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The application discloses a method for semi-quantitative analysis of expression level of secreted protein of human joint cartilage cells by using immunoblotting technology. The method comprises the following steps: 1) preparing a secreted protein culture solution; 2) collecting a sample of the secreted protein culture solution; 3) performing dialysis desalting treatment on the sample; 4) concentrating the secreted protein culture solution to prepare a powder; 5) re-dissolving the secreted protein culture solution powder and quantifying the protein; 6) separating the protein components in the re-dissolved culture solution; and 7) detecting the expression level of the secreted protein in the re-dissolved culture solution sample by using immunoblotting method. The low-temperature dialysis and vacuum concentration technology can significantly improve the concentration of the secreted protein in the human joint cartilage tissue culture solution sample under the condition of maintaining the integrity of the protein structure, and is suitable for detecting the chemical fluorescence signal of the secreted protein in the sample by using the immunoblotting technology. The application provides an important cell reaction detection tool for studying the pathogenesis of bone and joint diseases caused by degenerative changes of cartilage tissue and researching and developing biological small-molecule drugs for treating the diseases.