Research On Theory And Related Experiments Of Growth And Development Of Kidney Main Bone Based On Cx43 Protein

ObjectiveTo clarify the theoretical connotation of traditional Chinese medicine of "kidney main bone growth and development" and the situation of modern medical research.The Cx43 protein and its mediated GJIC function are used as the entry point for theoretical discussion.Experimental studies were conducted on rat BMSCs cultured cartilage to induce differentiation in vitro to demonstrate the regulatory effect and mechanism of Zuogui--wan on Cx43 protein.Methods1 、 Theoretical research part: Theoretical research clarifies the motherland medicine’s understanding of the theory of "kidney main bone growth and development" from the perspective of theoretical origin,theoretical connotation,and theoretical application.According to the research on "kidney main bone growth and development" in modern medicine,the essence of modern medicine in theory is discussed.2、Experimental study part:SPF grade 2-month-old SD male rats with traditional Chinese medicine concentrate wereintragastrically administered,and a serum-containing serum culture medium with a volume fraction of 10% was prepared by serum pharmacological methods for co-cultivation.Use density gradient centrifugation to prepare BMSCs for culture and passage,add inducing solution to induce chondrogenesis.Experimental grouping: set up a normal control group A(medium added with the inducing solution prepared by the administration of animal saline infused with saline),and a group B of Zuoguiwan medicated serum(induced solution prepared by the medium with Zuoguiwan medicated serum).Pill-containing serum+ gap junction block group C(same as group B + AGA).After grouping the cells,the medium was changed every 3 days and induced to culture for 21 days.Cell morphology was observed during the cultivation process;on the 14 th day,RT-PCR technology was used to detect the expression of Cx43 and GDF-5mRNA,immunofluorescence staining was used to detect the expression of Cx43 protein,and the stain dye tracer method was used to detect the cell GJIC function;on the 21 st day,the immune group Chemical detection of type II collagen expression.Detection of various indicators,comparison between groups.The data obtained are all expressed in SPSS 17.0 software.One-way analysis of variance was used for comparison between groups,using t test.P<0.O5 means the difference is statistically significant.Result1、Theoretical research part: The theory of kidney main bone growth and development is derived from the "kidney main bone" theory in the "The Yellow Emperors Internal Classic"."Kidney stores essence,produces bone marrow and nourishes bone" is thetheoretical core of it."Qi-blood-essence-marrow " is closely related,they are important material components that form the external shape of bone.“Shen yin” and “Shen yang”are based on the essence of kidney and coordinate with each other to maintain normal bone growth and development.Kidney metaplasia provides the material basis for bone growth and development;under pathological conditions,kidney deficiency affects bone growth and development,bone motor function,and produces various bone diseases.Modern medical research has proved that there is a close relationship between kidney and bone in traditional Chinese medicine.The process of "kidney main bone growth and development" is specifically reflected in the regulation of the dynamic balance between the NEI network,various bone cells and the microenvironment.Among them,BMSCs research,Cx43 protein and GJIC function-mediated research are important research contents in the substantive research of "kidney main bone growth and development".2、Experimental study part: The cells of the cartilage to differentiation induction culture change from the initial fusiform shape to a circle or a round shape.The cytoplasm is rich and secretes a large amount of matrix to differentiate into chondrocytes.The serum containing Zuoguiwan can significantly promote the expression of Cx43 mRNA and GDF-5mRNA.Compared with group A and group B,P<0.O5.Cx43 protein fluorescent reaction deposits are mainly expressed on the surface of the cell membrane,cytoplasm,and near the nucleus.The average integrated optical density value is compared between groups.Zuoguiwan-containing serum can significantly promote theexpression of Cx43 protein in cells.Group B is compared with group A,P<0.O5.In the GJIC dye labeling test,it can be seen that green fluorescence diffuses on both sides of the scratch,and there is a difference between the groups in the quantitative analysis of fluorescence.Compared with groups A and C,P<0.O5.Cell type II collagen is expressed in the cytoplasm and extracellular matrix.The average optical density of the positive reactants is statistically significant between the groups.The difference between group B and group A is P<0.O5.After adding AGA,the expression of Cx43 mRNA and protein and GDF-5mRNA had no significant effect,but the expression of type II collagen decreased.Quantitative analysis showed that group B was significantly higher than groups A and C,P<0.O5.ConclusionModern medicine has proved in many ways that there is a close relationship between kidney and bone in traditional Chinese medicine.Cx43 protein may be an important material basis for "kidney main bone growth and development" and an important functional level of kidney essence.Zuogui wan-containing serum can increase the expression of Cx43 protein and GJIC function during the differentiation of BMSCs into cartilage,and the developmental gene GDF-5 plays a synergistic role in this process.This may be an important mechanism in promoting the differentiation of BMSCs into cartilage.