白细胞介素6反义RNA对肾小球系膜细胞的影响

Regulation of recombinant interleukin-6 antisense RNA on glomerular mesangial cell

为了观察白细胞介素６（ＩＬ－６）反义ＲＮＡ对肾小球系膜细胞（ＭＣ）的影响。方法将ＩＬ０６反义ＲＮＡ用脂质体导入包装细胞ＰＡ３１７，经Ｃ４１８筛选，ＰＣＲ技术分析，并用＾３Ｈ－胸腺嘧啶（＾３Ｈ－ＴｄＲ）掺入法，ＫＤ６３细胞株，Ｎｏｒｔｈｅｒｎ杂交技术分别检测ＩＬ－６反义ＲＮＡ对肾小球细胞增殖、分泌及ＩＬ－６以及表达ＩＬ－６ｍＲＮＡ的影响。结果重组假病毒滴度为１×１０＾７ＣＦＵ／ｍｌ。

Objective To determine whether the recorabinant murine interleukin-6 antisense RNA (IL-6 RNAAS) can regulate the proliferation of murine glomerular mesangial cell (MC) and the expression of protein and IL-6 mRNA that derived from the MC. Methods IL-6 RNAAS was led into the package cell PA31; by lipofect AMINE. Presence of IL-6 RNAAS in the supernatant of the cultural PA3n cells was assured by G4u screening and PCR check. The MC proliferation, IL-6 activity and IL-6 mRNA expressionin of MC were detected by 3 H-thymidine, KD3,7 cell line and Northern blot respectively. Results The pse.udovirus liter in the PA317 cell supernatant reached 1 ×107 CFU/ml. The IL-6 RNAAS could incorporate into MC and was expressed. The 3H-thymidine incorporation, IL-6 mRNA expression and MC-derived IL-6 were significantly lower than that in control group. Conclusion IL-6 RNAAScan inhibit MC proliferation and regulate downward the secretion and expression of IL-6 in MC.