BACKGROUND:Developed by the esteemed Chinese medicine master Wei Guikang,Tongan Decoction has proven highly effective in treating knee osteoarthritis.However,the mechanism of action is yet unclear.OBJECTIVE:To elucidate how Tongan Decoction modulates synovial macrophage polarization as a therapeutic strategy for knee osteoarthritis in a rat model.METHODS:(1)We employed high-throughput microRNA sequencing and polymerase chain reaction to analyze the differentially expressed miRNA in synovial macrophages of normal and knee osteoarthritis rats.Predicted target genes of miR-27a were identified using bioinformatics databases,with subsequent validation through luciferase assays.A total of 68 Sprague-Dawley rats were randomly divided into normal control group(n=16),model group(n=16),miR-27a overexpression group(n=12),Tongan Decoction group(n=12),and Tongan Decoction+miR-27a inhibition group(n=12).The miR-27a overexpression group and the Tongan Decoction+miR-27a inhibition group were injected with miR-27a mimic and miR-27a inhibitor in the right knee joint cavity,respectively,once daily for 5 continuous days.On the 15th day after modeling,the Tongan Decoction group and the Tongan Decoction+miR-27a inhibition group were given Tongan Decoction by gastric lavage,and the other three groups were given saline by gastric lavage,once daily for 14 continuous days.After administration,behavioral tests,X-rays,hematoxylin-eosin staining of synovial and cartilage tissues of the knee joint and immunofluorescence staining of synovial tissues of the knee joint,RT-PCR,and Western blot were performed.RESULTS AND CONCLUSION:High-throughput sequencing of miRNA showed low expression of miR-27a in synovial tissues of rats with knee osteoarthritis.The target gene of miR-27a was nuclear factor κB,and luciferase assay showed that the two could bind to each other.Behavioral assays showed that miR-27a overexpression or Tongan Decoction alleviated joint dysfunction in rats with knee osteoarthritis,and miR-27a inhibition antagonized the effect of Tongan Decoction.X-ray films and hematoxylin-eosin staining showed that miR-27a overexpression or Tongan Decoction reduced the degree of knee osteoarthritis and miR-27a inhibition weakened the therapeutic effect of Tongan Decoction.The results of RT-PCR and western blot assay showed that compared with the normal control group,the expression of interleukin 10 was reduced in the model group(P＜0.05),and the expression of matrix metalloproteinase 13,interleukin 1β,and nuclear factor κB was elevated in the model group(P＜0.05).miR-27a overexpression or Tongan Decoction could differently reverse the changes in the above-mentioned indexes,while miR-27a inhibition weakened the effect of Tongan Decoction.Immunofluorescence staining results showed that CD86 protein expression in the model group was higher than that in the normal control group(P＜0.05),and CD206 protein expression was lower than that in the normal control group(P＜0.05);miR-27a overexpression group and Tongan Decoction had lower CD86 protein expression than that in the model group(P＜0.05),and higher CD206 protein expression than that in the model group(P＜0.05);in the Tongan Decoction+miR-27a inhibition group,CD86 protein expression was higher(P＜0.05)and CD206 protein expression was lower than that in the Tongan Decoction group(P＜0.05).Tongan Decoction mitigates knee osteoarthritis by upregulating miR-27a expression and suppressing nuclear factor κB activity,which improves knee joint function and further treats knee osteoarthritis.

BACKGROUND:Developed by the esteemed Chinese medicine master Wei Guikang,Tongan Decoction has proven highly effective in treating knee osteoarthritis.However,the mechanism of action is yet unclear.OBJECTIVE:To elucidate how Tongan Decoction modulates synovial macrophage polarization as a therapeutic strategy for knee osteoarthritis in a rat model.METHODS:(1)We employed high-throughput microRNA sequencing and polymerase chain reaction to analyze the differentially expressed miRNA in synovial macrophages of normal and knee osteoarthritis rats.Predicted target genes of miR-27a were identified using bioinformatics databases,with subsequent validation through luciferase assays.A total of 68 Sprague-Dawley rats were randomly divided into normal control group(n=16),model group(n=16),miR-27a overexpression group(n=12),Tongan Decoction group(n=12),and Tongan Decoction+miR-27a inhibition group(n=12).The miR-27a overexpression group and the Tongan Decoction+miR-27a inhibition group were injected with miR-27a mimic and miR-27a inhibitor in the right knee joint cavity,respectively,once daily for 5 continuous days.On the 15th day after modeling,the Tongan Decoction group and the Tongan Decoction+miR-27a inhibition group were given Tongan Decoction by gastric lavage,and the other three groups were given saline by gastric lavage,once daily for 14 continuous days.After administration,behavioral tests,X-rays,hematoxylin-eosin staining of synovial and cartilage tissues of the knee joint and immunofluorescence staining of synovial tissues of the knee joint,RT-PCR,and Western blot were performed.RESULTS AND CONCLUSION:High-throughput sequencing of miRNA showed low expression of miR-27a in synovial tissues of rats with knee osteoarthritis.The target gene of miR-27a was nuclear factor κB,and luciferase assay showed that the two could bind to each other.Behavioral assays showed that miR-27a overexpression or Tongan Decoction alleviated joint dysfunction in rats with knee osteoarthritis,and miR-27a inhibition antagonized the effect of Tongan Decoction.X-ray films and hematoxylin-eosin staining showed that miR-27a overexpression or Tongan Decoction reduced the degree of knee osteoarthritis and miR-27a inhibition weakened the therapeutic effect of Tongan Decoction.The results of RT-PCR and western blot assay showed that compared with the normal control group,the expression of interleukin 10 was reduced in the model group(P＜0.05),and the expression of matrix metalloproteinase 13,interleukin 1β,and nuclear factor κB was elevated in the model group(P＜0.05).miR-27a overexpression or Tongan Decoction could differently reverse the changes in the above-mentioned indexes,while miR-27a inhibition weakened the effect of Tongan Decoction.Immunofluorescence staining results showed that CD86 protein expression in the model group was higher than that in the normal control group(P＜0.05),and CD206 protein expression was lower than that in the normal control group(P＜0.05);miR-27a overexpression group and Tongan Decoction had lower CD86 protein expression than that in the model group(P＜0.05),and higher CD206 protein expression than that in the model group(P＜0.05);in the Tongan Decoction+miR-27a inhibition group,CD86 protein expression was higher(P＜0.05)and CD206 protein expression was lower than that in the Tongan Decoction group(P＜0.05).Tongan Decoction mitigates knee osteoarthritis by upregulating miR-27a expression and suppressing nuclear factor κB activity,which improves knee joint function and further treats knee osteoarthritis.

In recent years, advancements in microfabrication technology and tissue engineering have propelled the development of a novel platform known as organoid-on-a-chip for drug screening and disease modeling. This platform integrates organoids and organ-on-a-chip technologies, emerging as a promising approach for in vitro modeling of human organs. Organ-on-a-chip leverages microfluidic device to simulate the physiological environment of specific organs, offering a more dynamic and flexible setting that can mimic a more comprehensive human biological context. However, the lack of functional vasculature has remained a major challenge in this technology. Vascularization is crucial for the long-term cultivation and in vitro modeling of organoids, which is of great significance in drug development and personalized medical approaches. The authors reviewed the research progress in the construction of vascularized organoid-on-a-chip including the methods for constructing in vitro vascularized models, vascularization of organoids, etc, which may serve as a reference for the construction of fully functional vascularized organoid-on-a-chip.

Objective:To investigate the effects of Bushen Jianpi formula(BSJP)in mediating the inhibitory effects of farnesoid X receptor(FXR)on cancer cachexia(CC)induced by hepatocellular carcinoma AH-130 cells in a rat model and its underlying mechanism.Methods:A liver cancer cachexia rat model was established by intraperitoneal injection of AH-130 cells.The rats were divided into five groups:blank control,model control,CDCA(FXR agonist),BSJP,and BSJP+CDCA groups.After modelling,the rats were treated with CDCA,BSJP,or their combination for consecutive 16 days,and their body weights were measured weekly.At the end of the experiment,the rats were sacrificed,and abdominal aortic blood,feces,and brown adipose tissues from the epididymal,inguinal,and scapular regions were collected.Liquid chromatography-mass spectrometry(LC-MS)was used to detect the composition and content of bile acids in serum and feces of rats.WB and qPCR were used to detect the expression of FXR,Wnt family member 10b(Wnt10b),β-catenin,and uncoupling protein 1(UCP-1)in the ileum,brown adipose,and white adipose tissues of rats.Results:Compared with the blank control group,the body mass of the rats in the model group was significantly reduced(P<0.01);compared with the model control group,the body mass of the rats in CDCA,BSJP and BSJP+CDCA groups all increased(P<0.01).Compared with the blank control group,the epididymal,inguinal,scapular,and total brown adipose tissue mass were all elevated in the model control group(all P<0.05);compared with the model control group,the brown fat mass in the inguinal and epididymis regions of the rats decreased significantly in all treatment groups(P<0.05),but this decrease was not significant in the scapular region(P>0.05);besides,the total brown fat mass decreased notably in all treatment groups compared to the model control group(all P<0.01).LC-MS analysis showed that the composition and content of bile acids in the serum and feces of rats were altered in all groups.qPCR and WB results confirmed that,compared to the model group,BSJP and BSJP+CDCA promoted the mRNA and protein expression of FXR in the ileum,brown adipose,and white adipose tissues of rats(all P<0.05),and decreased the mRNA and protein expression of Wnt10b,β-catenin,UCP-1(all P<0.05).Conclusion:The BSJP formula can inhibit hepatocellular carcinoma cell AH-130-induced cachexia in rats,alleviating the associated body weight loss and brown adipose tissue formation.The mechanism may involve the regulation of FXR and the inhibition of the expression of Wnt10b,β-catenin,and UCP-1 in brown adipose tissue through the Wnt signaling pathway.