Study On The Effect And Mechanism Of Wangshi Baochi Pills In Preventing Alcoholic Liver Disease

Background and PurposeAlcoholic liver disease(ALD)is one of the most common chronic liver diseases in the world,and there is no specific drug in clinic.It has been reported that ALD is closely related to acetaldehyde-induced toxicity,oxidative stress,hepatic steatosis,and intestinal flora dysregulation.WangshiBaochi(WSBC)pills are Chinese patent medicine which commonly used in clinical pediatrics.It is derived from a secret recipe created by Wang Luqing,a famous doctor of the Qing Dynasty.It is a state secret formula.WSBC pills can clear away heat,detoxify phlegm and disperse silt.WSBC pills are mainly used in the treatment of children’s anorexia,emesis,constipation,abdominal distension,chronic superficial gastritis and so on.There is no report of WSBC pills on the prevention and treatment of metabolic diseases.The purpose of this study is to explore the preventive and therapeutic effect of WSBC pills on ALD and its related mechanisms.Research ContentsThis study is mainly divided into two parts.In the first part,we create a model of acute alcoholic liver injury.We determine the activity and expression of Alcohol dehydrogenase(ADH)in mice with acute alcoholic liver injury and count the sober time of the drunk mice.The degree of liver injury was detected by pathological examination、serum aspartate transaminase(AST)and alanine aminotransferase(ALT).Through the detection of reactive oxygen species(ROS)and superoxide dismutase(SOD),through Western Blot analysis to detect the expression levels of SOD1,SOD2 and NADPH dehydrogenase quinone-1(NQO-1)to explore the mechanism of anti-oxidative stress of WSBC pills.In the second part,we establish a mouse model of chronic ALD induced by liquid alcohol and high fat diet.The activity and expression of ADH in ALD mice were determined.The pathological status of liver was investigated by pathological detection,serum AST and ALT detection.Through observing liver phenotype,oil red O staining and serum Triglyceride(TG)content detection to explore the degree of lipid accumulation in liver.The expressions of FASN,SREBP1,CD36,PPAR-α,CPT1α,IL-1α and TNF-α were detected by RT-PCR and Western Blot analysis.Through the establishment of chronic ALD model of antibiotic intervention,the role of intestinal flora in the prevention and treatment of ALD by WSBC pills was explored.Serum AST and ALT were detected to explore the pathological state of liver,liver lipid accumulation was detected by oil red O staining,and changes in the composition and quantity of intestinal flora were investigated by microbial diversity analysis experiment.ResultsIn the model of acute alcoholic liver injury,WSBC pills can increase the activity and expression of ADH in mice with acute alcoholic liver injury,and shorten the sober-up time of mice.WSBC pills can reduce liver injury by decreasing liver ROS and increasing SOD activity.WSBC pills can increase the protein expression level of SOD1 and NQO-1 to prevent oxidation.WSBC pills can protect the liver by regulating intestinal flora.In chronic ALD model,WSBC pills can increase the activity and expression of ADH in ALD mice,improve the pathological state of liver,and reduce the accumulation of liver fat.WSBC pills can reduce the expressions of FASN,SREBP1 and CD36,and reduce the expression of IL-la gene in ALD mice.After antibiotic intervention,the liver protection and lipid-lowering effects of WSBC pills decrease.WSBC pills affect the species composition and relative abundance of intestinal microflora in ALD mice.WSBC pills could significantly increase the abundance of Bifidobacterium,and decrease the abundance of Alloprevotella、Mucispirillum and Olsenella.Conclusion and SignificanceWSBC pills can alleviate the intoxication symptoms of mice by improving alcohol metabolism and protect mice from acute alcoholic liver injury by anti-oxidative stress.WSBC pills can reduce the accumulation of liver lipids by reducing the generation and transport of lipids to prevent chronic ALD,and has a certain anti-inflammatory effect.At the same time,WSBC pills also play a positive role in the prevention of ALD by regulating the intestinal flora of mice.We innovatively use WSBC pills to prevent ALD,and explore its pharmacodynamics and mechanisms.We find a new idea for the clinical application of WSBC pills,and find a new way to prevent and treat ALD.