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本研究探讨对非肌性肌球蛋白IIA的表达和功能,以阐明MYH9综合征肾脏病变和中性粒细胞包涵体形成的机制。采用半定量Western-blot检测中性粒细胞非肌性肌球蛋白IIA的表达;以胚肾细胞(HEK-293)为研究对象对IIA、IIB之间的相互作用进行了探讨;RT-PCR法检测HEK-293细胞中非肌性肌球蛋白IIA、IIB的表达,并对其进行了免疫共沉淀的研究。结果显示:先证者中性粒细胞IIA/β-actin比值为(0.35±0.12),较正常对照中性粒细胞IIA/β-actin的比值(0.87±0.18)明显减少(p<0.01)。非肌性肌球蛋白IIA、非肌性肌球蛋白IIB在HEK-293细胞中均有较高的表达;免疫共沉淀分析表明,非肌性肌球蛋白IIA和非肌性肌球蛋白IIB均有表达,而阴性对照两者均无表达。结论:非肌性肌球蛋白IIA的负显性效应导致了中性粒细胞包涵体的产生。IIA和IIB有明显的相互作用,IIB至少是部分补偿了Fechtner综合征突变的IIA蛋白的作用并延缓了组织的功能障碍。
This study explored the expression and function of non-myosin IIA to elucidate the mechanisms of nephropathy and neutrophil inclusion formation in MYH9 syndrome. Semi-quantitative Western-blot was used to detect the expression of neutrophil non-myosin IIA; The interaction between IIA and IIB was investigated by embryonic kidney (HEK-293) cells; RT-PCR The expression of non-myosin IIA and IIB in HEK-293 cells was detected and co-immunoprecipitation was performed. The results showed that the ratio of IIA / β-actin in neutrophils was (0.35 ± 0.12), and the ratio of IIA / β-actin in neutrophils was significantly decreased (p <0.01). Non-myosin IIA, non-myosin IIB in HEK-293 cells were higher expression; co-immunoprecipitation analysis showed that non-myosin IIA and non-myosin IIB are There was no expression of the negative control. Conclusion: The negative dominant effect of non-myosin IIA results in the production of neutrophil inclusion bodies. There is a clear interaction between IIA and IIB, with IIB at least partially compensating for the role of IIA protein in the mutation of Fechtner’s syndrome and delaying tissue dysfunction.