Volume 16 Issue 1
Jul.  2022
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GU Yun-tao, CHEN Ke-wei, BIAN Yang-yang, FU Jian, YUAN Wei, XING Kong-ming, PENG Lei. Diazoxide prevents rat chondrocyte apoptosis via Inhibition of endoplasmic reticulum stress[J]. Chinese Journal of General Practice, 2018, 16(1): 14-18,37. doi: 10.16766/j.cnki.issn.1674-4152.000004
Citation: GU Yun-tao, CHEN Ke-wei, BIAN Yang-yang, FU Jian, YUAN Wei, XING Kong-ming, PENG Lei. Diazoxide prevents rat chondrocyte apoptosis via Inhibition of endoplasmic reticulum stress[J]. Chinese Journal of General Practice, 2018, 16(1): 14-18,37. doi: 10.16766/j.cnki.issn.1674-4152.000004

Diazoxide prevents rat chondrocyte apoptosis via Inhibition of endoplasmic reticulum stress

doi: 10.16766/j.cnki.issn.1674-4152.000004
  • Received Date: 2017-05-02
  • Objective To research the mechanism of diazoxide on hydrogen peroxide induced chondrocyte apoptosis. Methods We obtain the carilage cells from the 3 SD rat articular cartilages. The cartilage cells were divided into six groups, namely the control group(A), the H2O2-injured group(B), H2O2 + 100 μmol/L DZ group(C), H2O2 + 200 μmol/L DZ group(D), H2O2 + 300 μmol/L DZ group(E), H2O2 + 400 μmol/L(F). Group A had no special treatment, group B incubated with 400 μmol/L H2O2 for 8 hours in 37℃ incubator, the C, D, E group were pre-incubated with 100, 200, 300, 400 μmol/L DZ in a 37℃ incubator incubated 30 minutes, then incubated with 400 μmol/L H2O2 solution in 37℃ incubator. The activity of chondrocytes in group A, B, C, D, E and F were detected by CCK8 method. The apoptosis of chondrocytes were detected by flow cytometry. The function of chondrocytes were detected by RT-PCR and immunofluorescence, Western blot were used to detect the expression of endoplasmic reticulum stress proteins. Results ①Using the CCK8 assay, the order of the rate of cells activity:group A > group E > group D > group F > group B. ②The apoptosis rates of cells were detected by Flow cytometer. The order of apoptosis rates of chondrocyte as follow:group B > group F > group D > group E > group A. ③The Coll-Ⅱ protein expression of chondrocyte were detected by RTPCR analysis, as follow:group A > group E > group D > group F > group B. However, the aggrecanase expression Contrary to Coll-Ⅱ. ④the expression of CHOP in each chondrocytes group detected by immunofluorescence, as follow:group B > group F > group A. ⑤The Caspase-3, Bax, CHOP protein expression of chondrocyte were detected by Westernblot analysis, as follow:group B > group F > group A. Conclusion Diazoxide prevents H2O2-induced rat chondrocyte apoptosis via inhibition of endoplasmic reticulum stress.

     

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