摘要
The effects of AAV-TGFβ1 and AAV-TGFβ3 on promoting synthesis of glycosaminoglycan and collagen type II of dedifferentiated rabbit lumbar disc NP cells were studied in this work. The rabbit lumbar disc NP cells were isolated and cultured. The earlier and later dedifferentiated NP cells were established by subculture. The AAV transfection efficiency to dedifferentiated NP cells was analyzed with AAV-EGFP in vitro. After dedifferentiated NP cells were transfected by AAV-TGFβ1 or AAV-TGFβ3, their biological effects on promoting synthesis of glycosaminoglycan or collagen type II were detected and compared by the methods of 35S incorporation or immunoblotting. The experimental results showed that AAV could transfect efficiently the earlier dedifferentiated NP cells, but its transfection rate was shown to be at a low level to the later dedifferentiated NP cells. Both AAV-TGFβ1 and AAV-TGFβ3 could promote the earlier dedifferentiated NP cells to synthesize glycosaminoglycan and collagen type II, and the effect of AAV-TGFβ1 was better than that of AAV-TGFβ3. For the later dedifferentiated NP cells, the AAV-TGFβ3 could promote their synthesis, but AAV-TGFβ1 could slightly inhibit their synthesis. Therefore, AAV-TGFβ1 and AAV-TGFβ3 could be used for the earlier dedifferentiated NP cells, and the TGFβ3 could be used as the objective gene for the later dedifferentiated NP cells.
The effects of AAV-TGF beta_1 and AAV-TGF beta_3 on promoting synthesis ofglycosaminoglycan and collagen type II of dedifferentiated rabbit lumbar disc NP cells were studiedin this work.The rabbit lumbar disc NP cells were isolated and cultured.The earlier and laterdedifferentiated NP cells were established by subculture.The AAV transfection efficiency todedifferentiated NP cells was analyzed with AAV-EGFP in vitro.After dedifferentiated NP cells weretransfected by AAV-TGFp,or AAV-TGF beta_3,their biological effects on promoting synthesis ofglycosaminoglycan or collagen type II were detected and compared by the methods of 35S incorporationor immunoblotting.The experimental results showed that AAV could transfect efficiently the earlierdedifferentiated NP cells,but its transfection rate was shown to be at a low level to the laterdedifferentiated NP cells.Both AAV-TGF beta_1,and AAV-TGF beta_3 could promote the earlierdedifferentiated NP cells to synthesize glycosaminoglycan and collagen type II,and the effect ofAAV-TGFp,was better than that of AAV-TGF beta_3.For the later dedifferentiated NP cells,the AAV-TGFbeta_3 could promote their synthesis,but AAV-TGFp,could slightly inhibit theirsynthesis.Therefore,AAV-TGFp,and AAV-TGF beta_3 could be used for the earlier dedifferentiated NPcells,and the TGF beta_3 could be used as the objective gene for the later dedifferentiated NPcells.
基金
Supported by the National Natural Science Foundation of China (Grant No. 30271318)
