| Objcetive1.A clinical randomized controlled trial of positive drug control was used to objectively evaluate the clinical efficacy of a comprehensive regimen of Bu Shen Qiang Du Zhi Lv Decoction in improving Renal Deficiency Deficiency Ankylosing Spondylitis bone metabolism,revealing its bone protection potential and providing a large sample size for the later period.The center's randomized controlled trials provide a reference.2.Based on the network pharmacology to predict the therapeutic pathways and targets of bone metabolism regulation of Ankylosing spondylitis(AS)in the treatment of ankylosing spondylitis(AS),which lays a foundation for subsequent research.3.Based on the research results of network pharmacology,a preliminary experiment was conducted to verify the molecular mechanism of Bu Shen Qiang Du Zhi Lv Decoction prescription in regulating AS bone metabolism.Methods1.Clinical studyA clinical randomized controlled trial was used to include 34 patients with ankylosing spondylitis of the Kidney Deficiency Deficiency Syndrome of TCM Syndrome.The patients were divided into TCM and Western medicine control groups,and functional exercise was used as the basic treatment.The experimental group was treated with Bu Shen Qiang Du Zhi Lv Decoction and subtraction,and the control group was treated with Celecoxib capsules.Through 24-week clinical observation,?-CTX,N-MID,P1 NP,ASDASCRP,CRP,ESR,BASDAI,BASFI,BASMI were used.DXA,MRI and other indicators to evaluate the Bu Shen Qiang Du Zhi Lv comprehensive Program to improve the clinical efficacy of ankylosing spondylitis bone metabolism and joint function,imaging,explore its bone protection potential.2.Network pharmacologyWith the aid of network pharmacology,we preliminary explored and analyzed the possible correlation between the potential target protein of Bu Shen Qiang Du Zhi Lv Decoction and ankylosing spondylitis.It was found that the biological effects of Bu Shen Qiang Du Zhi Lv Decoction Compound on the treatment of AS are mainly related to the regulation of inflammation and related pathological changes.The network of target proteins interacting with AS-BSQD is mainly tumor necrosis factor signaling pathway,estrogen signaling pathway,and ovarian steroidogenesis.Signaling pathways,T cell receptor signaling pathways,osteoclast differentiation pathways,rheumatoid arthritis signaling pathways,cytokine-cytokine receptor interaction signaling pathways,NF-kappa B signaling pathways,and Screening the AS-BSQD intersection target protein HUB GENE,we obtained 23 HUB GENE and found that the HUB GENE pathways are mainly concentrated on immune and inflammatory signaling pathways,hormone signaling pathways,cancer signaling pathways,and cytokine signaling pathways.Direct pathways associated with bone metabolism include osteoclast differentiation pathways,Wnt signaling pathways,rheumatoid arthritis signaling pathways,and NF-kappa B signaling pathways.3.Experimental StudyUsing in vitro experimental methods,SD rat osteoblasts were cultured in vitro.MTS method was used to observe the effect of Bushen Qiang Du Zhi Lv Decoction on the proliferation of osteoblasts;alkaline phosphatase activity assay was used to detect the kidney strengthening supervision.Effects of Sputum Decoction on Alkaline Phosphatase Secretion of Osteoblasts in Rats;Western Blotting Method for Exploring Bu Shen Qiang Du Zhilv Decoction for Osteoblast TNF-?,c-Fos,OPG,RANKL,TRAF2,DKK1 Effect of protein expression.Effect of Bu Shen Qiangling Recipe on Anti-apoptosis Expression of Osteoblast Inflammation Model Induced by TNF-?.Results1.Clinical studyThe Bu Shen Qiang Du Zhi Lv comprehensive Program can reduce the main efficacy index ?-CTX,decrease serum N-MID,ALP in the bone metabolic index,CRP in inflammation index,BASMI in disease function index,improve thorax activity,and imaging index Hip BMD,Lumbar BMD,Left femoral neck BR,Left femoral intertrochanteric BR,Left femoral stem BR,Ankle edema(SPARCC score),PGA score in patient's own disease assessment,ASQo L in quality of life assessment Scores,SDS scores,self-control before and after treatment,the difference was not statistically significant(P>0.05);P1NP in serum bone metabolic markers could be elevated,and there was no significant difference between the two groups before and after treatment(P>0.05);There was no improvement in BASFI in functional indicators(P>0.05).The Bu Shen Qiang Du Zhi Lv comprehensive Program can improve ASDASCRP in disease activity of ankylosing spondylitis,BASDAI in disease function index,ESR in inflammation index,VAS score in patient's own disease evaluation,TCM symptom score in quality of life assessment,SF-36 indicators,before and after treatment their own control,the difference was statistically significant(P<0.05).Western medicine can reduce the main efficacy indicators of ?-CTX,serum bone metabolism indicators of N-MID,P1 NP,ALP,disease function indicators of BASFI,BASMI,improve thorax activity,imaging parameters of lumbar BMD,left femur There was no significant difference in cervical SR,SF-36,SDS,TCM symptom scores in quality of life evaluation before and after treatment(P>0.05),and no improvement in inflammation index CRP and imaging parameters SPARCC scores.(P>0.05).Western medicine comprehensive program can improve disease activity ASDASCRP,BASDAI in disease function index,ESR in inflammation index,PGA,VAS score in patient's own disease evaluation,left femoral intertrochanteric BR in imaging index,left femur stem BR,ASQo L score in quality of life evaluation,selfcontrol before and after treatment,the difference was statistically significant(P<0.05).All indicators,the experimental group and the control group had no significant difference between the two groups(P>0.05).2.Network pharmacologyWith the aid of network pharmacology,we preliminary explored and analyzed the possible correlation between the potential target protein of Bu Shen Qiang Du Zhi Lv Decoction and ankylosing spondylitis.It was found that the biological effects of Bu Shen Qiang Du Zhi Lv Decoction Compound on the treatment of AS are mainly related to the regulation of inflammation and related pathological changes.The network of target proteins interacting with AS-BSQD is mainly tumor necrosis factor signaling pathway,estrogen signaling pathway,and ovarian steroidogenesis.Signaling pathways,T cell receptor signaling pathways,osteoclast differentiation pathways,rheumatoid arthritis signaling pathways,cytokine-cytokine receptor interaction signaling pathways,NF-kappa B signaling pathways,and the like.Screening the AS-BSQD intersection target protein key genes,we obtained 23 HUB GENE and found that the HUB GENE pathways are mainly concentrated on immune and inflammatory signaling pathways,hormone signaling pathways,cancer signaling pathways,and cytokine signaling pathways.Direct pathways associated with bone metabolism include osteoclast differentiation pathways,Wnt signaling pathways,rheumatoid arthritis signaling pathways,and NF-kappa B signaling pathways.3.Experimental StudyThe Bu Shen Qiang Du Zhi Lv Decoction Compound has a promoting effect on the proliferation of rat osteoblasts.At 72 hours,the proliferation rate of osteoblasts in the low-concentration group(12.5ng/ml)can reach 1.85±0.18(P<0.01).Bu Shen Qiang Du Zhi Lv Decoction can promote the secretion of alkaline phosphatase in rat osteoblasts,and it can reach the highest peak on the 9th day of dosing culture,and increase on the 14 th day compared with the positive control group.The difference was statistically significant(P<0.01~0.001).The ratio of OPG/RANKL in the low(12.5,25,50 ng/ml)and middle dose(100 ng/ml)groups of Bu Shen Qiang Du Zhi Lv Decoction Prescription was higher than that of the positive control group,with a statistically significant difference(P<0.05~ 0.001).Compared with the celecoxib-positive control group,the expression of TRAF2 protein was significantly decreased in the low-,middle-,and high-dose groups of Bu Shen Qiang Du Zhi Lv Decoction(P<0.05~0.001).Compared with the positive control group,the expression of c-Fos protein was significantly decreased in the Bu Shen Qiang Du Zhi Lv Decoctionlow-,middle-,and high-dose groups(all P<0.001).Compared with the positive control group,the DKK1 protein expression was upregulated in the high-dose group(800ng/ml)of Bu Shen Qiang Du Zhi Lv Decoction(P<0.01),and DKK-was lower in the low-dose group(50ng/ml)than in the positive control group.1 The protein expression decreased(P<0.05);Bu Shen Qiang Du Zhi Lv Decoction can reduce the apoptosis rate of osteoblast inflammation model,in which the low dose group(50ng/ml)apoptosis rate decreased compared with the positive control group,the difference was statistically significant Significance(P<0.01).Conclusion1.The Bu Shen Qiang Du Zhi Lv comprehensive Program can improve the bone metabolism of ankylosing spondylitis of kidney deficiency type,improve inflammation,improve disease function index,ankle edema,quality of life of patients,and TCM symptom scores,and has certain bone protection potential.Better security.2.The results of the network pharmacological study suggest that: Bu Shen Qiang Du Zhi Lv Decoction Compound Formula may act on AS through signaling pathways such as inflammation,hormones,osteoclast metabolism,and immune factors.Direct pathways associated with bone metabolism include osteoclast differentiation pathways,Wnt signaling pathways,rheumatoid arthritis signaling pathways,and NF-kappa B signaling pathways.3.Different doses of Bu Shen Qiang Du Zhi Lv Decoction compound compound acting on AS through the osteoclast pathway and Wnt signaling pathway cannot not only inhibit the bone destruction of AS,but also inhibit osteoblasts on the molecular level at the same time.Over-differentiation to avoid local heterotopic ossification. |
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