| 赵峰,綦海燕,肖程程,等.肾气丸对缺血再灌注损伤小鼠肾细胞凋亡的作用及机制研究[J].安徽医药,2020,24(4):656-660. |
| 肾气丸对缺血再灌注损伤小鼠肾细胞凋亡的作用及机制研究 |
| Effect and mechanism of Shen?Qi pills on apoptosis of renal cells in mice with ischemia reperfusion injury |
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| DOI：10.3969/j.issn.1009?6469.2020.04.005 |
| 中文关键词: 急性肾损伤 /中药疗法 再灌注损伤 基因， bcl?2 半胱氨酸天冬氨酸蛋白酶 3 肌酸酐 血尿素氮 超氧化物歧化酶 丙二醛 细胞凋亡 肾气丸 磷脂酰肌醇 3?激酶/蛋白质丝氨酸苏氨酸激酶信号通路 小鼠 |
| 英文关键词: Acute kidney injury/drug therapy(TCD) Reperfusion injury Genes,bcl?2 Caspase 3 Creatinine Blood urea nitrogen Superoxide dismutase Malondialdehyde Apoptosis Shen?Qi pills Phosphatidylinositol?3?kinases(PI3K)/protein?serine?threonine kinase(AKT)signaling pathway Mice |
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| 中文摘要: |
| 目的探讨肾气丸在小鼠肾缺血再灌注( I/R)诱导的急性肾损伤中的作用及机制。方法研究时间为 2018年 4—7月。健康雄性 C57BL6小鼠 40只，利用随机数字表法分为 4组(每组 10只):①假手术组； ②肾缺血再灌注组(肾 I/R组)； ③肾气丸低剂量组； ④肾气丸高剂量组。分组后肾气丸低剂量组每天给予 10.5 g/kg的肾气丸灌胃，肾气丸高剂量组每天给予 21.0 g/kg的肾气丸灌胃，持续 2周；假手术组和肾 I/R组每天给予等量生理盐水灌胃。 2周后构建肾 I/R损伤模型， 24 h处死小鼠并取材。检测血清肌酐( Scr)、尿素氮( BUN)及肾脏组织中丙二醛( MDA)含量和超氧化物歧化酶( SOD)的活性； HE染色及 TUNEL染色检测肾组织损伤及凋亡水平； PCR及免疫组化检测 B淋巴细胞瘤 ?2(Bcl?2)、半胱氨酸天冬氨酸蛋白酶 3(Caspase?3)表达水平；蛋白质印迹法检测 PI3K和 AKT蛋白表达水平。结果 HE及 TUNEL染色表明，与肾 I/R组比较，肾气丸低剂量和高剂量组小鼠肾细胞组织损伤程度减轻，肾小管的上皮细胞肿胀、脱落及凝固坏死等病变损伤减轻，高剂量组较低剂量组损伤程度更轻，明肾气丸可抑制细胞损伤及凋亡。与假手术组中 Scr(135.65±11.21)μmol/L、BUN(8.37±1.80)mmol/L、MDA(3.75±0.33)mmol/L、说SOD(175.69±16.29)U/mL、Caspase?3(0.45±0.06)、 Bcl?2(1.96±0.18)、 PI3K(0.81±0.09)和 AKT(0.86±0.11)蛋白相对表达水平检测指标比较，肾 I/R组 Scr(413.47±36.68)μmol/L、BUN(67.45±8.40)mmol/L、MDA含量( 13.21±1.39)mmol/L、Caspase?3水平(1.76±0.19)均升高( P＜0.05)，SOD活性( 79.36±7.04)U/mL、Bcl?2水平( 0.61±0.09)、 PI3K(0.09±0.01)和 AKT(0.11±0.01)蛋白相对表达水平明显下降( P＜0.05)；与肾 I/R组比较，肾气丸低剂量组 Scr(308.32±17.36)μmol/L、BUN(43.32±4.82)mmol/L和 MDA(11.37±1.02)mmol/L、Caspase?3水平( 1.35±0.16)均降低( P＜0.05)，而 SOD活性( 95.61±9.71)U/mL、Bcl?2(0.94±0.11)、 PI3K(0.23±0.02)和 AKT(0.27±0.03)蛋白相对表达水平明显升高( P＜0.05)；肾气丸高剂量组 Scr(225.59±21.05)μmol/L、BUN(20.63±2.77)mmol/L和 MDA(5.57±1.04)mmol/L、Caspase?3水平( 0.89±0.11)均降低( P＜0.05)，而 SOD活性( 145.72±14.68)U/ mL、Bcl?2(1.52±0.16)、 PI3K(0.45±0.05)和 AKT(0.53±0.06)蛋白相对表达水平也明显升高( P＜0.05)；与肾气丸低剂量组比较，肾气丸组高剂量组 Scr、BUN和 MDA含量、 Caspase?3水平均降低( P＜0.05)SOD活性、 Bcl?2水平、 PI3K和 AKT蛋白表达均升高(P＜0.05)。结论肾气丸可通过激活 PI3K/AKT信号通路对小鼠 I/R诱导的急性肾损伤起保护作用。 |
| 英文摘要: |
| Objective To investigate the protective effect and mechanism of Shen?Qi pills on acute renal injury induced by Isch? emia?reperfusion(I/R)in mice.Methods From April 2018 to July 2018，the total of 40 healthy male C57BL6 micewere randomly assigned into 4 groups:①sham group；②renal ischemia reperfusion group(renal I/R group)； ③low dose group of Shen?Qi pills； ④high dose group of Shen?Qi pills.The low dose group of Shen?Qi pills wasadministered with 10.5 mg/kg/d of Shen?Qi pills，while the high dose group was gavaged with 21.0 mg/kg/d of Shen?Qi pills，for two weeks.The sham and I/R groups were administered the same amount of normal saline every day.After 2 weeks，the model of renal I/R injury was duplicated.The 4 groups were sacrificed after 24h.Then Scr，BUN，the content of MDA and activity of SOD were detected.HE and TUNEL staining were used to detect renalinjury and apoptosis level.The expression levels of bcl?2 and caspase?3 were detected by PCR and Immunohistochemistry.The ex?pression levels of PI3K and AKT were detected by Western blot.Results HE and TUNEL staining showed that compared with the renal I/R group，the damage degree in the Shen?Qi pills group(low dose group and high dose group)was reduced，the damage de? gree of epithelial cells in renal tubules such as swelling，abscission and coagulation necrosis was reduced，and the damage degree in the high dose group of Shen?Qi pills groupwas lighter than that in the low dose group，which demonstrated that the Shen?Qi pillscould inhibit cells damage and apoptosis.Compared with Scr，MDA，SOD，Caspase?3，BUN，Bcl?2，relative expression level of PI3K proteinand AKT protein level in sham group were(135.65±11.21)μmol/L，(8.37±1.80)mmol/L，(3.75±0.33)mmol/L，(175.69±16.29)μ/L，(0.45±0.06)(1.96±0.18)(0.81±0.09)and(0.86±0.11)respectively.Among them，Scr(413.47±36.68)μmol/L，BUN(67.45±8.40)μmol/L， mmol/L and caspase?3 levels1.76±0.19)in the renal I/R group were higher than those in sham group(P＜0.05)and SOD activity(79.36±7.04)U/mL，bcl?2 level(0.61±0.09)，relative expression level of PI3K protein MDA，(13.21±1.39)，((0.09±0.01)andAKTprote，in(0.11±0.01)in the renal I/R group were significantly lower than those in sham group(P＜0.05).Scr(308.32±17.36)μmol/L，BUN(43.32±4.82)mmol/L，MDA(11.37±1.02)mmol/L and caspase?3(1.35±0.16)level in the Shen?Qi pills low dose group were lower than those in the renal I/R group(P＜0.05)，while SOD activity(95.61±9.71)U/mL，Bcl?2 level(0.94±0.11)relative expression level of PI3K protein(0.23±0.02)and AKT protein(0.27±0.03)were increased than those in the renal I/R grouP＜0.05).Scr(225.59±21.05)μmol/L，BUN(20.63±2.77)mmol/L，MDA(5.57±1.04)mmol/L and caspase?3(0.89±0.11)level in the Shen?Qi pills high dose group were decreased than those in the renal I/R group(P＜0.05)，while SOD activity(145.72±14.68)U/mL，Bcl?2 level(1.52±0.16)，relative expression level of PI3K protein(0.45±0.05)and AKT protein(0.53±0.06)were increased than those in the renal I/R group(P＜0.05)； the content of Scr，BUN and MDA and the level of caspase?3 in the high?dose group were decreased than those in the Shen?Qi pills low dose group(P＜0.05)，while the SOD activity，bcl?2 level， PI3K and AKT protein expression level were increased than those in the Shen?Qi pills low dose group(P＜0.05).Conclusion The Shen?Qi pills can protect I/R?induced acute renal injuryof mice by activating the PI3K/AKT signaling pathway. |
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