【摘要】：目的:通過抑制磷酸化絲/蘇氨酸激酶(phosphorylation serine/threonine kinase,p-Akt)的表達,觀察并探討p-Akt和端粒酶逆轉(zhuǎn)錄酶(telomerase reverse transcriptase,TERT)對高糖高脂干預(yù)的胰島β細胞凋亡的影響。方法:將NIT-1細胞隨機分成對照組(正常培養(yǎng))、Akt抑制組(正常培養(yǎng)+Akt抑制劑SH5)、高糖高脂干預(yù)組(高糖高脂培養(yǎng))、高糖高脂+Akt抑制組(高糖高脂培養(yǎng)+Akt抑制劑SH5),TUNEL法和流式細胞術(shù)檢測各組細胞凋亡,Western blot檢測各組細胞TERT、Akt和p-Akt蛋白表達水平,ELISA檢測各組細胞胰島素分泌。結(jié)果:與正常對照組比較[凋亡率:(2.88±1.25)%;胰島素分泌:(0.65±0.10);p-Akt:(0.80±0.07);TERT:(1.63±0.12)];高糖高脂干預(yù)組胰島β細胞凋亡率(38.25±8.63)%明顯增加,胰島素分泌(0.40±0.09)下降,p-Akt(0.55±0.05)和TERT(1.03±0.05)表達明顯下降,差異均有統(tǒng)計學(xué)意義(P0.05);與高糖高脂干預(yù)組[凋亡率:(38.25±8.63)%;胰島素分泌:(0.40±0.09);p-Akt:(0.55±0.05);TERT:(1.03±0.05)]比較,高糖高脂+Akt抑制組細胞凋亡率(62.00±7.13)%明顯增高,胰島素分泌(0.23±0.10)下降,p-Akt(0.28±0.07)和TERT(0.61±0.07)表達明顯下降,差異均有統(tǒng)計學(xué)意義(P0.05)。結(jié)論:抑制p-Akt表達可增加高糖高脂誘導(dǎo)的胰島β細胞凋亡,其機制可能與降低了TERT表達水平有關(guān)。
[Abstract]:Aim: to observe and investigate the effects of p-Akt and telomerase reverse transcriptase (telomerase reverse transcriptase,TERT (telomerase reverse transcriptase,TERT) on apoptosis of islet 尾 cells treated with high glucose and high fat by inhibiting the expression of phosphorylated serine / threonine kinase (phosphorylation serine/threonine kinase,p-Akt). Methods: NIT-1 cells were randomly divided into control group (normal culture), Akt inhibitor SH5), high glucose and high fat intervention group (high glucose and high fat culture), high glucose and high fat Akt inhibition group (high glucose and high fat Akt inhibitor SH5). TUNEL assay and flow cytometry were used to detect the expression of TERT,Akt and p-Akt protein, and ELISA was used to detect the insulin secretion in the cells of each group. The expression of TERT,Akt and p-Akt protein was detected by, Western blot. Results: compared with the normal control group, the apoptosis rate was (2.88 鹵1.25)%, the insulin secretion rate was (0.65 鹵0.10), the expression of insulin was (0.80 鹵0.07); TERT: (1.63 鹵0.12), and the rate of insulin secretion was (0.65 鹵0.10). The apoptosis rate (38.25 鹵8.63)%, insulin secretion (0.40 鹵0.09) and the expression of p-Akt (0.55 鹵0.05) and TERT (1.03 鹵0.05) were significantly decreased in high glucose and high fat intervention group. The difference was statistically significant (P0.05); [apoptosis rate: (38.25 鹵8.63)%; insulin secretion: (0.40 鹵0.09); pu Akt: (0.55 鹵0.05)] and high glucose and high fat intervention group [apoptosis rate: (38.25 鹵8.63)%; insulin secretion: (0.40 鹵0.09); Compared with the control group, the apoptosis rate (62.00 鹵7.13)% and insulin secretion (0.23 鹵0.10) were significantly increased and insulin secretion decreased (0.23 鹵0.10) in the group of high glucose and high fat Akt inhibition (TERT:). The expression of p-Akt (0.28 鹵0.07) and TERT (0.61 鹵0.07) decreased significantly (P0.05). Conclusion: the inhibition of p-Akt expression can increase the apoptosis of islet 尾 cells induced by high glucose and high lipid, which may be related to the decrease of TERT expression level.
【作者單位】： 重慶市涪陵中心醫(yī)院體檢部;第三軍醫(yī)大學(xué)第一附屬醫(yī)院內(nèi)分泌科;
【基金】：國家自然科學(xué)基金資助項目(編號:81370885) 重慶市教委研究生科研創(chuàng)新資助項目(編號:CYB14097) 第三軍醫(yī)大學(xué)青年人才基金資助項目(編號:SWH2013QN02)
【分類號】：R587.1
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本文編號：2455078