| Objective:1.To observe the clinical efficacy of Gu Wei I Prescription for treating primary osteoporosis of kidney deficiency and blood stasis,and explore the significance of component.2.To observe the effects on rat bone microstructure of Gu Wei I Prescription through the animal experiment,and the expression of Wnt3a/?-catenin signaling pathways,to explore the mechanism in the treatment of osteoporosis of kidney and circulation invigorate drugs.Methods:Clinical study:50 patients with primary osteoporosis of kidney deficiency and blood stasis were selected from the outpatient and admission to the ward in the Second Affiliated Hospital of Nanjing University of Traditional Chinese Medicine(No.2 Traditional Chinese Medicine Hospital of Jiangsu Province),who were confirmed by DEXA and related clinical examination,and voluntarily joined the clinical study.They were divided into control group and treatment group,each has 25 cases.During their hospitalization,they were detected blood biochemical indicators(ALP,serum calcium and phosphorus)?bone metabolic markers(?-CTX?P1NP)and assessed the syndrome integral,and discharged after 1 week.After discharge,they continue to take oral Chinese medicine and anti-osteoporosis western medicine,reexamined VAS pain scores at 1 week,1 month and 3 months.At the time of 1 month after taking medicine,they were still need to review the blood biochemistry,syndrome integral,and effect evaluation.And at the time of 3 month after taking medicine,they were need to review the DEXA and bone metabolic markers.Experimental research:1.To observe the influence of Gu Wei ? Prescription on simulated osteoporosis rats,according to peripheral blood biochemistry,bone density,bone histology,bone mechanics and HE pathological morphology.2.To observe the influence of Gu Wei ? Prescription on simulated osteoporosis rats,according to BMP2?Runx2?Opn?Wnt3a mRNA in bone tissue by way of qPCR and Western Blotting experiment.Results:Clinical research section(1)47 cases(treatment group:24,control group:23)were eventually completed.The general data that are comparablehad no significant statistical difference between the treatment group and control group before treatment(P?0.10),and the back pain VAS score between two groups had no significant statistical differences(P?0.05).Blood serum ALP,serum calcium and phosphorus levels also had no significant statistical difference(P?0.05),and there was no significant difference between two group's kidney and blood stasis of TCM syndrome score,(P?0.05).(2)Changes in VAS scores after treatment:VAS scores in the treatment group and the control group did not decrease significantly at 1 week after treatment(all P>0.05),while VAS scores after 1 and 3 month decreased gradually and significantly,compared with those before treatment(P?0.05).There was no statistically significant difference between the two groups at 1 week after treatment(P?0.05),while there was statistically significant difference between VAS in the TCM treatment group at 1 and 3 month after treatment.(3)Changes in DXA after treatment:DXA were significantly higher than before treatment(P?0.05),and comparison between two groups show that there was no significant statistical difference between the two groups(P?0.05).(4)Changes in blood biochemistry(serum ALP,blood calcium,and blood phosphorus levels)after treatment:serum ALP,blood calcium,and blood phosphorus levels were significantly higher than before treatment(P?0.05),and comparison between two groups show that the serum ALP,blood calcium and blood phosphorus levels in the treatment group were significantly higher than those in the control group after treatment,with statistically significant differences(P?0.05).(5)Changes in bone metabolic markers(?-CTX?P1NP)after treatment:Bone metabolic marker(?-CTX)was significantly lower than before treatment(P?0.01),P1NP was significantly higher than before treatment(P?0.01),and comparison between two groups show that the bone metabolic markers(?-CTX?P1NP)in the treatment group were have more significantly changer than those in the control group after treatment,with statistically significant differences(P?0.01).(6)Patients in two groups had better satisfaction with the efficacy after treatment,with 91.67%in the treatment group and 82.61%in the control group.The comparison between the two groups showed that the treatment group was significantly better than the blank group.(7)After 1 month of treatment,both groups could improve the patients' TCM syndrome score of kidney deficiency and blood stasis(P?0.05).In comparison,the improvement of the score in the treatment group was significantly better than that in the control group,and there was a significant statistical difference between the two groups(P?0.05).Experimental research section:Compared with the blank group,the bone density?maximum stress and maximum load of rats in the model group after oophorectomy for 5 weeks was significantly reduced,and their bone volume fraction BV/TV,trabecular number Tb.N,trabecular thickness Tb.Th were also significantly reduced,but trabecular separation degree Tb sp was significantly increased.Pathological examination of HE also showed that the bone lesions of the rats were obvious,the tissue structure was diseased and the morphology was incomplete.Model group rats' serum calcium,serum phosphorus concentrations,creatinine were decreased,and serum alkaline phosphatase(ALP),blood urea nitrogen(BUN),alanine aminotransferase(ALT)and aspartate aminotransferase(AST)rise.These difference were statistically significant,who indicated that bone metabolism was in a state of high transition and osteoporosis model was replicated successfully.(2)Gu Wei I Prescription has significant curative effect on osteoporosis,each dose group could significantly increased bone mineral density level,maximum stress and maximum load,increased the BV/TV,Tb.N,Tb.Th.The serum calcium,phosphorus and creatinine increased,while ALP,BUN,ALT and AST decreased,compared with model group.These index present dose dependence.The pathological examination of HE also found that the bone morphology was improved after treatment.(3)The protein levels of BMP2,Runx2,Opn and Wnt3a/?-catenin signaling pathway in bone tissues of osteoporosis model rats were all lower than those in the normal blank group,suggesting that abnormal Wnt3a/?-catenin signaling pathway in bone tissues was closely related to the incidence of osteoporosis.(4)qPCR and WB tests both found BMP2,Runx2,Opn and Wnt3a/?-catenin signaling pathway protein levels rise,prompt that Gu Wei I Prescription could promot Wnt3a/?-catenin pathway transcription through mediation,thus promote the differentiation of osteoblast proliferation,in order to achieve the corresponding therapeutic effect.Conclusion:Gu Wei I Prescription can effectively relieve the patients' pain of thoracic?lumbar and back,who suffered primary osteoporosis with kidney and blood stasis type.The prescription combined with conventional anti-osteoporosis drug treatment could increase serum ALP,serum calcium,serum phosphorus levels,and change?-CTX?P1NP,which are important indicators of bone metabolism.After treatment,the patients had better satisfaction with the curative effect in general,and could effectively improve the TCM syndrome score of kidney deficiency and blood stasis,and compared with single western drug treatment,the effect is more significant.Therefore,this prescription have good clinical efficacy,and it reflect that TCM dialectical treatment of kidney deficiency and blood stasis osteoporosis has its advantages.Through animal experiments,we also found that Gu Wei I Prescription can promoted Wnt3a protein and BMP2,Runx2,Opn factor through regulating Wnt3a/?-catenin signaling pathway,to promote the differentiation of osteoblast proliferation and achieve the curative effect of osteoporosis. |
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