The Basic Studies Of Tonifying The Kidney On GIOP And Analysis Of Correlation With The Syndrome Types

ObjectiveThis study is to explore the mechanism of preventing the GIOP by Tonifying the Kidney. In this Retrospective clinical study, the analysis of existing patient medical records of syndromes features is implemented, distributed regularity of Chinese syndromes of GIOP is foud out, at the same time, combined with risk factors research to further explore the advantages of TCM prevention and treatment of GIOP with Tonifying the Kidney. In our animal experiments, Bone metabolism index, bone mineral density(BMD),Micro-CT index, Bone biomechanical parameters and Bone LRP5、RUNX2、RANKL、NF-κB、β-catenin mRNA were assayed to explain the mechanism and effects of preventing the GIOP by Tonifying the Kidney in rat. The combination of Chinese and western prevention GIOP was to provide the basis of clinical and experimental efficacy.Methods1、118 cases are randomly selected by The first affiliated hospital of Guangzhou university of CM, Spine surgery special inpatient department visits in the period from January 2014 to January 2015. A retrospective survey, record the patient’s general condition, including gender, ethnicity, age, height, weight, body mass index (BMI), as well as the performance of symptoms and the physical and chemical examination, including dual-energy X-ray absorptiometry acquisition lumbar bone mineral density, which in turn from the general risk factors between TCM distributing for analyzes. Statistical analysis data use SPSS 17.0 software package for statistical analysis.2、 Sprague Dawley rats (N= 45) were divided randomly into five groups: Blank control group(BC),Model group(M),Western medicine group(WM),Chinese medicine group(CM), Chinese and Western medicine group(WM-TCM).After establishing the GIOP model in all rats. The rats were oral fed with Distilled Water in Untreated Control group, the rats were subcutaneous injection with Prednisone Dragon5 mg/kg/d, the rats were oral fed with Allen Sodium Phosphate of lmg/kg/7d in Western medicine group, the rats were oral fed with Tonifying the Kidney Prescription of 2.5 g/kg/d in Chinese medicine group, the rats were oral fed with Allen Sodium Phosphate and Tonifying the Kidney Prescription respectively. Anesthesia is executed after 12 weeks of rats.2.1 Detection of bone metabolic indices:Detecting the bone formation markers (PINP), and bone resorption markers(β-CTx) expression in serum of rats by ELISA.2.2 Detection of BMD:After rats death, quickly completely stripping lumbar 1-3 vertebral body from the body, and testing lumbar 1-3 overall bone mineral density (BMD), bone mineral content (BMC),with dual-energy X-ray absorptiometry bone mineral density (hologic companies in the United States).2.3 Detection of Micro-CT:The lumbar vertebral cancellous bone microstructure parameters were obtained by scanning the lumbar 4 vertebrae of the rats with The μ CT80 type micro-CT scanning machine.2.4 Detection of Detection of bone biomechanical parameters:The compression test was carried out on lumbar 4 vertrbrae of rats with the universal material testing machine (Electropuls E1000 test system, Instron Corporation, USA), determinating the compression stiffness (N/mm) and the compression strength (N) of rats vertebrae2.5 Detection of LRP5、RUNX2、RANKL、NF-κB、β-catenin mRNA:Detecting the LRP5、RUNX2、RANKL、NF-KB、β-catenin mRNA in lumbar 6 vertrbrae and left tibia of rats with QT-PCR tests. Reference to TAKARA company extraction kit, reverse transcription kit and fluorescent quantitative kit.Results1、This study found that The CM type of GIOP were grouped Kidney Yang deficiency(27.12%), Kidney Ying deficiency(28.81%), Kidney Ying and Yang deficiency(10.17%), Qi and Xue stasis (19.49%), Spleen and Stomach deficiency(14.41%) types, Kidney deficiency type was the main type, and the characters of Kidney deficiency type displayed obviously as the time follew. The fracture rate of kidney deficiency patient reaches up to 80.64%, which higher than other two groups.2、Animal experimental results are as follows:2.1 Detection of bone metabolic indices:Compared to the model group, BC group, WM group, TCM group and WM-TCM group have significant difference (P<0.05) at the expression of β-CTX and PINP, while there is no significant difference between among WM group, TCM group and WM-TCM group (P>0.05).2.2 Detection of BMD and BMC:Compared to the model group, BC group, WM group, TCM group and WM-TCM group have significant difference (P<0.05) at BMC and BMD. But the WM group、TCM group and WM-TCM group has no significant difference compared to BC group (P>0.05).2.3 Detection of Micro-CT:Compared to the model group, BC group, WM group, TCM group and WM-TCM group have significant difference (P<0.05) at Tb. Sp and SMI, along with significant difference between BC group and TCM group (P<0.05).2.4 Detection of bone biomechanical parameters:Compressive compressive strength of BC group, WM group, TCM group and WM-TCM group have significant difference compared to the model group (P<0.05); meantime compared to the model group it has statistical differences in WM group and WM-TCM (P<0.05).2.5 Detection of LRP5、RUNX2、RANKL、NF-KB、β-catenin mRNA:Compared to the model group, the other four groups have significant difference (P<0.05) at the expression of LRP5% RUNX2、RANKL, while which have no significant difference at NF-KB、β-catenin mRNA (P>0.05).Conclusion1、This study found that The CM type of GIOP were grouped Kidney Yang deficiency, Kidney Ying deficiency, Kidney Ying and Yang deficiency, Qi and Xue stasis and Spleen and Stomach deficiency types, Kidney deficiency type was the main type, and the characters of Kidney deficiency type displayed obviously as the time follew. The proportion of patients with kidney deficiency type fracture was 80.64%, higher than other two groups the incidence of syndrome types. The method of tonifying the kidney plays an important pole in preventing and treating GIOP and its complications. Therefore, clinical prevention and treatment should be strengthened with the combination of Chinese and western medicine on GIOP and its complications.2、According to the animal experiment results, some conclusions can be advised following:2.1 The Tonifying Kidney Prescription can raise the expression level of PINP and reduce the expression level of β-CTX in the GIOP rats serum, to promote osteogenesis, inhibit the action of the osteoclasts.2.2 The Tonifying Kidney Prescription can significantly increase bone mineral density in the GIOP rats; which had exact curative effect on the prevention of rat GIOP; In addition, the application of The Tonifying Kidney Prescription combined with Allen sodium phosphate on bone mineral density of the GIOP rats was better than using Allen sodium phosphate or kidney party alone.2.3 The Tonifying Kidney Prescription could increase the GIOP rats trabecular number, separating degree and reduce bone structure model index, so as to improve bone quality and bone strength, which could prevent the happening of the GIOP.2.4 The Tonifying Kidney Prescription could obviously improve the rat bone compression stiffness (N/mm) and the compression strength (N), then improving the GIOP rats biomechanical properties so as to prevent the GIOP in rats.2.5 The Tonifying Kidney Prescription can improve the gene expression of LRP5 and RUNX2mRNA, restrain the gene expression of RANKLmRNA in GIOP rats, and reflect its promote osteogenesis, prevention of GIOP in rats.