivated solely on artificial medium was called cordyceps flower.Studies have shown that Cordyceps militaris polysaccharides have biological activities such as antioxidant and hypoglycemic.Cordyceps flower has a lower price and higher polysaccharide content compared with Cordyceps militaris,indicating a higher application value.In order to explore the optimal extraction process of cordyceps flower polysaccharides?CFP?,we optimized polysaccharide extraction process by response surface analysis,and removed protein by Sevag method to obtain CFP at a purity of 95.61%.In order to explore the role of CFP on liver fibrosis and the involved mechanisms,we used CCl₄-induced liver fibrosis mice model in our study.At the end of experiment,we collected mice serum and liver tissues for the investigation.By completing our experiments,we can draw the following conclusions:?1?Response surface analysis based on the single factor experiment was designed to optimize the extraction process of CFP.The best process parameters are:extraction temperature at 87?,the water to raw material ratio was 41:1?m L/g?,and the extraction time was 170 min.Under this condition,the yield of CFP was?6.21±0.22?%.?2?In vitro,we found that CFP have no significant cytotoxicity.?3?Bodyweight is an important health indicator.We found that CCl₄ administration caused bodyweight loss,while feeding with CFP significantly increased the lost bodyweight,indicating that CFP can improve the toxicity caused by CCl₄.The measurement of serum AST and ALT showed that feeding of CFP significantly reduced CCl₄-upregulated AST and ALT levels.H&E staining of liver tissue sections indicated that CCl₄ administration resulted in the necrosis of hepatocytes and the denaturation of hepatic lobules while CFP treatment alleviated these injuries.These results collectively indicated that CFP can alleviate CCl₄-induced liver injury.?4?We used Sirius red staining to detect liver fibrosis,and the results showed that CFP significantly reduced collagen content caused by CCl₄.In addition,compared with model mice,the expression of fibrosis-related proteins type I collagen?collagen-1?,transforming growth factor-?1?TGF-?1?,and?-smooth muscle actin??-SMA?was inhibited in GFP treated mice.These results showed that CFP alleviates liver fibrosis in mice.?5?Inflammation plays an important role in liver fibrosis.We found that nuclear factor?B?NF-?B?and tumor necrosis factor??TNF-??levels were inhibited by CFP,which were induced by CCl₄.In addition,we also found CFP increased CCl₄ inhibited peroxisome proliferators-activated receptor??PPAR??expression,another important nuclear transcription factor for inflammation regulation.These results indicate that CFP alleviate liver fibrosis by inhibiting inflammation.?6?Furthermore,our results showed that CCl₄ reduced B-cell lymphoma-2?BCL-2? and increased BCL2-associated X protein?BAX?expression in mice liver.However,CFP significantly enhanced BCL2 and inhibited BAX levels.These results indicate that CFP improves liver fibrosis by inhibiting CCl₄-induced hepatocytes apoptosis.In summary,we demonstrate that CFP can alleviate liver fibrosis in mice.In mechanisms,we showed that CFP can protect liver by inhibiting inflammation and apoptosis.Our study provides a new therapy for liver fibrosis.