| 来坚,李能,刘子彬,张蕊,陈俊卿.当归多糖促进lncRNA MEG3表达调控视网膜上皮细胞焦亡改善糖尿病视网膜病变作用机制研究[J].浙江中西医结合杂志,2023,33(7): |
| 当归多糖促进lncRNA MEG3表达调控视网膜上皮细胞焦亡改善糖尿病视网膜病变作用机制研究 |
| The mechanism of Angelica polysaccharide promoting lncRNA MEG3 expression, regulating retinal epithelial cell pyrosis and improving diabetic retinopathy |
| 投稿时间：2023-03-18 修订日期：2023-06-23 |
| DOI： |
| 中文关键词: 当归多糖 lncRNA MEG3 糖尿病视网膜病变 细胞焦亡 |
| 英文关键词:Angelica polysaccharide lncRNA MEG3 Diabetic retinopathy pyrocytosis |
| 基金项目:当归多糖促进lncRNA MEG3表达调控视网膜上皮细胞焦亡改善糖尿病视网膜病变作用机制研究 浙江省中医药科技计划项目（2022ZA100）） |
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| 中文摘要: |
| 【摘要】目的：探究当归多糖对糖尿病视网膜病变大鼠长链非编码RNA母系表达基因3（lncRNA MEG3）表达调控细胞焦亡的影响及其作用机制。方法：选取糖尿病视网膜病变模型构建成功SD大鼠52只，造模成功当天使用随机数字表法将所有大鼠分为对照组、研究1组、研究2组及抑制组，每组13只，分别给予生理盐水，低剂量当归多糖（15 mg/kg）、高剂量当归多糖（30 mg/kg）和高剂量当归多糖（30 mg/kg）+ lncRNA MEG3抑制剂（2.5 mg/kg）干预，1次/d。观察各组大鼠视网膜病理学改变，定量逆转录PCR和Western blot检测lncRNA MEG3、补丁蛋白（PTCH）、平滑受体蛋白（SMO）、GLI家族锌指蛋白3（GLI3）、蛋白激酶B(PKB) mRNA和蛋白表达，Western blot检测半胱氨酸天冬氨酸蛋白酶-1(Caspase-1)、半胱氨酸天冬氨酸蛋白酶-4(Caspase-4)、半胱氨酸天冬氨酸蛋白酶-5(Caspase-5)蛋白表达，酶联免疫吸附试验（ELISA）检测丙二醛（MDA)、超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）水平。结果：对照组大鼠病理学检测显示视网膜细胞破裂增加，细胞焦亡增多，视网膜神经节细胞结构完整性被破坏，细胞质基质密度降低，细胞核染色质系数，细胞质内形成空泡，研究1组视网膜细胞破裂及细胞焦亡减少，视网膜神经节细胞结构完整性较对照组改善；研究2组神经节细胞结构完整，视网膜细胞破裂、焦亡进一步减少；抑制组在增加lncRNA MEG3抑制剂后逆转了当归多糖作用，显示病例表现与对照组相近。研究1组、研究2组lncRNA MEG3、PTCH mRNA[lncRNA MEG3：（1.59±0.16）、（2.13±0.22）比（1.04±0.11）、（1.02±0.10）；PTCH：（1.41±0.15）、（1.82±0.19）比（1.15±0.13）、（1.17±0.13）；P<0.05]和蛋白[lncRNA MEG3：（1.51±0.15）、（1.99±0.20）比（1.02±0.10）、（0.99±0.10）；PTCH：（1.45±0.16）、（1.81±0.20）比（1.11±0.11）、（1.09±0.10）；P<0.05]相对表达显著高于对照组和抑制组，研究2组显著高于研究1组（P<0.05）；研究1组、研究2组SMO、GLI3、PKB mRNA[SMO：（2.48±0.25）、（2.10±0.22）比（2.94±0.31）、（3.01±0.33）；GLI3：（1.56±0.16）、（1.13±0.11）比（1.95±0.20）、（1.99±0.21）；PKB：（1.43±0.14）、（1.05±0.11）比（1.92±0.19）、（1.91±0.19）；P<0.05]和蛋白[SMO：（2.55±0.26）、（2.07±0.21）比（2.95±0.30）、（2.93±0.29）；GLI3：（1.63±0.17）、（1.14±0.12）比（1.99±0.21）、（2.02±0.23）；PKB：（1.52±0.16）、（1.07±0.11）比（2.04±0.21）、（2.01±0.21）；P<0.05]相对表达显著低于对照组和抑制组，研究2组显著低于研究1组（P<0.05）。研究1组、研究2组MDA水平显著低于对照组和抑制组[（18.03±4.09）、（11.25±3.41）比（25.76±4.34）、（25.32±4.31）；P<0.05]，研究2组显著低于研究1组（P<0.05）；研究1组、研究2组SOD、GSH-Px水平显著高于对照组和抑制组[SOD：（121.48±11.07）、（149.35±12.14）比（94.31±10.13）、（95.12±10.17）；GSH-Px：（131.48±11.62）、（159.07±12.25）比（99.07±10.25）、（100.24±10.31）；P<0.05]，研究2组显著高于研究1组（P<0.05）。研究1组、研究2组显著低于对照组和抑制组[Caspase-1：（3.01±0.32）、（2.49±0.26）比（3.59±0.38）、（3.51±0.36）；Caspase-4：（3.29±0.83）、（2.51±0.71）比（4.13±1.02）、（4.06±1.01）；Caspase-5：（2.70±0.59）、（2.24±0.51）比（3.23±0.68）、（2.17±0.66）；P<0.05]，研究2组显著低于研究1组（P<0.05）。结论：当归多糖能促进糖尿病视网膜病变大鼠lncRNA MEG3表达减少其视网膜上皮细胞焦亡，其作用机制可能与调控SMO/PKB通路从而减少Caspase-1、Caspase-4、Caspase-5蛋白表达有关。 |
| 英文摘要: |
| [Abstract] Objective: To investigate the effects of Angelica polysaccharide on lncRNA MEG3 expression and pyroptosis in diabetic retinopathy rats and its mechanism. Methods: A total of 52 SD rats were selected and diabetic retinopathy models were established. All the rats were divided into control group, study group 1, study group 2 and inhibition group by random number table method. The rats were given normal saline, low-dose angelica polysaccharide（15 mg/kg）, high-dose angelica polysaccharide（30 mg/kg） and high-dose angelica polysaccharide （30 mg/kg）+ lncRNA MEG3 inhibitor（2.5 mg/kg）, once everyday，respectively. Observe the pathological changes in the retina of rats in each group, and detect the mRNA and protein expression of lncRNA MEG3, PTCH, SMO, GLI3, PKB using quantitative reverse transcription PCR and Western blot analysis. Western blot analysis was used to detect Caspase-1, Caspase-4 Caspase-5 protein expression, and the levels of MDA, SOD, and GSH-Px were detected by enzyme-linked immunosorbent assay (ELISA).Retinal pathological changes, lncRNA MEG3, PTCH, SMO, GLI3, PKB mRNA, expression, Caspase-1, Caspase-4, Caspase-5 protein expression, MDA, SOD, GSH-Px levels were observed in each group. Results: Pathological examination of rats in control group showed increased retinal cell rupture, increased pyrosis, damaged structural integrity of retinal ganglion cells, decreased cytoplasmic matrix density, nuclear chromatin coefficient, and formed void in the cytoplasm. Study group 1 showed decreased retinal cell rupture and pyrosis, and improved structural integrity of retinal ganglion cells compared with control group. Ganglion cell structure was intact in the two groups, retinal cell rupture and pyrosis were further reduced. In the inhibition group, the polysaccharide effect of angelica sinensis was reversed after the addition of lncRNA MEG3 inhibitor, indicating that the performance of the cases was similar to that of the control group. Study Group 1, Study Group 2 lncRNA MEG3 PTCH mRNA [lncRNA MEG3: (1.59 ± 0.16), (2.13 ± 0.22) to (1.04 ± 0.11), (1.02 ± 0.10); PTCH: (1.41 ± 0.15), (1.82 ± 0.19) to (1.15 ± 0.13), (1.17 ± 0.13); P<0.05] and protein [lncRNA MEG3: (1.51 ± 0.15), (1.99 ± 0.20) to (1.02 ± 0.10), (0.99 ± 0.10); PTCH: (1.45 ± 0.16), (1.81 ± 0.20) to (1.11 ± 0.10) 1) (1.09 ± 0.10); P<0.05] The relative expression was significantly higher than that of the control group and inhibition group, and the study group 2 was significantly higher than the study group 1 (P<0.05); Study Group 1, Study Group 2 SMO, GLI3 PKB mRNA [SMO: (2.48 ± 0.25), (2.10 ± 0.22) ratio (2.94 ± 0.31), (3.01 ± 0.33); GLI3: (1.56 ± 0.16), (1.13 ± 0.11) ratio (1.95 ± 0.20), (1.99 ± 0.21); PKB: (1.43 ± 0.14), (1.05 ± 0.11) ratio (1.92 ± 0.19), (1.91 ± 0.19); P<0.05] and protein [SMO: (2.55 ± 0.26), (2.07 ± 0.21) ratio (2.95 ± 0.30), (2.93 ± 0.30) 0.29); GLI3: (1.63 ± 0.17), (1.14 ± 0.12) Ratio (1.99 ± 0.21), (2.02 ± 0.23); PKB: (1.52 ± 0.16), (1.07 ± 0.11) ratio (2.04 ± 0.21), (2.01 ± 0.21); P<0.05] The relative expression was significantly lower than that of the control group and the inhibition group, and the study group 2 was significantly lower than the study group 1 (P<0.05). The MDA levels in Study Group 1 and Study Group 2 were significantly lower than those in the control and inhibition groups [(18.03 ± 4.09), (11.25 ± 3.41) compared to (25.76 ± 4.34), (25.32 ± 4.31); P<0.05], and Study Group 2 was significantly lower than Study Group 1 (P<0.05); The levels of SOD and GSH-Px in Study 1 and Study 2 were significantly higher than those in the control and inhibition groups [SOD: (121.48 ± 11.07), (149.35 ± 12.14) compared to (94.31 ± 10.13), (95.12 ± 10.17); GSH-Px: (131.48 ± 11.62), (159.07 ± 12.25) compared to (99.07 ± 10.25), (100.24 ± 10.31); P<0.05], and Study 2 was significantly higher than Study 1 (P<0.05). The levels of Caspase-1: (3.01 ± 0.32), (2.49 ± 0.26) compared to (3.59 ± 0.38), (3.51 ± 0.36); Caspase-4: (3.29 ± 0.83), (2.51 ± 0.71) compared to (4.13 ± 1.02), (4.06 ± 1.01); Caspase-5: (2.70 ± 0.59), (2.24 ± 0.51) compared to (3.23 ± 0.68), (2.17 ± 0.66); P<0.05] were significantly lower in Study 1 and Study 2 compared to Study 1 (P<0.05).Conclusions: Angelica polysaccharide can promote the expression of lncRNA MEG3 in diabetic retinopathy patients and reduce the pyrogen death of retinal epithelial cells, and its mechanism may be related to the reduction of Caspase-1, Caspase-4, Caspase-5 proteins and the protein expressions of PTCH, SMO, CLI3 and PKB. |
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