Effects Of Artemisia Annua Turtle Carapace On Intestinal Microorganisms In MRL/lpr Lupus Mice

Objective To investigate the effects of Artemisia annua,Turtle Carapace and Artemisia annua-Turtle Carapace on MRL/lpr lupus mice,and to elucidate the effects of Artemisia annua Turtle Carapace on intestinal microorganisms in MR L/lpr lupus mice from the perspective of intestinal microorganisms.Methods 40 SPF grade female MRL/lpr lupus mice were randomly divided into model group,hormone group,low dose Artemisia annua group(0.12 g/m L),high dose Artemisia annua group(0.24 g/m L),low dose Turtle Carapace group(0.3 g/m L),high dose Turtle Carapace group(0.6 g/m L),low dose Artemisia annua Turtle Carapace group(0.42 g/m L),high dose Artemisia annua Turtle Carapace group(0.84 g/m L).After 10 weeks,mice were executed and blood,feces and intestinal tissues were collected.The serum creatinine(CREA)and urea nitrogen(BUN)in lupus mice were detected by automatic blood biochemical analyzer;the serum levels of tumor necrosis factor(TNF-α)and urinary protein were detected by ELISA;the contents of TLR-4,TLR-9,IL-1beta and IFN-α in intestinal tissues of lupus mice were detected by RT-q PCR;and the structure of fecal intestinal microorganisms in lupus mice was analyzed by Illumina Miq high throughput sequencing technology.The correlation between different microorganisms and different microflora was analyzed by Spearman correlation analysis.Result 1.Indicators and urinary protein results of each group: Compared with model group mice,serum CREA,BUN and TNF-α in hormone group and Artemisia annua Turtle Carapace group were significantly lower(P<0.05,P<0.01),among which low dose Artemisia turtle group,high dose Turtle Carapace group,Artemisia annua Turtle Carapace group had better effect;compared with model group,Urinary protein was significantly decreased after treatment in hormone group and high dose Artemisia annua Turtle Carapace group(P<0.01).2.Detection of cytokines in intestinal tissues: Compared with model group,except high-dose Artemisia annua group and low-dose Turtle Carapace group,the expression levels of IFN-α in other groups were significantly decreased(P<0.01);the expression levels of IL-1β and TLR4 in hormone group,low-dose Artemisia annua group,high-dose Turtle Carapace group,low-dose Artemisia annua turtle group,high-dose Artemisia annua turtle group.3.Alpha diversity of intestinal flora in each group: Compared with model mice,the diversity and abundance index of intestinal flora in low-dose Artemisia annua group,high-dose Turtle Carapace group and high-dose Artemisia annua Turtle Carapacel group increased(P<0.05,P<0.01).4.Differential analysis of intestinal microorganisms among different groups: Compared with other groups,Staphylococcus and Lactococcus are the unique differential microbial genera in hormone group;Lactobacillus,Allobaculum,Sutterella,Dehalobacterium,Coprococcus,Dorea,Oscillospira and Desulfovibrio are the unique differential microbial genera in low dose Artemisia annua;[Ruminococcus] is unique in high dose turtle shell group.Bacteroides,Parabacteroides and Coprobacillus are endemic genera of differentiated microorganisms in the high-dose Artemisia sinensis Turtle Carapacel group.5.Spearman correlation analysis of intestinal flora: Clostridium,Bifidobacterium,Coprococcus were positively correlated with TNF-α(P<0.05,P<0.01);Odoribacter,Allobaculum,Sutterella were negatively correlated with TNF-α(P<0.05,P<0.01);Odoribacter,Helicobacter,Parabacteroids,Coprococcus were negatively correlated with CREA(P<0.05,P<0.01).Conclusion 1.Each treatment group of Artemisia annua Turtle Carapace can improve the condition of lupus mice,among which low dose Artemisia annua group,high dose Turtle Carapace group and high dose Artemisia annua Turtle Carapace group have the best effect.Compared with single drug group and drug pair group,the combined effect of Artemisia annua Turtle Carapace group is the best.2.Artemisia annua group,Turtle Carapace group and Artemisia annua Turtle Carapace group can regulate the structure of intestinal microorganisms,and regulate the impact of intestinal microbial species has a certain difference.3.Clostridium,Bifidobacterium,Coprococcus are positively correlated with the condition of MRL/lpr lupus mice,while Odoribacter,Helicobacter,Parabacteroides,Allobaculum,Sutterella,Coprobacillus are negatively correlated with the condition of MRL/lpr lupus mice.