Study On The Mechanism Of COSM In The Treatment Of NAFLD Based On The Intestine-liver Axis And The Preparation Of Enteric-coated Dripping Pills

Non-alcoholic fatty liver disease(NAFLD)refers to the pathological state of the fatty liver caused by non-excessive alcohol intake.The harm that NAFLD brings to humans cannot be ignored.At present,there are fewer drugs to treat NAFLD in the market,and there are many shortcomings in applications and poor therapeutic effects.Therefore,it is urgent to find NAFLD therapeutic drugs that are safe,effective,and have few side effects.Chitosan Oligosaccharides(COS)is a marine oligosaccharide composed of glucosamine and a small amount of N-acetylglucosamine connected by β-1,4-glycosidic bonds.It has a variety of biological activities and can regulate the intestinal microbiota ability.Enteric-coated dripping pills have the characteristics of high bioavailability,strong drug stability,and the ability to control drug release in the intestinal tract.Therefore,this article investigated the therapeutic activity of COSM(Average molecular weight≤3000)in the treatment of NAFLD,clarified its mechanism,and prepared COSM as entericcoated dripping pills,and discussed its preparation method and quality inspection.The results of animal experiments show that COSM can significantly improve the abnormal blood glucose level of NAFLD mice and reduce the high insulin level without affecting appetite,thereby improving the overall metabolic disorder.COSM can significantly improve the liver function damage of NAFLD mice and decrease serum AST and ALT levels.COSM can increase the CAT of NAFLD mice,enhance the T-AOC ability,and thus enhance the antioxidant capacity;significantly reduce the serum TNF-α level,increase the serum IL-10 level,and have a certain anti-inflammatory ability.The results of liver lipid determination and pathological analysis show that COSM can significantly improve liver lipid metabolism and reduce liver steatosis.At the same time,COSM can improve the barrier dysfunction of the mouse ileum and colon to a certain extent.The experimental results of the mechanism showed that compared with the Model group mice,the COSM group’s fecal bacterial community abundance and diversity were reduced,which could significantly increase the Akkermansia genus bacteria(P<0.05)and reduce the F/B(Firmicutes/Bacteroidete)ratio,significantly increase the Bacteroides Uniformi(P=0.001)The relative abundance of beneficial bacteria such as Bacteroides acidifaciens(P=0.026)and Clostridium paraputrificum(P=0.020),down-regulated the relative abundance of pathogenic bacteria such as Desulfovibrio sp ABHU2SB(P=0.044)and Lachnospiraceae Bacterium(P=0.020).COSM can reduce serum LPS levels in NAFLD mice.Western Blotting results show that COSM can significantly upregulate colonic tight junction proteins ZO-1 and Occludin.Colon immunofluorescence staining also proves that COSM can upregulate colonic ZO-1 protein expression and improve liver LPS/TLR4/NF-κB pathway,thereby improving the liver disease state of NAFLD mice.The results of the formulation experiment show that the optimal preparation process conditions for COSM drop pill cores are as follows: drug:matrix=1:3,the melting temperature of the drug solution is 80℃,the drip distance is 15 cm,the selected matrix is PEG6000,the condensation medium is liquid paraffin,and the condensation temperature is 5-10°C.According to the process conditions,the drop pill was prepared with round appearance and uniform color.Based on this pellet core,the best process to obtain enteric coating is as follows: 1)Coating liquid: 5% by mass enteric coating material,5% by mass talc,0.5% by mass PEG6000.2)Coating conditions: the inlet air temperature is 40-50℃,the exhaust air temperature is 25-30℃,the material temperature is 25-30℃;the speed of the coating machine is 25 rpm,the atomization pressure is 0.12 Mpa,and the coating is increased.The weight is20%.The enteric-coated dripping pill is white to light gray like round spherical pill without special odor.The properties,weight,and dissolution time limit were checked,and they all met the pharmacopoeia regulations.Each pill contains14.05 mg COSM on average,which meets the set requirements.Successfully established a stable preparation process and quality standards for COSM enteric-coated dripping pills.