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目的:观察高糖对肾小管上皮细胞凋亡和内质网应激标志物表达的影响,研究冬虫夏草提取液对高糖诱导的肾小管上皮细胞内质网应激的可能保护机制。方法:体外培养大鼠肾小管上皮细胞,分为正常低糖组、低糖+冬虫夏草组、高渗组、高渗+冬虫夏草组、高糖组、高糖+冬虫夏草组和高糖+高渗+冬虫夏草组,用Realtime-PCR法检测各组细胞葡萄糖调解蛋白78(Grp78)、半胱氨酸蛋白酶蛋白12(caspase-12)及I型胶原mRNA的表达,采用酶联免疫吸附法测定细胞上清液中I型胶原的分泌,Western-blotting检测Grp78及caspase-12蛋白表达。结果:高糖组与低糖组相比,Grp78、Caspase-12及Ⅰ型胶原mRNA和蛋白水平明显上升,差异均有统计学意义(P<0.05);低糖+冬虫夏草组、高渗组及高渗+冬虫夏草组与低糖组相比,Grp78、Caspase-12 mRNA及Ⅰ型胶原mRNA和蛋白水平无明显差别(P>0.05);与高糖组相比,高糖+冬虫夏草干预组Grp78、Caspase-12及Ⅰ型胶原mRNA和蛋白表达下降,差异均有统计学意义(P<0.05)。结论:冬虫夏草对高糖诱导下肾小管上皮细胞凋亡及肾脏纤维化具有一定抑制作用,其机制可能与抑制内质网应激反应有关。
OBJECTIVE: To observe the effect of high glucose on the apoptosis of renal tubular epithelial cells and the expression of endoplasmic reticulum stress markers, and to explore the possible protective mechanism of Cordyceps sinensis extract on endoplasmic reticulum stress induced by high glucose in renal tubular epithelial cells. Methods: The renal tubular epithelial cells were cultured in vitro and divided into normal low glucose group, low glucose + Cordyceps sinensis group, hyperosmotic group, hypertonic + Cordyceps sinensis group, high glucose group, high glucose + Cordyceps sinensis group and high glucose + Real-time PCR was used to detect the expression of Grp78, caspase-12 and type I collagen mRNA in each group. The cell supernatants were determined by enzyme-linked immunosorbent assay Type I collagen secretion, Western-blotting detection of Grp78 and caspase-12 protein expression. Results: The mRNA and protein levels of Grp78, Caspase-12 and type Ⅰ collagen in high glucose group were significantly higher than those in low glucose group (P <0.05). Low glucose + Cordyceps sinensis group, hyperosmotic group and hypertonic group + Cordyceps sinensis group compared with low glucose group, Grp78, Caspase-12 mRNA and collagen type Ⅰ mRNA and protein levels no significant difference (P> 0.05); compared with high glucose group, high glucose + Cordyceps intervention group Grp78, Caspase-12 And type Ⅰ collagen mRNA and protein expression decreased, the differences were statistically significant (P <0.05). CONCLUSION: Cordyceps sinensis can inhibit renal tubular epithelial cell apoptosis and renal fibrosis induced by high glucose, which may be related to the inhibition of endoplasmic reticulum stress.