Neuroscience letters · 2023
Effects of RNA m6A writer METTL3 and hDPSCs on the peripheral nerve regeneration: In vitro and in vivo study.
СтатьяПоддержка исследования: вне правительства СШАНазвание и аннотация: Перевод в очереди
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Hei Wei-Hong, Du Ming-Yuan, He Hong
Аннотация
PURPOSE: This study aimed to investigate whether RNA m6A participated in the differentiation and proliferation of dental pulp stem cells and improved peripheral nerve regeneration using a rat model of crushed mental nerve injury.
MATERIALS AND METHODS: The components of RNA m6A were analyzed through qRT-PCR, while cell proliferation of different groups, including over-expression METTL3 (OE-METTL3) hDPSCs group, knock-down METTL3 (KD-METTL3) hDPSCs group and hDPSCs group in vitro, was clarified by MTT assay. Five groups were designed, namely, Control group, Sham group, hDPSCs group, OE-METTL3 group and KD-METTL3 group. After crushed right mental nerve injury, cells of different groups were transplanted into the lesion area (6 ul in volume). At one, two and three weeks later, histomorphometric analysis and sensory test were conducted in vivo.
RESULTS: qRT-PCR results showed that "METTL3" was participated in the differentiation of dental pulp stem cells. There were differences (P < 0.05) between OE-METTL3 group and control group in MTT results in the third, fourth and sixth days. Moreover, the sensory test revealed significant differences (P < 0.05) in difference score and gap score between OE-METTL3 group and KD-METTL3 group in the first and third weeks. The axon counts and retrograde labeled neurons significantly increased in OE-METTL3 group compared with KD-METTL3 group.
CONCLUSIONS: These results demonstrated that RNA m6A participated in the differentiation and proliferation of dental pulp stem cells, and that OE-METTL3 group exhibited the greater ability to improve peripheral nerve regeneration than KD-METTL3 group and hDPSCs group.
MATERIALS AND METHODS: The components of RNA m6A were analyzed through qRT-PCR, while cell proliferation of different groups, including over-expression METTL3 (OE-METTL3) hDPSCs group, knock-down METTL3 (KD-METTL3) hDPSCs group and hDPSCs group in vitro, was clarified by MTT assay. Five groups were designed, namely, Control group, Sham group, hDPSCs group, OE-METTL3 group and KD-METTL3 group. After crushed right mental nerve injury, cells of different groups were transplanted into the lesion area (6 ul in volume). At one, two and three weeks later, histomorphometric analysis and sensory test were conducted in vivo.
RESULTS: qRT-PCR results showed that "METTL3" was participated in the differentiation of dental pulp stem cells. There were differences (P < 0.05) between OE-METTL3 group and control group in MTT results in the third, fourth and sixth days. Moreover, the sensory test revealed significant differences (P < 0.05) in difference score and gap score between OE-METTL3 group and KD-METTL3 group in the first and third weeks. The axon counts and retrograde labeled neurons significantly increased in OE-METTL3 group compared with KD-METTL3 group.
CONCLUSIONS: These results demonstrated that RNA m6A participated in the differentiation and proliferation of dental pulp stem cells, and that OE-METTL3 group exhibited the greater ability to improve peripheral nerve regeneration than KD-METTL3 group and hDPSCs group.