【摘要】： 目的探討白細(xì)胞介素10對(duì)腎小管上皮細(xì)胞轉(zhuǎn)分化的影響,并初步討論其作用機(jī)制。 方法本實(shí)驗(yàn)采用HK-2細(xì)胞為靶細(xì)胞。將HK-2細(xì)胞完全隨機(jī)分為5組:①對(duì)照組:5.5mmol/L的D-葡萄糖組;②高糖組:30mmol/L的D-葡萄糖組;③l ng/ml IL-l0組:30mmol/L的D-葡萄糖+ l ng/ml IL-l0組;④5 ng/ml IL-l0組:30mmol/L的D-葡萄糖+ 5 ng/ml IL-l0組;⑤25 ng/ml IL-l0組:30mmol/L的D-葡萄糖+ 25 ng/ml IL-l0組。培養(yǎng)48h后,倒置顯微鏡觀察HK-2細(xì)胞的形態(tài)變化;免疫細(xì)胞化學(xué)法檢測(cè)各組細(xì)胞E-cad、α-SMA蛋白的表達(dá);RT-PCR法檢測(cè)各組細(xì)胞PDGF mRNA、CTGF mRNA的表達(dá);western blot法檢測(cè)各組細(xì)胞PDGF、E-cad、α-SMA蛋白的表達(dá);ELISA法檢測(cè)各組細(xì)胞上清液中FN的含量。 結(jié)果(1)細(xì)胞形態(tài):對(duì)照組細(xì)胞呈鵝卵石樣緊密排列生長(zhǎng);高糖組部分細(xì)胞間隙增大,和附近的細(xì)胞脫離接觸,失去鵝卵石樣形態(tài),呈梭形;l ng/ml組、5 ng/ml組、25 ng/ml IL-l0組細(xì)胞形態(tài)介于對(duì)照組和高糖組之間。(2)高糖組與對(duì)照組相比,E-cad表達(dá)減少,PDGF、CTGF、α-SMA和FN表達(dá)增加,其差異均有顯著性(P0.05),提示高糖可誘導(dǎo)HK-2細(xì)胞轉(zhuǎn)分化。(3)l ng/ml組、5 ng/ml組、25 ng/ml IL-l0組與高糖組相比,E-cad表達(dá)增加,PDGF、CTGF、α-SMA和FN表達(dá)減少,其差異均有顯著性(P0.05),提示IL-10可以抑制HK-2細(xì)胞轉(zhuǎn)分化。 結(jié)論高糖可通過誘導(dǎo)PDGF和CTGF的表達(dá)使HK-2細(xì)胞E-cad表達(dá)減少、α-SMA和FN表達(dá)增加,從而誘導(dǎo)HK-2細(xì)胞轉(zhuǎn)分化及細(xì)胞外基質(zhì)產(chǎn)生;IL-10可以通過抑制PDGF和CTGF的表達(dá)從而抑制高糖誘導(dǎo)的HK-2細(xì)胞轉(zhuǎn)分化及細(xì)胞外基質(zhì)產(chǎn)生。
[Abstract]:Objective to investigate the effect of interleukin 10 (IL 10) on renal tubular epithelial cell transdifferentiation and its mechanism. Methods HK-2 cells were used as target cells. The HK-2 cells were randomly divided into 5 groups: 1 control group: 5.5 mmol / L D- glucose group; (2) high glucose group: 30 mmol / L D- glucose group; 30 mmol / L D- glucose group; 1 / 30 mmol / L D- glucose / L ng/ml IL-l0 group; 45 ng/ml IL-l0 group: 30 mmol / L D- glucose 5 ng/ml IL-l0 group; Ng/ml IL-l0 group: 30 mmol / L D- glucose 25 ng/ml IL-l0 group. After 48 hours of culture, the morphological changes of HK-2 cells were observed by inverted microscope. The expression of E-cadand 偽 -SMA protein was detected by immunocytochemistry and the expression of PDGF mRNA-CTGF mRNA was detected by RT-PCR. The expression of 偽 -SMA protein in the supernatant of each group was detected by Western blot, and the expression of 偽 -SMA protein was detected by Elisa. Results (1) Cell morphology: in the control group, the cells were closely arranged in pebbles, while in the high glucose group, the gap between the cells and the nearby cells was increased, and the pebble-like morphology was lost. The expression of E-cad, 偽 -SMA and FN in the high glucose group was significantly lower than that in the control group, and the expression of E-cad, 偽 -SMA and FN in the high glucose group was significantly lower than that in the control group, and the expression of 偽 -SMA and FN was increased in the high glucose group compared with the control group. The differences were significant (P0.05), suggesting that high glucose could induce the transdifferentiation of HK-2 cells. (3) the expression of E-cad increased in the 25 ng/ml IL-l0 group of 5 ng/ml group compared with the high glucose group, and the expression of E-cad, 偽 -SMA and FN decreased significantly (P0.05), suggesting that IL-10 could inhibit the transdifferentiation of HK-2 cells. Conclusion High glucose can reduce the expression of E-cad and increase the expression of 偽 -SMA and FN in HK-2 cells by inducing the expression of PDGF and CTGF. Thus, the transdifferentiation of HK-2 cells and the production of IL-10 in extracellular matrix can inhibit the transdifferentiation and extracellular matrix production of HK-2 cells induced by high glucose by inhibiting the expression of PDGF and CTGF.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類號(hào)】：R392.11

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 陳建糧,李曉玫,王海燕;白細(xì)胞介素10抑制核因子kB介導(dǎo)的腎小管上皮細(xì)胞表達(dá)細(xì)胞間黏附分子1[J];中華腎臟病雜志;2000年01期

2 李濤,冷希圣,秦致中,宋盛晗,趙力,熊亮發(fā),彭吉潤(rùn);白細(xì)胞介素10對(duì)肝星狀細(xì)胞激活的調(diào)節(jié)[J];中華肝臟病雜志;2005年01期

，

本文編號(hào)：2147951