Exploring The Mechanism Of Gandouling Intervention For Cognitive Impairment In Wilson’s Disease Based On Intestinal Flora And Its Key Product SCFAs

Objective: Using 16 S r RNA technology and gas chromatography,we observed the changes of intestinal flora and its metabolite short-chain fatty acids in a rat of copper-loaded cognitive impairment,and explored the metabolic mechanisms of intestinal flora and its key product short-chain fatty acids in the treatment of cognitive impairment in Wilson’s disease with Gandouling.Methods: The rats were randomly divided into blank control group,model group,Gandouling group and Penicillamine group,and the blank control group was given normal chow,while Model group,Gandouling group and Penicillamine group was given copper-containing(1g/kg)chow and copper-containing(0.185%)water to produce a copper-loaded rat model.At week 7,the Gandouling group was given Gandouling 0.486g/(kg-d)and the Penicillamine group was given Penicillamine0.09g/(kg-d)by gavage once a day;the blank control group and the model group were given equal amounts of saline for 6 weeks.Moriss water maze experiment at the end of week 12 dosing.Moriss water maze experiment was performed to test the spatial learning and memory ability of rats.16 S r RNA was used to detect the diversity and structural distribution of fecal flora in the cecum of rats,and gas chromatography was used to quantify the content of short-chain fatty acids in the cecum of rats.Results:(1)Six weeks after modeling of copper-loaded rats,the escape latency of rats in the model group,Gandouling group and Penicillamine group increased significantly,and the number of time crossing platforms and total distance traveled in the platform quadrant decreased significantly(P < 0.05)compared with the blank control rats.After six weeks of administration,compared with the pre-drug period,the escape latency of rats in the model group increased significantly,and the number of platform crossings and total distance traveled in the platform quadrant decreased significantly(P < 0.05),while the escape latency of rats in the Gandouling and Penicillamine groups decreased significantly,and the number of platform crossings and total distance traveled in the platform quadrant increased significantly(P < 0.05).1 At the class level,the relative abundance of Actinobacteria significantly increased(P < 0.05)and the relative abundance of Firmicutes and Planctomycetes significantly decreased(P < 0.05)in the model group compared with the blank control group;after Gandouling treatment by gavage,the Gandouling group The relative abundance of Firmicutes and Planctomycetes increased significantly(P < 0.05)and the relative abundance of Actinobacteria decreased significantly(P < 0.05)after Gandouling gavage treatment.(2)At the family level,compared with the blank control group,the relative abundance of Enterococcaceae,Eubacteriaceae,Bifidobacteriaceae,and Sutterellaceae increased significantly in the model group(P < 0.05),and the relative abundance of Streptococcaceae,Christensenellaceae,Porphyromonadaceae,and Planctomycetaceae significantly decreased(P < 0.05);after Gandouling gavage treatment,Enterococcaceae,Eubacteriaceae,and Bifidobacteriaceae in the Gandouling group were significantly decreased(P < 0.05).Bifidobacteriaceae,and Sutterellaceae significantly decreased(P < 0.05),and Streptococcaceae,Christensenellaceae,Porphyromonadaceae,and Planctomycetaceae significantly increased after Gandouling gavage treatment(P < 0.05).(3)At the genus level,Enterococcus,Eubacterium,Bifidobacterium,Parvibacter,Olsenella,and Parasutterella were significantly higher in the model group compared to the blank control group(P<0.05);Christensenella,Alloprevotella,Desulfovibrio,Acetatifactor,Murimonas,Strepto coccus,Subdivision5＿genera＿incertae＿sedis,Rombo-utsia,and Bilophila were significantly,reduced(P < 0.05);after Gandouling gavage treatment,Enterococcus,Eubacterium,Bifidobacterium,and Parvibacter,Olsenella,and Parasutterella were significantly reduced after Gandouling gavage(P < 0.05),and Christensenella,Alloprevotella,Desulfovibrio,Acetatifactor,Murimonas,and Streptococcus,Subdivision5＿genera＿incertae＿sedis,Romboutsia,and Bilophila were significantly increased(P < 0.05).(3)The content of acetic acid,butyric acid,propionic acid,and valeric acid was significantly decreased in the model group compared with in the blank group(VIP>1,P<0.05),and the content of acetic acid,butyric acid,propionic acid,and valeric acid was significantly increased in the Gandouling group after Gandouling intervention compared with in the model group(VIP>1,P<0.05).(4)Correlation analysis between short-chain fatty acids and intestinal flora: Acetic acid was positively correlated with Desulfovibrio,Murimonas,Streptococcus,and Romboutsia spp.and negatively correlated with Parasutterella,Bifidobacterium,and Parvibacter.Propionic acid was positively correlated with Murimonas,Streptococcus,Romboutsia,and negatively correlated with Parasutterella,Bifidobacterium,and Parvibacter.Butyric acid was positively correlated with Desulfovibrio,Romboutsia and negatively correlated with Parasutterella.Valeric acid was positively correlated with Desulfovibrio,Murimonas,Streptococcus,Romboutsia,and negatively correlated with Parasutterella,Bifidobacterium,Parvibacter.Conclusions: Gandouling improves cognitive function in a copper-loaded rat,and the mechanism may be through intestinal flora to regulate the metabolism of short-chain fatty acids,thereby improving cognitive impairment in copper-loaded rats.