BACKGROUND To study the role of lysosomal decomposition and elimination of old bone matrix, as well as the mechanism of promoting chondrocyte growth and bone recovery through the perspective of TFEB-mediated lysosomal autophagy. METHODS Rat models of acute knee injury were designed, and autophagy flow was detected by injection of autophagy inhibitors 3-methyladenine. Autophagy flow was detected by RFP-GFP-LC3 double fluorescence molecule. The expression of TFEB, DRAM, MAPLC3, and MITF were analyzed by Western blot, and the expression of genes NITF, Bcl2, and TYR in rat cartilage tissues were detected by RT-PCR. RESULTS The number of autophagosomes was increasing in the auto group compared with the inhibitor-auto group and normal group. There was a significant difference of LC3 levels in the auto group and inhibitor-auto group compared with the normal control. The expression of TFEB, DRAM, MAPLC3, and MITF proteins by Western blot analysis were significantly increased in the auto group and decreased in the inhibitor-auto group. The expression of NITF, Bcl2, and TYR by RT-PCR determination were higher in the auto group and inhibitor-auto group than the normal group. CONCLUSIONS Autophagy can inhibit apoptosis, promote chondrocyte growth and bone regeneration, and restore knee joint injury of rats. The main mechanism is to promote the effect of TFEB-mediated lysosomal autophagy.

中文翻译：

---

溶酶体自噬通过TFEB介导促进急性膝关节损伤大鼠的恢复。

背景技术从TFEB介导的溶酶体自噬的角度研究溶酶体分解和消除旧骨基质的作用，以及促进软骨细胞生长和骨骼恢复的机制。方法设计大鼠急性膝关节损伤模型，注射自噬抑制剂3-甲基腺嘌呤检测自噬流量。通过RFP-GFP-LC3双荧光分子检测自噬流。Western blot分析TFEB，DRAM，MAPLC3和MITF的表达，RT-PCR检测大鼠软骨组织中NITF，Bcl2和TYR的表达。结果与抑制剂-自动组和正常组相比，自动组中自噬体的数量正在增加。自动组和抑制剂-自动组的LC3水平与正常对照组相比有显着差异。通过蛋白质印迹分析，TFEB，DRAM，MAPLC3和MITF蛋白的表达在自动组中显着增加，而在抑制剂-自动组中降低。逆转录-聚合酶链反应（RT-PCR）测定的NITF，Bcl2和TYR的表达在自动组和抑制剂自动组中高于正常组。结论自噬可以抑制大鼠的细胞凋亡，促进软骨细胞的生长和骨骼再生，并能恢复大鼠膝关节的损伤。主要机制是促进TFEB介导的溶酶体自噬作用。通过RT-PCR测定的NITF，Bcl2和TYR的表达在自动组和抑制剂-自动组中高于正常组。结论自噬可以抑制大鼠的细胞凋亡，促进软骨细胞的生长和骨骼再生，并能恢复大鼠膝关节的损伤。主要机制是促进TFEB介导的溶酶体自噬作用。通过RT-PCR测定的NITF，Bcl2和TYR的表达在自动组和抑制剂-自动组中高于正常组。结论自噬可以抑制大鼠的细胞凋亡，促进软骨细胞的生长和骨骼再生，并能恢复大鼠膝关节的损伤。主要机制是促进TFEB介导的溶酶体自噬作用。