The modulating of Qingguang'an Ⅱ Formula on gut microbiota in mice with chronic high intraocular pressure by 16S rDNA sequencing
10.1016/j.dcmed.2025.01.006
- VernacularTitle:基于16S rDNA测序探讨青光安Ⅱ号方对慢性高眼压模型小鼠肠道菌群的影响
- Author:
Yasha ZHOU
1
;
Wenyong GAO
;
Yu HUANG
;
Xin XIA
;
Li XIAO
;
Ying DENG
;
Qinghua PENG
;
Jun PENG
Author Information
1. 湖南中医药大学中西医结合学院,湖南 长沙 410208,中国
- Publication Type:Journal Article
- Keywords:
Qingguang'an Ⅱ Formula;
Chronic high intraocular pressure;
16S rDNA sequencing;
Optic neuroprotection;
Gut microbiota
- From:
Digital Chinese Medicine
2024;7(4):332-342
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of Qingguang'an Ⅱ Formula(QGA Ⅱ)on the gut micro-biota of mice with chronic high intraocular pressure(IOP)model,and explore its key micro-biota for protecting the optic nerve.Methods A total of 10 specific pathogen free(SPF)grade female DBA/2J mice were random-ly divided into model group and QGA Ⅱ group(n=5 for each group),while additional 5 SPF-grade female C57BL/6J mice were assigned to control group.Mice presented spontaneous high IOP and showed elevated approximately at the age of seven months.The high IOP was maintained until week 38,when gavage was initiated.Mice in control group underwent the same intragastric treatment,while those in QGA Ⅱ group were gavaged with QGA Ⅱ(9.67 g/kg),once a day for four weeks.Retinal morphology was examined using hematoxylin and eosin(HE)staining,with the number of retinal ganglion cells(RGCs)counted.The expression level of Brn3a protein,a specific marker for RGCs,was detected by immunofluorescence,with the mean optical density(OD)measured for quantitative analysis.In addition,16S rDNA se-quencing was leveraged to analyze changes in the diversity of gut microbiota,including their α-diversity(Chao1,Shannon,Pielou's evenness,and observed species index)and β-diversity.Venn diagrams and linear discriminant analysis effect size(LEfSe)analysis was employed to investigate the number of amplicon sequence variants(ASVs),the abundance of differential gut microbiota species,and the classification of species at both the phylum and genus levels within the three groups of mice.Results HE staining revealed that compared with control group,model group showed signif-icant reduction in the number of RGCs(P<0.01),with intracellular vacuolar degeneration and nuclear pyknosis.After QGA Ⅱ treatment,the number of RGCs was significantly in-creased compared with model group(P<0.01),with notable improvements in intracellular vacuolar degeneration.Immunofluorescence analysis showed that the mean OD of Brn3a protein was significantly decreased in model group compared with control group(P<0.01),while QGA Ⅱ treatment significantly elevated its expression level(P<0.01).Analysis of α-diversity showed that after QGA Ⅱ intervention,the Chao1,Shannon,and Pielou's evenness indices were significantly increased(P<0.01),and the observed species index was elevated(P<0.05).β-Diversity analysis demonstrated distinct clustering among the three groups,indicating relatively low similarity in bacterial community structures.ASV clustering identi-fied a total of 14 061 ASVs across all groups,with 9 514 ASVs shared between model and QGA Ⅱ groups.At the phylum level,the abundance of Bacteroidetes was significantly decreased in model group compared with control group(P<0.01),while Firmicutes and the Firmicutes/Bacteroidetes(F/B)ratio were significantly increased(P<0.01).QGA Ⅱ treatment significantly reduced both Firmicutes abundance and the F/B ratio(P<0.01).At the genus level,Lactobacillus was dominant across all groups,with its abundance significantly in-creased in model group(P<0.01)and subsequently decreased following QGA Ⅱ intervention(P<0.05).Conclusion QGA Ⅱ restructured the gut microbiota of DBA/2J mice with chronic high IOP,bringing changes in their diversity and abundance of components.Firmicutes,Bacteroidetes,Lactobacillus,along with their associated microorganisms,are likely critical components of the gut microbiota that contribute to the optic neuroprotective effects of QGA Ⅱ on chronic high IOP mice.
