| Objectives1. Clinical research partIn this part, the objective is to observe the effects on glucose metabolism and bone metabolism on diabetic osteoporosis by using ZiShen JiangTang Pill to treat these patients. And to provide a modern and scientific theoretical basis for ZiShen JiangTang Pill treating diabetic osteoporosis.2. Experimental research partIn this part, in order to investigate the preventive and therapeutic effect of ZiShen JiangTang Pill on bone mass loss and formation of osteoporosis which were caused by diabetes and the possible molecular mechanism, we use the pill to cure diabetic rats to observe the influence on bone metabolism index, bone mineral density and form of bone tissue of these rats, and to study the regulatory effect of ZiShen JiangTang Pill for Wnt/β-catenin signal pathway.Methods1. Clinical research part40 cases of diabetic osteoporosis with Kidney Yin Deficiency Syndrome were selected between January,2015 and April,2015 in Shenzhen Hospital of Traditional Chinese Medicine, and randomly divided into the control group of 20 cases, and the treatment group of 20 cases. The control group was additionally given alendronate sodium, based on the basic glucose-lowering treatment, while the treatment group was additionally given ZiShen JiangTang Pill based on the treatment given to the control group. The course of treatment was three months. TCM syndrome integral, bone mineral density, fasting plasma glucose (FPG), fasting insulin (Fins), serum calcium, serum phosphorus, osteocalcin (OC) and β-cross-linked c-terminal telopeptide of type1 collagen (β-CTX) were detected.2. Experimental research partThe rat model of diabetes was established by intraperitoneal injection of streptozotocin (STZ). The rates were randomly divided into model group, ZiShen JiangTang Pill treated group, strontium ranelate group. And the corresponding therapy was given through gastric tube for rats in different treatment group. Meanwhile, a control group was set in normals animals. There were 10 rats in each group. After 3 months, bone mineral density, FBG, Fins, serum calcium, serum phosphorus, bone alkaline phosphatase (B-ALP),OC, total procollagen type 1 N-propeptide (PINP),β-CTX and osteoprotegerin (OPG) were detected. At the same time, the changes of form of bone tissue also been observed by HE staining, and the gene expression level of Wnt3, low density lipoprotein peceptor related protein-5 (LRP-5), β-catenin, runt-related transcripttion factor-2 (RUNX-2), dickkopf-1 (DKK-1) and sclerostin (SOST) were tested by quantitation real-time PCR (QRT-PCR)Results1. Clinical research part(1) Bone mineral density, Fins, serum calcium, serum phosphorus and OC of the treatment group were all remarkably increased after treatment (P＜0.01), meanwhile, the total mark of TCM syndrome, FPG and β-CTX were remarkably decreased (P＜0.01);(2) Bone mineral density, Fins, serum phosphorus and OC of the control group were all significantly increased after treatment (P＜0.05 or P＜0.01), at the same time, FPG and β-CTX were significantly decreased (P＜0.01), however, the total mark of TCM syndrome and serum calcium had no obvious changes (P ＞0.05);(3) The levers of the above indexs had significantly difference between groups after treatment and the comparison of difference before and after treatment of the above indexs between groups also had statistical significance (P＜ 0.05 or P＜0.01), furthmore, the improvement degree of the above indexs of the treatment group were superior to the control group.2. Experimental research part(1) Compared with the normal control group, Bone mineral density, Fins, serum calcium, serum phosphorus, B-ALP and OC were all significantly decreased (P ＜0.01), while FPG, PINP, β-CTX and OPG were significantly increased in the rats of the model group(P＜0.01); In contrast to the model group, Bone mineral density, Fins, serum calcium, serum phosphorus, B-ALP and OC were all significantly increased (P＜0.01), while FPG, PINP, β-CTX and OPG were significantly decreased in the rats of the ZiShen JiangTang Pill treated group and, the strontium ranelate group (P＜0.01); Compared with the strontium ranelate group, Fins, serum phosphorus, OC, PINP and OPG were all significantly increased (P＜0.05, or P＜0.01), while FPG and β-CTX were significantly decreased in the rats of the ZiShen JiangTang Pill treated group (P＜0.01), however, bone mineral density and serum calcium had no obvious changes (P ＞0.05);(2) The arrangement of bone trabecula of rats in the normal control group was tight and ordered, and the texture was clear, meanwhile, the thickness of bone cortexhad was normal, and both of bone trabecula and cortex had no obvious pathological changes; while the structure of bone trabecula of rats in the model group became sparse, some even fracture, the number of bone trabecula decreased, and bone cortex became thin; meanwhile compared with the model group, the structure disturbance of bone tissue of rats in the ZiShen JiangTang Pill treated group and the strontium ranelate group had been obviously improved and was nearly normal;(3) Compared with normal control group, the gene expressive quantity of Wnt3, LRP-5, β-catenin and RUNX-2 of bone tissue of rats in the model group were obviously decreased (P＜0.01); while compared with the model group, the gene expressive quantity of Wnt3, LRP-5, β-catenin and RUNX-2 in the ZiShen JiangTang Pill treated group were significancantly increased (P＜0.01), at the same time, the gene expressive quantity of β-catenin and RUNX-2 in the strontium ranelate group were also significancantly increased (P＜0.01), however the gene expressive quantity of Wnt3 and LRP-5 had no obvious changes (P＞0.05); Compared with the strontium ranelate group, the gene expressive quantity of Wnt3 and LRP-5 were significancantly increased (P＜0.01), however the gene expressive quantity of β-catenin and RUNX-2 had no obvious changes (P＞0.05); And there were no significant differences of the gene expressive quantity of DKK-1 or SOST among the four groups (P＜0.05)Conelusions1. Clinical research partZiShen JiangTang Pill not only can improve clinical symptoms and physical signs of diabetic osteoporosis, but also can improve the glucose metabolism disorder, reduce the loss of serum calcium and phosphorus, then improve bone metabolism, promote bone formation, inhibit bone resorption, and increase bone mineral density.2. Experimental research part(1) There is conduction disturbance in Wnt/β-catenin signal pathway in bone tissue of diabetes rats because of down-regulation of the gene expressive quantity of Wnt3, LRP-5, β-catenin and RUNX-2, thus effecting the formation and differentiation of osteoblast, reducing the formation of bone, destructing the form of bone tissue, and decrease bone mineral density eventually;(2) Zishen JiangTang Pill can up-regulate the gene expressive quantity of the above important molecular, then promote the formation and differentiation of osteoblast, inhibit bone destruction, improve the form of bone tissue, increase bone mineral density, regulate bone metabolism disorder and play a role in preventive and therapeutic effect on bone mass loss and the formation of osteoporosis of diabetes rats. |
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