Membrane Ion Channel Blockers And The CSK Undermine The Influence Of Tensile Stress To Stimulate The In Vitro Metabolism Of Physiological Functions Of Rabbit Articular Chondrocytes

Objective:Under tensile stress conditions, CSK disrupting agents and membrane ion channel blockers on the metabolism of chondrocytes.Methods:2months in New Zealand white rabbit only10, germ-free separation under the condition of double knee joint cartilage, and randomly divided into8groups, the control group, microtubule, microfilament damage group, middle fiber damage group, Na+, K+, Ca2+and Cl-channel block group. In tensile stress,(tensile rate was10%, the frequency of0.5Hz, sine wave,6h/D) under the action of load, three days later, respectively, to join the cytoskeleton breaker breaker, breaker of actin filaments, microtubules and the middle fiber breaker and ion channel blockers (Na+, K+, Ca2+and Cl-channel blockers),37℃to3D. Using HE staining to observe their morphology; EL AS A to detect each group in the cell culture supernatant on MMP-3, TIMP-1protein content; Alcian blue staining method to detect the contents of each medium, GAG; RT-PCR method to detect each group of cells in the Col-Ⅱ, MMP-3, TIMP-1, Act, Tub, Vim relative expression of mRNA.Results:Compared with control group, the cytoskeleton damage group and cartilage cells form ion channels block group were arranged unevenly, forms and nuclei are deep, size is differ, the phenomenon such as cytoplasm reduced; And GAG concentration in the supernatant on cytoskeleton damage group than the control group decreased, with statistical significance (P<0.05); Protein levels in tests showed that microfilament, concentration of MMP-13middle fiber damage group than the control group decreased, with statistical significance (P<0.05), the middle fiber damage concentration was significantly increased in group of TIMP-1, was statistically significant (P<0.05); Ca2+, Cl-ion channels block group GAG concentration than the control group decreased, with statistical significance; K+, Cl-channel block MMP-13 concentration than the control group were lower, there is statistical significance (P<0.05); Genetic level test results consistent with the testing results of protein levels; Each experimental group chondrocytes Col-Ⅱ relative expression of mRNA were cut, there is statistical significance (P<0.05).Conclusion:Under the effect of tensile stress, the damage CSK and blocks the ion channels are making cartilage degradation, including microtubules, the middle fibers and Na+and Cl-channel may be closely related with metabolism of cartilage matrix synthesis.