内毒素对NRK52E细胞生长及细胞间缝隙连接功能的影响

【目的】 研究不同浓度内毒素(LPS)对NRK52E细胞生长和细胞间缝隙连接功能的影响&#65377;【方法】 体外培养NRK52E细胞,随机分为对照组及LPS处理组,LPS处理组分别用5个不同浓度的LPS(10 ng/mL&#65380;50 ng/mL&#65380;100 ng/mL&#65380;500 ng/mL&#65380;1 000 ng/mL)作用24 h&#65377;采用四甲基偶氮唑盐法检测NRK52E细胞的生长;细胞荧光免疫示踪法测定NRK52E细胞间缝隙连接传递的功能;Western Blotting测定对照组&#65380;LPS 10 ng/mL组和LPS 100 ng/mL组Cx43蛋白的表达&#65377; 【结果】 ① 与对照组和LPS 10 ng/mL组相比,LPS 50 ng/mL&#65380;100 ng/mL&#65380;500 ng/mL及1 000 ng/mL组细胞生长明显降低(P < 0.01)&#65377;②与对照组相比,LPS 100 ng/mL&#65380;500 ng/mL及1 000 ng/mL组细胞间缝隙连接功能明显降低(P < 0.01)&#65377;③内毒素浓度与NRK52E细胞生长和细胞间缝隙连接传递数目呈负相关(r = -0.941,-0.872,P < 0.01)&#65377;④与对照组比较,LPS 10 ng/mL组和LPS 100 ng/mL组的Cx43蛋白表达明显降低(P < 0.05)&#65377; 【结论】 LPS对NRK52E细胞的生长呈浓度相关性的抑制,这种作用与细胞间缝隙连接功能有关&#65377;

Effects of Endotoxin on Growth Activity and Gap Junction of NRK52E Cells

Objective]This study was designed to observe the effects of endotoxin(LPS)on the growth activity and gap junction(GJ)of NRK52E cells.Methods]The NRK52E cells were divided into control group and LPS groups,and the NRK52E cells in LPS groups were treated with LPS 10 ng/mL,50 ng/mL,100 ng/mL,500 ng/mL,and 1 000 ng/mL for 24 h respectively.The NRK52E cells growth activity was measured through MTT method,and the function of gap junction of NRK52E cells was measured through the method of fluoroimmunoassay.The protein expression of connexin 43(Cx43)in control group,LPS 10 ng/mL group,and LPS 100 ng/mL group were also determined by Western blotting.Results]①Compared with control group and LPS 10 ng/mL group,The NRK52E cells growth activity decreased significantly in LPS 50 ng/mL,100 ng/mL,500 ng/mL,and 1 000 ng/mL groups(P＜0.01).②Compared with control group,the function of GJ decreased significantly in LPS 100 ng/mL,500 ng/mL,and 1 000 ng/mL groups(P＜0.01).③There were negative correlations among the concentration of LPS and NRK52E cells growth activity and GJ function respectively(r=-0.941,-0.872,P＜0.01).④Compared with control group,the protein expression of Cx43 were decreased significantly in LPS 10 ng/mL and 100 ng/mL groups(P＜0.01).Conclusions]LPS can inhibit the NRK52E cells growth activity in a dose-depend manner.GJ function is one of the mechanisms.