Impact Of Bushenyijing Method On Type â…¡ Collagen And Proteoglycan Gene Expression During The Differentiation Of MSCs Into Cartilage Cells

Objective:Through the induction of mesenchymal stem cells (Mesenchymal Stem Cells, MSCs) to chondrogenic differentiation, the representative observed Bushenyijing side Zuoguiwan containing serum on proliferation during the differentiation of MSCs, the expression of chondrocyte marker genes typeâ…¡collagen and proteoglycan Expression, in order that "the main bone of kidney,"molecular mechanisms and from traditional Chinese medicine to find bone cartilage protective agent basis. Methods:1.Use whole bone marrow method (adherence screening method) separation and culture of rat MSCs and identify;2.Preparation Zuoguiwan containing serum, Using HPLC method to preliminary analysis the composition of Zuogui Pill containing serum, to get stable interference drugs;3.CCK-8 assay Zuoguiwan containing serum on the proliferation of MSCs;4. MSCs induced to differentiate into chondrocytes, RT-PCR, immunohistochemistry and Real-time PCR analysis of differentiation of chondrocytes induced typeâ…¡collagen and proteoglycan expression.Results:1.Separation of whole bone marrow adherent MSCs, flow cytometry showed high expression of CD13 and CD105 in the third generation,but not the expression of CD33,CD34, CD38,CD45, consistent with stem cell characteristics,21d can get around more purified MSCs;2. HPLC analysis of serum, to detect stable Zuoguiwan's Jun drug Rehmannia glutinosa the main component of 5-hydroxymethylfurfural (5-hydroxymethyl-2-furfura,5-HMF) components, suggesting the intervention of reliable drugs;3.Different doses of Zuoguiwan containing serum could promote proliferation of MSCs, in which the proliferation of the most obvious medium dose group(p<0.05);4. Induced 21 days RT-PCR and immunohistochemistry showed the expression of MSCs in typeâ…¡collagen mRNA and protein, which Zuoguiwan containing serum groupâ…¡collagen mRNA expression was significantly higher than that induced group; Real-time PCR showed that in the 21d Zuoguiwan containing serum group, the relative expression of proteoglycan mRNA quantification is 7d and 14 were expressed about the amount of 16 times and 3 times (p<0.05), and induction was significantly higher than the control group (p<0.05).Conclusion:Promote the proliferation of MSCs andâ…¡collagen,proteoglycan gene expression may be one of cartilage protective effect of kidney medicine molecular basis Keywords:Zuogui Pill; Mesenchymal Stem Cells;Cartilage...