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
Investigate the effects of the classic ancient prescription Danggui Buxue decoction on the vitamin D receptor,25-hydroxyvitamin D-1αhydroxylase,estrogen synthesis aromatase and estrogen receptorβmRNA and protein exp...Investigate the effects of the classic ancient prescription Danggui Buxue decoction on the vitamin D receptor,25-hydroxyvitamin D-1αhydroxylase,estrogen synthesis aromatase and estrogen receptorβmRNA and protein expression in osteoporotic rats.The classic ancient prescription Danggui Buxue decoction may affect the levels of estrogen synthesis aromatase,estrogen receptorβ,FSH,and Klotho in the body through regulating the expression of ovarian vitamin D receptor and 25-hydroxyvitamin D-1αhydroxylase to achieve the therapeutic effect of retinoic acid-induced osteoporosis in rats.The results of this experiment showed that compared with the control group,the serum FSH level of the model group was significantly increased,the Klotho level was decreased,the relative expression of ovarian tissue CYP27B1,CYP19,ERβmRNA and protein was significantly reduced,and the relative expression of vitamin D receptor mRNA and protein was significantly rise.After the intervention of the classic ancient prescription Danggui Buxue decoction and vitamin D,compared with the model group,the serum FSH content decreased,Klotho content increased,ovarian CYP27B1,CYP19,ERβmRNA and protein expression significantly increased,ovarian vitamin D receptor mRNA and protein expression significantly decreased.Based on the effect of the classic ancient prescription Danggui Buxue decoction on the vitamin D system,combined with the results of this experiment,it is concluded that the classic ancient prescription Danggui Buxue decoction may affect the levels of CYP19,ERβ,follicle-stimulating hormone,and Klotho in the body by regulating the expression of ovarian VDR and CYP27B1.The specific mechanism of the treatment of ovarian injury in osteoporosis rats needs further study.Background:Investigate the effects of the classic ancient prescription Danggui Buxue decoction on the vitamin D receptor,25-hydroxyvitamin D-1αhydroxylase,estrogen synthesis aromatase and estrogen receptorβmRNA and protein expression in osteoporotic rats.Methods:The experiment was divided into 6 groups:control group,model group(retinoic acid 100 mg/kg/d),vitamin D group(calcitriol 30 ng/kg/d),the classic ancient prescription Danggui Buxue decoction high,middle and low dose group(7.2 g/kg/d,3.6 g/kg/d,1.8 g/kg/d).Enzyme-linked immunosorbent assay method was used to detect the effects of different doses of the classic ancient prescription Danggui Buxue decoction on serum follicle-stimulating hormone and Klotho levels in retinoic acid-induced osteoporosis rats;real-time fluorescence quantitative PCR was used to detect the classic ancient prescription Danggui Buxue decoction on retinoic acid effect of ovary vitamin D receptor,25-hydroxyvitamin D-1αhydroxylase,estrogen synthesis aromatase,estrogen receptorβmRNA expression in induced osteoporosis rats;Western-blot detection of the classic ancient prescription Danggui Buxue decoction on retinoic acid-induced osteoporosis ovary vitamin D receptor,CYP27B,estrogen synthesis aromatase,estrogen receptorβprotein The impact of expression.Results:Compared with the control group,the serum follicle-stimulating hormone level of the model group was significantly increased,the Klotho level was decreased,the relative expression levels of 25-hydroxyvitamin D-1αhydroxylase,estrogen synthesis aromatase,estrogen receptorβmRNA and protein in the ovarian tissue were significantly decreased,and the relative expression levels of vitamin D receptor mRNA and protein were significantly increased.After the intervention of the classic ancient prescription Danggui Buxue decoction and vitamin D,compared with the model group,the serum follicle-stimulating hormone content decreased,Klotho content increased,ovarian 25-hydroxyvitamin D-1αhydroxylase,estrogen synthesis aromatase,estrogen receptorβmRNA and protein expression significantly increased,ovarian vitamin D receptor mRNA and protein expression significantly decreased.Conclusion:The classic ancient prescription Danggui Buxue decoction may affect the levels of estrogen synthesis aromatase,estrogen receptorβ,follicle-stimulating hormone,and Klotho in the body through regulating the expression of ovarian vitamin D receptor and 25-hydroxyvitamin D-1αhydroxylase to achieve the therapeutic effect of retinoic acid-induced osteoporosis in rats.展开更多
The objective of the study was to determine the role of vitamin D3(VD3) in regulating adaptation and mechanism of rats to low-phosphorus(P) diets. Rats were assigned to 4 diets containing 0.2%, 0.4%, 0.6%,or 0.8% P co...The objective of the study was to determine the role of vitamin D3(VD3) in regulating adaptation and mechanism of rats to low-phosphorus(P) diets. Rats were assigned to 4 diets containing 0.2%, 0.4%, 0.6%,or 0.8% P consisting of 5 replicate cages with 6 rats per replicate cage and fed for 7 days. Four rats from each replicate cage were treated with ethane-1-hydroxy-1,1-diphosphonicacid, tetrasodium salt(EHDP)and 2 rats remained untreated. Twelve hours prior to preparation on d 7, two of the EHDP-treated rats received an intraperitoneal injection of VD3 [1,25-(OH)_2 D_3] at 600 ng per kg body weight, while two rats did not receive the injection. Rats that did not receive VD_3 injection had decreased(P < 0.001) P absorption, but injection of VD3 resulted in increased(P < 0.001) absorption. The effect of VD3 injection was greater(P < 0.001) for rats fed 0.2% P diet than rats fed 0.8% P diet in ileum. Sodium dependent phosphate cotransporter type IIb(Na/Pi-IIb) and 25-hydroxyvitamin D 1-α hydroxylase(CYP27 B1) mRNA level showed the same trend with P absorption. Serum concentration of VD3 and la-hydroxylase activity in rats fed 0.2% P diet were lower than those fed 0.8% P diet. The injection of VD3 increased(P < 0.001)serum concentration of VD3 and la-hydroxylase activity. Thus, VD3 increased Na/Pi-Ⅱb and CYP27 B1 mRNA level and improved serum concentration of VD3 and la-hydroxylase activity in rats fed low-P diets.展开更多
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Investigate the effects of the classic ancient prescription Danggui Buxue decoction on the vitamin D receptor,25-hydroxyvitamin D-1αhydroxylase,estrogen synthesis aromatase and estrogen receptorβmRNA and protein expression in osteoporotic rats.The classic ancient prescription Danggui Buxue decoction may affect the levels of estrogen synthesis aromatase,estrogen receptorβ,FSH,and Klotho in the body through regulating the expression of ovarian vitamin D receptor and 25-hydroxyvitamin D-1αhydroxylase to achieve the therapeutic effect of retinoic acid-induced osteoporosis in rats.The results of this experiment showed that compared with the control group,the serum FSH level of the model group was significantly increased,the Klotho level was decreased,the relative expression of ovarian tissue CYP27B1,CYP19,ERβmRNA and protein was significantly reduced,and the relative expression of vitamin D receptor mRNA and protein was significantly rise.After the intervention of the classic ancient prescription Danggui Buxue decoction and vitamin D,compared with the model group,the serum FSH content decreased,Klotho content increased,ovarian CYP27B1,CYP19,ERβmRNA and protein expression significantly increased,ovarian vitamin D receptor mRNA and protein expression significantly decreased.Based on the effect of the classic ancient prescription Danggui Buxue decoction on the vitamin D system,combined with the results of this experiment,it is concluded that the classic ancient prescription Danggui Buxue decoction may affect the levels of CYP19,ERβ,follicle-stimulating hormone,and Klotho in the body by regulating the expression of ovarian VDR and CYP27B1.The specific mechanism of the treatment of ovarian injury in osteoporosis rats needs further study.Background:Investigate the effects of the classic ancient prescription Danggui Buxue decoction on the vitamin D receptor,25-hydroxyvitamin D-1αhydroxylase,estrogen synthesis aromatase and estrogen receptorβmRNA and protein expression in osteoporotic rats.Methods:The experiment was divided into 6 groups:control group,model group(retinoic acid 100 mg/kg/d),vitamin D group(calcitriol 30 ng/kg/d),the classic ancient prescription Danggui Buxue decoction high,middle and low dose group(7.2 g/kg/d,3.6 g/kg/d,1.8 g/kg/d).Enzyme-linked immunosorbent assay method was used to detect the effects of different doses of the classic ancient prescription Danggui Buxue decoction on serum follicle-stimulating hormone and Klotho levels in retinoic acid-induced osteoporosis rats;real-time fluorescence quantitative PCR was used to detect the classic ancient prescription Danggui Buxue decoction on retinoic acid effect of ovary vitamin D receptor,25-hydroxyvitamin D-1αhydroxylase,estrogen synthesis aromatase,estrogen receptorβmRNA expression in induced osteoporosis rats;Western-blot detection of the classic ancient prescription Danggui Buxue decoction on retinoic acid-induced osteoporosis ovary vitamin D receptor,CYP27B,estrogen synthesis aromatase,estrogen receptorβprotein The impact of expression.Results:Compared with the control group,the serum follicle-stimulating hormone level of the model group was significantly increased,the Klotho level was decreased,the relative expression levels of 25-hydroxyvitamin D-1αhydroxylase,estrogen synthesis aromatase,estrogen receptorβmRNA and protein in the ovarian tissue were significantly decreased,and the relative expression levels of vitamin D receptor mRNA and protein were significantly increased.After the intervention of the classic ancient prescription Danggui Buxue decoction and vitamin D,compared with the model group,the serum follicle-stimulating hormone content decreased,Klotho content increased,ovarian 25-hydroxyvitamin D-1αhydroxylase,estrogen synthesis aromatase,estrogen receptorβmRNA and protein expression significantly increased,ovarian vitamin D receptor mRNA and protein expression significantly decreased.Conclusion:The classic ancient prescription Danggui Buxue decoction may affect the levels of estrogen synthesis aromatase,estrogen receptorβ,follicle-stimulating hormone,and Klotho in the body through regulating the expression of ovarian vitamin D receptor and 25-hydroxyvitamin D-1αhydroxylase to achieve the therapeutic effect of retinoic acid-induced osteoporosis in rats.
基金supported by grants from the Nature Science Foundation (31201810, to M H Cao 31572419, to R J Fang)+1 种基金Education Department of Hunan Province(138039, to M H Cao)Innovation Team Funds Of Hunan Province (to J H He)
文摘The objective of the study was to determine the role of vitamin D3(VD3) in regulating adaptation and mechanism of rats to low-phosphorus(P) diets. Rats were assigned to 4 diets containing 0.2%, 0.4%, 0.6%,or 0.8% P consisting of 5 replicate cages with 6 rats per replicate cage and fed for 7 days. Four rats from each replicate cage were treated with ethane-1-hydroxy-1,1-diphosphonicacid, tetrasodium salt(EHDP)and 2 rats remained untreated. Twelve hours prior to preparation on d 7, two of the EHDP-treated rats received an intraperitoneal injection of VD3 [1,25-(OH)_2 D_3] at 600 ng per kg body weight, while two rats did not receive the injection. Rats that did not receive VD_3 injection had decreased(P < 0.001) P absorption, but injection of VD3 resulted in increased(P < 0.001) absorption. The effect of VD3 injection was greater(P < 0.001) for rats fed 0.2% P diet than rats fed 0.8% P diet in ileum. Sodium dependent phosphate cotransporter type IIb(Na/Pi-IIb) and 25-hydroxyvitamin D 1-α hydroxylase(CYP27 B1) mRNA level showed the same trend with P absorption. Serum concentration of VD3 and la-hydroxylase activity in rats fed 0.2% P diet were lower than those fed 0.8% P diet. The injection of VD3 increased(P < 0.001)serum concentration of VD3 and la-hydroxylase activity. Thus, VD3 increased Na/Pi-Ⅱb and CYP27 B1 mRNA level and improved serum concentration of VD3 and la-hydroxylase activity in rats fed low-P diets.
