Study on connection between tensile force-activated ion channel and F-actin of cytoskeleton

doi:10.3877/cma.j.issn.1674-134X.2020.06.008

Abstract

Abstract:

Objective

To study the connection between the new mechanically-activated(MA) cation channel protein (Piezo1) and the cytoskeleton of the human osteoarthritis (OA) chondrocytes under mechanical tensile force by using a Flexercell unit.

Methods

The primary human chondrocytes were isolated, cultured, and then subjected to the mechanical tensile force for zero, two, 12, 24 and 48 h, respectively. The expressions of Piezo1 in each group were assessed by reverse transcription-polymerase chain reaction(RT-qPCR). The immunofluorescence was used to prove that the Piezo1 could be expressed in the human chondrocytes, and the location of the Piezo1 protein. The Piezo1 inhibitor GsMTx4 was used to block the cation channel of Piezo1, served as a positive control. The relative expression levels of the RT-qPCR were compared by one-way ANOVA, and the comparison between the two groups was performed by q test.

Results

The results of the RT-qPCR showed that the relative mRNA expression of Piezo1 in the two-hour group was higher than that in the blank group without statistical significance (P>0.05). The 12 h group was significantly higher than the blank group (F=9.785, P<0.05). The expression level of the 24 h group was the highest, and that of the 48 h group was decreased. After GsMTx4 treatment, the expression of the Piezo1 significantly decreased in all the groups (F=13.907, P<0.001). The results of laser scanning confocal microscopy showed that the cytoskeleton of chondrocytes gradually thickened and concentrated from the two-hour group to the 24 h group, but the cytoskeleton of chondrocytes in 48 h group was loose and degraded. The corresponding RT-qPCR results showed that the expression of Piezo1 mRNA had the same trend with the concentration and looseness of cytoskeleton. The expression of GsMTx4 decreased in all the groups.

Conclusion

The expression of the new mechanical cation channel Piezo1 protein shows that it is time-dependence under the tensile force in the human chondrocytes and the activation of the Piezo1 protein could be triggered by the change of the cytoskeleton of the human chondrocytes.

Key words: Ion channels, Chondrocytes, Actins, Stress, mechanical

Xiaofei Li, Yi Sun, Zhao Zhang, Haining Zhang. Study on connection between tensile force-activated ion channel and F-actin of cytoskeleton[J]. Chinese Journal of Joint Surgery(Electronic Edition), 2020, 14(06): 691-697.

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Figures/Tables 5

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