Mechanism of DAIa2GIP inhibiting mitochondrial apoptosis in chondrocytes

doi:10.3969/j.issn.2095-4344.3093

Abstract

Abstract: BACKGROUND: The previous research of our group has shown that DAIa2GIP, a new analogue of glucose dependent insulin stimulating polypeptide, has a protective effect on chondrocytes, but its mechanism is not clear.
OBJECTIVE: To observe the effect of DAIa2GIP on interleukin-1β (IL-1β) induced apoptosis of chondrocytes, and to explore its molecular biological mechanism.
METHODS: The costal chondrocytes of Sprague-Dawley rats were extracted and identified by SABC immunohistochemistry. The third generation cells were divided into six groups: (1) normal control group; (2) IL-1β induction group; (3) DAIa2GIP+IL-1β induction group; (4) DAIa2GIP+IL-1β+pro3GIP (GIPR antagonists) induction group; (5) DAIa2GIP induction group; (6) pro3GIP induction group. After 48 hours of drug treatment, mitochondrial membrane potential was tested, cell apoptosis was detected using Annexin-V FITC Kit (phosphatidylserine eversion analysis), calcium overload of cells was determined under laser confocal microscope, and cytochrome C content were measured by ELISA. The study protocol was approved by the Animal Ethics Committee of Shanxi Medical University.
RESULTS AND CONCLUSION: SABC immunohistochemical staining showed that collagen II could be developed as chondrocytes; the maintenance degree of mitochondrial membrane potential was significantly higher in the IL-1β+DAIa2GIP incubation group than the IL-1β induction group, but the potential in both groups was lower than that in the normal control group. The apoptotic rate of chondrocytes in the IL-1β+DAIa2GIP incubation group was significantly lower than that in the IL-1β induction group, but the apoptotic rate in both groups was higher than that in the normal control group. The degree of calcium overload was compared with fluorescence intensity, and the result showed that the IL-1β+DAIa2GIP incubation group had a significantly higher intensity than the IL-1β induction group, and the fluorescence intensity in both groups was higher than that in the normal control group. The release of chondrocyte mitochondrial cytochrome C in the IL-1β+DAIa2GIP incubation group was significantly lower than that in the IL-1β induction group (P < 0.01), and the release amount in both groups was significantly higher than that in the normal control group (P < 0.01). The above four indicators showed no significant difference between the IL-1β induction group and the DAIa2GIP+IL-1β+pro3GIP group, but the values in both groups were lower than those in the control group. There was no significant difference among the normal control group, DAIa2GIP induction group, and pro3GIP induction group. To conclude, DALa2GIP can effectively antagonize IL-1β induced mitochondrial dysfunction of rat chondrocytes, thus antagonizing chondrocyte apoptosis. In this process, DALa2GIP can effectively reduce the degree of calcium overload and the release of cytochrome C.

Key words: bone">, cartilage">, osteoarthritis">, chondrocyte">, mitochondria">, apoptosis">, calcium overload">, cytochrome C

CLC Number:

Wang Yue, Wang Xinjun, Yuan Yinpeng, Wang Yuze. Mechanism of DAIa2GIP inhibiting mitochondrial apoptosis in chondrocytes[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(11): 1652-1657.

Figures/Tables 5

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