Effects of Duhuo Jisheng Decoction on intestinal flora of rats with knee osteoarthritis based on 16S rDNA sequencing
Objective To explore the potential mechanism of Duhuo Jisheng Decoction(DHJSD)in regulating intestinal flora for the treatment of knee osteoarthritis(KOA)through 16S rDNA sequencing Methods Eighteen SD rats were randomized into model group(intra-articularly injected with 50 μL of sodium iodoacetate at a concentration of 40 mg/mL),blank group(gavaged with an equal volume of normal saline),and DHJSD group(modeled using the same method as the model group and gavaged with DHJSD at a dose of 16.5 g/kg),with six rats in each group.Intervention began two weeks after modeling,with once-daily gavage for four weeks,during which the rat general condition was observed.After the administration period,samples were collected.Cartilage changes were observed using HE staining.ELISA was used to measure serum levels of inflammatory factors including interleukin(IL)-1β,IL-6,IL-17,and tumor necrosis factor-α(TNF-α).RT-qPCR was employed to determine the relative expression levels of nuclear factor-KB p65(NF-κB p65),mechanistic target of rapamycin(mTOR),AMP-activated protein kinase(AMPK),and phospha-tidylinositol 3-kinase(PI3K)in the synovium.Fecal samples were collected for 16S rDNA sequencing to analyze differences in the intestinal flora.Results Compared with the blank group,rats in the model group exhibited decreased activity,lusterless fur,and reduced food and water intake.In contrast,the DHJSD group showed significant improvements in activity,fur luster,and food intake compared to the model group.The HE staining indicated that DHJSD effectively reduced cartilage damage,fibrosis,and inflammatory responses.The ELISA showed that,compared with the blank group,the levels of IL-1β,IL-6,IL-17,and TNF-α in the model group were significantly higher(P＜0.01).However,compared with the model group,the levels of 11-1β and TNF-α in the DHJSD group were significantly lower(P＜0.01).The RT-qPCR indicated that,compared with the blank group,the relative expression levels of NF-κB p65,mTOR,and AMPK significantly increased in the model group(P＜0.05).Compared with the model group,the relative expression level of mTOR in the DHJSD group significantly decreased(P＜0.05).The 16S rDNA sequencing showed that DHJSD regulated the intestinal microbiota structure in KOA rats,and affected both α and β diversity of the microbiota.Lactococcus and Enterorhabdus were the most significantly enriched genera in the DHJSD and model groups,respectively.Conclusion DHJSD can effectively repair cartilage damage and slow down the progression of KOA.Its mechanism may be related to regulating intestinal flora,maintaining intestinal barrier integrity,and reducing immune-inflammatory responses.
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