Background:This research aims to investigate potential gene targets and mechanisms through which the traditional Chinese medicine(TCM)Bupleuri Radix(Chaihu)may impact on clear cell renal cell carcinoma(ccRCC)treatment...Background:This research aims to investigate potential gene targets and mechanisms through which the traditional Chinese medicine(TCM)Bupleuri Radix(Chaihu)may impact on clear cell renal cell carcinoma(ccRCC)treatment.Methods:Public databases were employed to identify Cedrenol,an active component of Bupleuri Radix,and its associated gene targets.Among these,the gene RORC(also known as RORγ)was selected for detailed analysis due to its high expression correlation.Expression levels of RORC across various cancers were assessed using the TCGA pan-cancer dataset.Further analyses,including differential expression,prognostic relevance,immune infiltration,single-gene,and functional enrichment analyses,were conducted using the TCGA ccRCC dataset.Additionally,potential drug sensitivities and molecular docking interactions with RORC were explored using the GSCALite and CellMiner databases.The effects of RORC on ccRCC were also validated through cellular experiments.Results:RORC exhibited elevated expression in clear cell renal carcinoma tissue in contrast to normal tissues,and lower RORC expression was related to better prognosis.Immune cell infiltration analysis suggested that RORC may influence the penetration of cells that inhibit immune responses,such as regulatory T cells,thereby affecting ccRCC progression.Furthermore,molecular docking studies revealed that several drugs,including Axitinib,Docetaxel,Methotrexate,and Temsirolimus,have a high affinity for RORC and exhibit strong molecular binding.In cellular experiment experiments confirmed that RORC knockdown led to reduced proliferation,metastasis,and ccRCC cell invasion.Conclusion:The study implies that RORC may be a potential gene target for Bupleuri Radix in the management of ccRCC.展开更多
目的:探讨四妙汤改善类风湿关节炎(RA)模型大鼠的作用机制。方法:采用胶原诱导法(CIA)建立RA模型大鼠,灌胃四妙汤水煎液低、中、高剂量组(3.15 g/kg、6.3 g/kg、12.6 g/kg),维生素D组(25 μg/kg)。给药周期为28天,记录大鼠一般情况。苏...目的:探讨四妙汤改善类风湿关节炎(RA)模型大鼠的作用机制。方法:采用胶原诱导法(CIA)建立RA模型大鼠,灌胃四妙汤水煎液低、中、高剂量组(3.15 g/kg、6.3 g/kg、12.6 g/kg),维生素D组(25 μg/kg)。给药周期为28天,记录大鼠一般情况。苏木精–伊红(HE)染色法观察踝关节组织病理改变;酶联免疫吸附剂检测法(ELISA)检测大鼠血清白细胞介素-17 (IL-17)、白细胞介素-10 (IL-10)、转化生长因子-β (TGF-β)含量;实时荧光定量PCR法(RT-PCR)检测大鼠关节滑膜组织中维生素D受体(VDR)、1-α羟化酶(CYP27B1)、24-羟化酶(CYP24A1)、叉头/翼状螺旋转录因子3 (Foxp3)、维甲酸相关孤儿核受体γt (RORγt) mRNA表达。结果:与空白组相比,模型组血清IL-17含量水平显著上升(P 0.05),血清IL-10、TGF-β含量水平显著下降(P 0.05);关节滑膜CYP24A1、RORγt mRNA表达水平显著上升(P 0.05),VDR、CYP27B1、Foxp3 mRNA表达水平显著下降(P 0.05)。与模型组相比,维生素D组及四妙汤高剂量组血清IL-17水平显著下降(P 0.05),维生素D组及四妙汤中、高剂量组血清IL-10水平显著上升(P 0.05),维生素D组及四妙汤高剂量组血清TGF-β水平显著上升(P 0.05);维生素D组及四妙汤中、高剂量组关节滑膜VDR、CYP27B1、Foxp3 mRNA表达水平显著上升(P 0.05),维生素D组及四妙汤低、中、高剂量组关节滑膜CYP24A1 mRNA表达水平显著下降(P 0.05),维生素D组及四妙汤中、高剂量组关节滑膜RORγt mRNA表达水平显著下降(P 0.05)。结论:四妙汤通过调节关节滑膜组织中VDR、CYP27B1、CYP24A1、Foxp3、RORγt mRNA表达情况,改善血清IL-17、IL-10、TGF-β水平,发挥其干预RA的作用。Objective: To explore the mechanism of action of Simiao decoction in improving the rat model of RA. Methods: Rat models of rheumatoid arthritis (RA) were established by collagen-induced arthritis (CIA). The rats were then administered with low, medium and high doses of Simiao decoction (3.15 g/kg, 6.3 g/kg, 12.6 g/kg) and vitamin D (25 ng/kg) by gavage. The administration period was 28 days, and the general conditions of the rats were recorded. Observation of pathological changes in ankle joint tissues by hematoxylin-eosin (HE) staining method. The contents of interleukin-17 (IL-17), interleukin-10 (IL-10), and transforming growth factor-β (TGF-β) in the serum of rats were detected by enzyme-linked immunosorbent assay (ELISA). The mRNA expressions of VDR, CYP27B1, CYP24A1, Foxp3, and RORγt in the synovial tissue of rat joints were detected by real-time fluorescence quantitative PCR (RT-PCR). Results: Compared with the control group, the serum IL-17 content level in the model group significantly increased (P 0.05), while the serum IL-10 and TGF-β content levels significantly decreased (P 0.05);the mRNA expression levels of CYP24A1 and RORγt in the synovial membrane significantly increased (P 0.05), and the mRNA expression levels of VDR, CYP27B1, and Foxp3 significantly decreased (P 0.05). Compared with the model group, the serum IL-17 level in the vitamin D group and the high-dose Simiao decoction group significantly decreased (P 0.05), the serum IL-10 level in the vitamin D group and the medium and high-dose Simiao decoction groups significantly increased (P 0.05), and the serum TGF-β level in the vitamin D group and the high-dose Simiao decoction group significantly increased (P 0.05);the mRNA expression levels of VDR, CYP27B1, and Foxp3 in the synovial membrane in the vitamin D group and the medium and high-dose Simiao decoction groups significantly increased (P 0.05), the mRNA expression level of CYP24A1 in the synovial membrane in the vitamin D group and the low, medium, and high-dose Simiao decoction groups significantly decreased (P 0.05), and the mRNA expression level of RORγt in the synovial membrane in the vitamin D group and the medium- and high-dose Simiao decoction groups significantly decreased (P 0.05). Conclusion: Simiao decoction exerts its intervention effect on RA by regulating the mRNA expression of VDR, CYP27B1, CYP24A1, Foxp3, and RORγt in the synovial membrane and improving the levels of serum IL-17, IL-10, and TGF-β.展开更多
The intestinal immune system maintains tolerance to harmless food proteins and gut microbiota through peripherally derived RORγt+Tregs(pTregs),which prevent food intolerance and inflammatory bowel disease.Recent stud...The intestinal immune system maintains tolerance to harmless food proteins and gut microbiota through peripherally derived RORγt+Tregs(pTregs),which prevent food intolerance and inflammatory bowel disease.Recent studies suggested that RORγt+antigen-presenting cells(APCs),which encompass rare dendritic cell(DC)subsets and type 3 innate lymphoid cells(ILC3s),are key to pTreg induction.Here,we developed a mouse with reduced RORγt+APCs by deleting a specific cis-regulatory element of Rorc encoding RORγt.Single-cell RNA sequencing and flow cytometry analyses confirmed the depletion of a RORγt+DC subset and ILC3s.These mice showed a secondary reduction in pTregs,impaired tolerance to oral antigens,and an increase in T helper(Th)2 cells.Conversely,ILC3-deficient mice showed no pTregs or Th2 cell abnormalities.Lineage tracing revealed that RORγt+DCs share a lymphoid origin with ILC3s,consistent with their similar phenotypic traits.These findings highlight the role of lymphoid RORγt+DCs in maintaining intestinal immune balance and preventing conditions like food allergies.展开更多
基金supported by Hubei Provincial Department of Science and Technology,Innovation and Development Joint Fund(No.2022CFD145)Young Talents Project of Hubei Provincial Administration of Traditional Chinese Medicine(No.ZY2023Q002).
文摘Background:This research aims to investigate potential gene targets and mechanisms through which the traditional Chinese medicine(TCM)Bupleuri Radix(Chaihu)may impact on clear cell renal cell carcinoma(ccRCC)treatment.Methods:Public databases were employed to identify Cedrenol,an active component of Bupleuri Radix,and its associated gene targets.Among these,the gene RORC(also known as RORγ)was selected for detailed analysis due to its high expression correlation.Expression levels of RORC across various cancers were assessed using the TCGA pan-cancer dataset.Further analyses,including differential expression,prognostic relevance,immune infiltration,single-gene,and functional enrichment analyses,were conducted using the TCGA ccRCC dataset.Additionally,potential drug sensitivities and molecular docking interactions with RORC were explored using the GSCALite and CellMiner databases.The effects of RORC on ccRCC were also validated through cellular experiments.Results:RORC exhibited elevated expression in clear cell renal carcinoma tissue in contrast to normal tissues,and lower RORC expression was related to better prognosis.Immune cell infiltration analysis suggested that RORC may influence the penetration of cells that inhibit immune responses,such as regulatory T cells,thereby affecting ccRCC progression.Furthermore,molecular docking studies revealed that several drugs,including Axitinib,Docetaxel,Methotrexate,and Temsirolimus,have a high affinity for RORC and exhibit strong molecular binding.In cellular experiment experiments confirmed that RORC knockdown led to reduced proliferation,metastasis,and ccRCC cell invasion.Conclusion:The study implies that RORC may be a potential gene target for Bupleuri Radix in the management of ccRCC.
文摘目的:探讨四妙汤改善类风湿关节炎(RA)模型大鼠的作用机制。方法:采用胶原诱导法(CIA)建立RA模型大鼠,灌胃四妙汤水煎液低、中、高剂量组(3.15 g/kg、6.3 g/kg、12.6 g/kg),维生素D组(25 μg/kg)。给药周期为28天,记录大鼠一般情况。苏木精–伊红(HE)染色法观察踝关节组织病理改变;酶联免疫吸附剂检测法(ELISA)检测大鼠血清白细胞介素-17 (IL-17)、白细胞介素-10 (IL-10)、转化生长因子-β (TGF-β)含量;实时荧光定量PCR法(RT-PCR)检测大鼠关节滑膜组织中维生素D受体(VDR)、1-α羟化酶(CYP27B1)、24-羟化酶(CYP24A1)、叉头/翼状螺旋转录因子3 (Foxp3)、维甲酸相关孤儿核受体γt (RORγt) mRNA表达。结果:与空白组相比,模型组血清IL-17含量水平显著上升(P 0.05),血清IL-10、TGF-β含量水平显著下降(P 0.05);关节滑膜CYP24A1、RORγt mRNA表达水平显著上升(P 0.05),VDR、CYP27B1、Foxp3 mRNA表达水平显著下降(P 0.05)。与模型组相比,维生素D组及四妙汤高剂量组血清IL-17水平显著下降(P 0.05),维生素D组及四妙汤中、高剂量组血清IL-10水平显著上升(P 0.05),维生素D组及四妙汤高剂量组血清TGF-β水平显著上升(P 0.05);维生素D组及四妙汤中、高剂量组关节滑膜VDR、CYP27B1、Foxp3 mRNA表达水平显著上升(P 0.05),维生素D组及四妙汤低、中、高剂量组关节滑膜CYP24A1 mRNA表达水平显著下降(P 0.05),维生素D组及四妙汤中、高剂量组关节滑膜RORγt mRNA表达水平显著下降(P 0.05)。结论:四妙汤通过调节关节滑膜组织中VDR、CYP27B1、CYP24A1、Foxp3、RORγt mRNA表达情况,改善血清IL-17、IL-10、TGF-β水平,发挥其干预RA的作用。Objective: To explore the mechanism of action of Simiao decoction in improving the rat model of RA. Methods: Rat models of rheumatoid arthritis (RA) were established by collagen-induced arthritis (CIA). The rats were then administered with low, medium and high doses of Simiao decoction (3.15 g/kg, 6.3 g/kg, 12.6 g/kg) and vitamin D (25 ng/kg) by gavage. The administration period was 28 days, and the general conditions of the rats were recorded. Observation of pathological changes in ankle joint tissues by hematoxylin-eosin (HE) staining method. The contents of interleukin-17 (IL-17), interleukin-10 (IL-10), and transforming growth factor-β (TGF-β) in the serum of rats were detected by enzyme-linked immunosorbent assay (ELISA). The mRNA expressions of VDR, CYP27B1, CYP24A1, Foxp3, and RORγt in the synovial tissue of rat joints were detected by real-time fluorescence quantitative PCR (RT-PCR). Results: Compared with the control group, the serum IL-17 content level in the model group significantly increased (P 0.05), while the serum IL-10 and TGF-β content levels significantly decreased (P 0.05);the mRNA expression levels of CYP24A1 and RORγt in the synovial membrane significantly increased (P 0.05), and the mRNA expression levels of VDR, CYP27B1, and Foxp3 significantly decreased (P 0.05). Compared with the model group, the serum IL-17 level in the vitamin D group and the high-dose Simiao decoction group significantly decreased (P 0.05), the serum IL-10 level in the vitamin D group and the medium and high-dose Simiao decoction groups significantly increased (P 0.05), and the serum TGF-β level in the vitamin D group and the high-dose Simiao decoction group significantly increased (P 0.05);the mRNA expression levels of VDR, CYP27B1, and Foxp3 in the synovial membrane in the vitamin D group and the medium and high-dose Simiao decoction groups significantly increased (P 0.05), the mRNA expression level of CYP24A1 in the synovial membrane in the vitamin D group and the low, medium, and high-dose Simiao decoction groups significantly decreased (P 0.05), and the mRNA expression level of RORγt in the synovial membrane in the vitamin D group and the medium- and high-dose Simiao decoction groups significantly decreased (P 0.05). Conclusion: Simiao decoction exerts its intervention effect on RA by regulating the mRNA expression of VDR, CYP27B1, CYP24A1, Foxp3, and RORγt in the synovial membrane and improving the levels of serum IL-17, IL-10, and TGF-β.
文摘The intestinal immune system maintains tolerance to harmless food proteins and gut microbiota through peripherally derived RORγt+Tregs(pTregs),which prevent food intolerance and inflammatory bowel disease.Recent studies suggested that RORγt+antigen-presenting cells(APCs),which encompass rare dendritic cell(DC)subsets and type 3 innate lymphoid cells(ILC3s),are key to pTreg induction.Here,we developed a mouse with reduced RORγt+APCs by deleting a specific cis-regulatory element of Rorc encoding RORγt.Single-cell RNA sequencing and flow cytometry analyses confirmed the depletion of a RORγt+DC subset and ILC3s.These mice showed a secondary reduction in pTregs,impaired tolerance to oral antigens,and an increase in T helper(Th)2 cells.Conversely,ILC3-deficient mice showed no pTregs or Th2 cell abnormalities.Lineage tracing revealed that RORγt+DCs share a lymphoid origin with ILC3s,consistent with their similar phenotypic traits.These findings highlight the role of lymphoid RORγt+DCs in maintaining intestinal immune balance and preventing conditions like food allergies.
