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OBJECTIVE:To explore the anti-osteoporotic mechanism of icariin based on osteoblast and osteoclast proteomics. METHODS: The cell proliferation was determined by MTT assay. The activity of alkaline phosphatase (ALP) in osteoblasts and tartrate-resistant acid phosphatase (TRAP) in osteoclast were measured using ELISA kit. Alizarin red staining was used to observe the formation of bone mineralized nodules in osteoblasts. The number and area of bone resorption pit formed on bone slices by osteoclasts were used to characterize the activity of osteoclastic bone resorption. Two-dimensional gel electrophoresis was used to observe the changes of icariin-treated osteoblast and osteoclast proteomics. TOF/MS/MS analysis was applied to identify the differentially expressed proteins. RESULTS: Icariin significantly enhanced the osteoblastic bone formation and inhibited the osteoclastic bone resorption, and up-regulated or down-regulated expression of nine proteins in osteoblast and osteoclast, respectively. CONCLUSION: Icariin regulates bone metabolism through involving multiple biological process, including energy metabolism, oxidative stress, purine synthesis and cell growth and differentiation.
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Objective@#To compare the differentially expressed proteins in mice with kidney-yang deficiency and those with kidney-yin deficiency induced by hydrocortisone, and explore the similar and different material bases of male infertility caused by the two types of kidney deficiency.@*METHODS@#Thirty Kunming mice were equally randomized into a normal control, a kidney-yang deficiency and a kidney-yin deficiency group. The animals of the normal control group were injected intraperitoneally with normal saline at 0.2 ml qd for 7 days, while those of the latter two groups with hydrocortisone at 25 mg/kg/d for 10 days and 50 mg/kg/d for 7 days, respectively, for establishment of kidney-yang deficiency and kidney-yin deficiency models. Then the pathological changes in the testicular tissue of the mice were observed by HE staining and the differentially expressed proteins were compared among different groups using isobaric tags for relative and absolute quantitation (iTRAQ) and the bioinformatics method.@*RESULTS@#Sod1 was found to be a reproduction-related node protein differentially expressed in the testis tissues of the two types of kidney-deficiency mice, more highly expressed in the kidney-yin than in the kidney-yang deficiency group (P < 0.05). Five reproduction-associated node proteins were co-expressed in the testes of the two groups of kidney-deficiency mice, with significantly up-regulated expression of Rps28 and down-regulated expressions of Rpl11, Rplp2, Svs2 and Svs3a (P < 0.01).@*CONCLUSIONS@#Sod1 may be one of the key material bases for the differentiation of male infertility caused by kidney-yang deficiency from that induced by kidney-yin deficiency, while Rps28, Rpl11, Rplp2, Svs2 and Svs3a may be the common material bases of male infertility caused by the two types of kidney deficiency.
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Differential proteomics analysis of Sika deer antlers at rapid growth stage (60 d) and ossification stage ( 90 d) was performed by isobaric tags for relative and absolute quantitation ( iTRAQ ) , ultra high performance liquid chromatography and mass spectrometry technologies. A total of 127 differential proteins were identified. Compared with the ossification stage, 80 differential proteins were significantly up-regulated and 47 differential proteins were significantly down-regulated at the rapid growth stage. These differential proteins were mainly distributed in the regions of extracellular matrix, nucleosome, haptoglobin-hemoglobin complex, actin filament, endoplasmic reticulum-Golgi intermediate compartment, endoplasmic reticulum lumen, and endometrium, etc. The up-regulated differential proteins were mainly involved in the regulations of oxygen transport in the blood, nerve growth and regeneration, cartilage and bone development and ATP synthesis compared with ossification stage, and the down-regulated differential proteins were mainly involved in the endochondral ossification process. The changes of protein expression at different growth stages were closely related to antler rapid growth and ossification. Therefore, the results of this study provided a basic data for discovering the molecular mechanisms of antler rapid growth and ossification, and it was of great significance for further study of the pharmacological basis and clinical application of antlers.
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Objective To identify the specific proteins in two hydrophis (l#and 2#) with a simple differential proteins display method. Method By means of SDS-PAGE and Western blot, combined with analysis, the specific proteins in two hydrophis were obtained and been sequencing.Result The specific proteins of l#are 14.6kD and 8.7kD components and of 2#are 16.7kD. 10.2 kD components. The amino acid sequences of 14.6 kD and 10.2 kD components were assayed respectively. Conclusion Differential proteins can be obtained and used to identify the Chinese traditional medicinal crops by means of one-dimensional electrophoresis combined with antigen analysis .