Abstract
Objective. To investigate the effect of isoflurane (ISO) on the proliferation, apoptosis, and inflammatory response of lipopolysaccharide- (LPS-) induced normal human astrocytes (NHAs) by regulating the miR-206/BDNF axis. Methods. NHA proliferation activity was measured by MTT; NHA apoptotic rates were measured by Annexin V-FITC/PI; western blotting was used to measure the BDNF expression; ELISA was used to measure the IL-6, IL-1β, and TNF-α expression in NHAs; qPCR was used to measure the expressions of miRNAs that are related to NHAs proliferation and apoptosis; dual-luciferase reporter was constructed to validate the targeting relationship between miR-206 and BDNF. Results. LPS increased the proliferation activity and decreased the apoptosis rate of NHAs which were effectively reversed by the ISO (p<0.05); LPS significantly inhibited the expression of miRNAs related to proliferation and apoptosis in NHAs (p<0.05, p<0.01), whereas ISO significantly increased the expression of miR-206 (p<0.01) by downregulating the expression of BDNF, thus inhibiting NHA proliferation and inflammatory response and enhancing apoptosis. Conclusion. ISO can inhibit the expression of BDNF by upregulating the expression of miR-206, thereby inhibiting the proliferation and inflammatory response of NHAs and promoting its apoptosis.
Highlights
The incidence of neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS) and spinal cord injury (SCI) is increasing, and the central nervous system inflammatory response is a potential mechanism for various neurodegenerative diseases [1]
normal human astrocytes (NHAs) are involved in the development of a variety of neurodegenerative diseases [4]; studying the mechanism of reactive NHAs is helpful for the development of protective strategies for neurodegenerative diseases induced by central nervous system inflammation
MTT results showed that the cell proliferation ability increased in a dosedependent manner after treatment of NHAs with different concentrations of LPS (0.0 μg/mL, 0.2 μg/mL, 0.5 μg/mL, and 1.0 μg/mL) (Figure 1(a), p < 0:01), and there is no significant difference in cell proliferation activity when LPS was at a concentration of 0.5 μg/mL and 1 μg/mL
Summary
To investigate the effect of isoflurane (ISO) on the proliferation, apoptosis, and inflammatory response of lipopolysaccharide- (LPS-) induced normal human astrocytes (NHAs) by regulating the miR-206/BDNF axis. LPS increased the proliferation activity and decreased the apoptosis rate of NHAs which were effectively reversed by the ISO (p < 0:05); LPS significantly inhibited the expression of miRNAs related to proliferation and apoptosis in NHAs (p < 0:05, p < 0:01), whereas ISO significantly increased the expression of miR-206 (p < 0:01) by downregulating the expression of BDNF, inhibiting NHA proliferation and inflammatory response and enhancing apoptosis. ISO can inhibit the expression of BDNF by upregulating the expression of miR-206, thereby inhibiting the proliferation and inflammatory response of NHAs and promoting its apoptosis
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