| 徐新亚,陈碧霞,吴晓婷.基于PI3K/AKT通路探讨何氏补肾厚膜方改善薄型子宫内膜大鼠的作用机制[J].浙江中西医结合杂志,2024,34(3): |
| 基于PI3K/AKT通路探讨何氏补肾厚膜方改善薄型子宫内膜大鼠的作用机制 |
| Exploring the Mechanism of Heshi Bushen Houmo Prescription Improving Thin Endometrium in Rats Based on PI3K/AKT Pathway |
| 投稿时间：2023-09-11 修订日期：2024-03-05 |
| DOI： |
| 中文关键词: 磷脂酰肌醇激酶 蛋白激酶B 何氏补肾厚膜方 薄型子宫内膜 大鼠 |
| 英文关键词:Phosphatidylinositol kinase Protein kinase B Heshi Bushen Houmo Prescription Thin endometrium Rat |
| 基金项目:浙江省基础公益计划项目 |
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| 中文摘要: |
| 目的：基于磷脂酰肌醇激酶（PI3K）/蛋白激酶B（AKT）通路探讨何氏补肾厚膜方（HBHP）改善薄型子宫内膜（TE）大鼠的作用机制。方法：48只SD雌性大鼠，适应1周后按体重随机分为：对照组、模型组、阳性组、HBHP组，每组12只。除对照组外，其他组在发情期对大鼠右侧子宫注入 95%乙醇以建立TE模型。HBHP组在模型建立后灌胃给药HBHP10 g/kg·d，按1 mL/100 g灌胃。阳性组接受小剂量阿司匹林肠溶片0.5 mg/kg·d，按1 mL/100 g灌胃；对照组及模型组每日灌服生理盐水1 mL/100 g。各组大鼠连续灌胃15 d后，剖腹取子宫。苏木精-伊红（HE）用于检测子宫内膜厚度的变化。免疫组织化学（IHC）染色和逆转录-定量实时聚合酶链反应（RT-qPCR）检测子宫内膜组织中PI3K、AKT蛋白和mRNA的表达。结果：与对照组比较，模型组的子宫内膜厚度[(357.34±14.35)μm比(526.08±23.21)μm]、腺体数量[(8.09±1.57)比(19.08±1.31)]、子宫湿重[(0.42±0.05)g比(0.80±0.06)g]和子宫脏器指数[(0.16±0.02)%比(0.30±0.03)%]显著降低（P＜0.05）。与模型组比较，HBHP组、阳性组子宫内膜厚度[(476.74±21.42)μm、(438.18±22.39)μm比(357.34±14.35)μm]、腺体数量[(16.09±1.34)、(13.91±1.45)比((8.09±1.57)]、子宫湿重[(0.75±0.06)g、(0.68±0.08)g比(0.42±0.05)g]和子宫脏器指数[(0.26±0.02)%、(0.24±0.03)%比(0.16±0.02)%]显著增加（P<0.05）。与对照组比较，模型组子宫内膜样本中PI3K、AKT及p-AKT蛋白表达[PI3K:(0.15±0.01)比(0.42±0.02);AKT:(0.21±0.01)比(0.71±0.11);p-AKT:(0.37±0.02)比(0.88±0.06)]和Akt、PI3K mRNA相对表达量[Akt mRNA:(0.42±0.20)比(1.02±0.23);PI3K mRNA:(0.58±0.22)比(1.06±0.35)]显著下调（P<0.05）。与模型组比较，HBHP组、阳性组子宫内膜样本中PI3K、AKT及p-AKT蛋白表达[PI3K:(0.33±0.03)、(0.28±0.02)比(0.15±0.01);AKT:(0.57±0.03)、(0.43±0.05)比(0.21±0.01);p-AKT:(0.71±0.04)、(0.54±0.03)比(0.37±0.02)]和Akt、PI3K mRNA相对表达量[;Akt mRNA:(0.97±0.33)、(0.86±0.30)比(0.42±0.20);PI3K mRNA:(0.98±0.55)、(0.87±0.33)比(0.58±0.22)]显著上调（P<0.05）。结论：HBHP可通过增强PI3K/AKT通路来改善TE大鼠模型的子宫内膜厚度。 |
| 英文摘要: |
| Objective Based on the phosphatidylinositol kinase (PI3K)/Protein kinase B (AKT) pathway to explore the mechanism of Heshi Bushen Houmo Prescription (HBHP) in improving thin endometrium (TE) in rats. Methods 48 SD female rats were randomly divided into: control group,model group,positive group,HBHP group,12 in each group. Except the control group, other groups injected 95% ethanol into the right uterus of rats during estrus to establish TE model. In HBHP group, 10 g/kg·d of HBHP was given by gavage after the model was established, and 1 ml/100 g was given by gavage. The positive group received a small dose of aspirin enteric-coated tablets 0.5 mg/kg·d, 1 ml/100g by gavage; The control group and model group were given normal saline 1 ml/100g daily. Rats in each group were continuously gavaged for 15 d, and then the uterus was removed by laparotomy. HE staining was used to detect the changes of endometrial thickness. Immunohistochemical (IHC) staining and reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) were used to detect the expression of PI3K, AKT protein and mRNA in endometrial tissue. Results Compared with the control group, in the model group,the thickness of endometrium[(357.34±14.35)μm vs (526.08±23.21)μm];the number of glands[(8.09±1.57)vs(19.08±1.31)], the wet weight of uterus[(0.42±0.05)g vs (0.80±0.06)g]and the index of uterine organs[(0.16±0.02)%vs(0.30±0.03)%]
decreased significantly(P＜0.05). Compared with the model group,in the HBHP group and positive group,the thickness of endometrium[(476.74±21.42)μm,(438.18±22.39)μm vs (357.34±14.35)μm], the number of glands[(16.09±1.34),(13.91±1.45)vs((8.09±1.57)], the wet weight of uterus[(0.75±0.06)g, (0.68±0.08)g vs (0.42±0.05)g] and the index of uterine organs[(0.26±0.02)%,(0.24±0.03)%vs(0.16±0.02)%]increased significantly(P＜0.05). Compared with the control group,in the model group,the expression of PI3K, AKT and p-AKT protein[PI3K:(0.15±0.01)vs(0.42±0.02);AKT:(0.21±0.01)vs(0.71±0.11);p-AKT:(0.37±0.02)vs(0.88±0.06)] and the relative expression of Akt and PI3K mRNA in endometrial samples[Akt mRNA:(0.42±0.20)vs(1.02±0.23);PI3K mRNA:(0.58±0.22)vs(1.06±0.35)]decreased significantly(P＜0.05) .Compared with the model group,in the HBHP group and positive group,the expressions of PI3K, AKT and p-AKT proteins[PI3K:(0.33±0.03),(0.28±0.02)vs(0.15±0.01);AKT:(0.57±0.03),(0.43±0.05)vs(0.21±0.01);p-AKT:(0.71±0.04),(0.54±0.03)vs(0.37±0.02)]and the relative expressions of Akt and PI3K mRNA in endometrial samples[Akt mRNA:(0.972±0.33),(0.86±0.30)vs(0.42±0.20);PI3K mRNA:(0.98±0.55),(0.87±0.33)vs(0.58±0.22)]increased significantly(P＜0.05).Conclusions HBHP can improve the endometrial thickness of TE rat by enhancing PI3K/AKT pathway. |
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