基于PTEN研究桃红四物汤治疗产后血瘀证的机制
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摘要
目的观察桃红四物汤对产后血瘀证大鼠子宫第10染色体缺失的碱性磷酸和张力蛋白同源物(phosphatase and tensin homolog deleted in chromosome 10,PTEN)mRNA及其蛋白表达的影响,探讨其祛瘀生新的作用机制。方法采用米非司酮和米索前列醇灌胃法复制产后血瘀证大鼠模型,将模型大鼠分为模型组,阳性对照组(益母草颗粒4.3g/kg),桃红四物汤高剂量(18.0g/kg)、中剂量(9.0g/kg)、低剂量(4.5g/kg)组,另设正常对照组。连续灌胃给药7d后,麻醉大鼠并分离子宫,光镜下观察子宫血瘀状态,分别采用实时PCR和Western blot法检测子宫组织PTEN mRNA及其蛋白表达水平。结果桃红四物汤能改善产后血瘀证大鼠子宫间质的充血、水肿。与正常对照组比较,模型组PTEN mRNA及其蛋白表达水平显著升高(P<0.05);与模型组比较,桃红四物汤高剂量组PTEN mRNA及其蛋白表达水平均显著降低(P<0.05),桃红四物汤中剂量组仅PTEN蛋白表达水平显著降低(P<0.05)。结论桃红四物汤的祛瘀生新作用可能与其下调PTEN的表达有关。
Objective To investigate the effect of Taohong Siwu Decoction on the mRNA and protein expression of phosphatase and tensin homolog deleted on chromosome 10(PTEN)in the uterus of rats with postpartum blood stasis syndrome and its mechanism of action in removing blood stasis and promoting tissue regeneration.Methods Rats were given mifepristone and misoprostol by gavage to establish a model of postpartum blood stasis syndrome and then divided into model group,positive control group(treated with Herba Leonuri Granule at a dose of 4.3 g/kg),and high-,middle-,and low-dose Taohong Siwu Decoction groups(treated with Taohong Siwu Decoction at a dose of 18.0,9.0,and 4.5 g/kg,respectively).A normal control group was also established.After 7 consecutive days of treatment by gavage,the rats were anesthetized,and the uterus was collected to observe the status of blood stasis under a light microscope;real-time PCR and Western blot were used to measure the mRNA and protein expression of PTEN in the uterus.Results Taohong Siwu Decoction improved congestion and edema of the uterus in rats with postpartum blood stasis syndrome.Compared with the normal control group,the model group had significant increases in the mRNA and protein expression of PTEN(P<0.05);compared with the model group,the high-dose Taohong Siwu Decoction group had significant reductions in the mRNA and protein expression of PTEN(P<0.05),and the middle-dose Taohong Siwu Decoction group had a significant reduction in the protein expression of PTEN(P<0.05).Conclusion Taohong Siwu Decoction helps to remove blood stasis and promote tissue regeneration possibly by downregulating the expression of PTEN.
Objective To investigate the effect of Taohong Siwu Decoction on the mRNA and protein expression of phosphatase and tensin homolog deleted on chromosome 10(PTEN)in the uterus of rats with postpartum blood stasis syndrome and its mechanism of action in removing blood stasis and promoting tissue regeneration.Methods Rats were given mifepristone and misoprostol by gavage to establish a model of postpartum blood stasis syndrome and then divided into model group,positive control group(treated with Herba Leonuri Granule at a dose of 4.3 g/kg),and high-,middle-,and low-dose Taohong Siwu Decoction groups(treated with Taohong Siwu Decoction at a dose of 18.0,9.0,and 4.5 g/kg,respectively).A normal control group was also established.After 7 consecutive days of treatment by gavage,the rats were anesthetized,and the uterus was collected to observe the status of blood stasis under a light microscope;real-time PCR and Western blot were used to measure the mRNA and protein expression of PTEN in the uterus.Results Taohong Siwu Decoction improved congestion and edema of the uterus in rats with postpartum blood stasis syndrome.Compared with the normal control group,the model group had significant increases in the mRNA and protein expression of PTEN(P<0.05);compared with the model group,the high-dose Taohong Siwu Decoction group had significant reductions in the mRNA and protein expression of PTEN(P<0.05),and the middle-dose Taohong Siwu Decoction group had a significant reduction in the protein expression of PTEN(P<0.05).Conclusion Taohong Siwu Decoction helps to remove blood stasis and promote tissue regeneration possibly by downregulating the expression of PTEN.
引文
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[13]YEOM C J,ZENG L,GOTO Y,et al.LY6E:a con-ductor of malignant tumor growth through modulationof the PTEN/PI3K/Akt/HIF-1axis[J].Oncotarget,2016,7(40):65837-65848.
[14]杨秀梅.桃红四物汤联合自拟补肾汤治疗人工流产术后月经过少临床体会[J].中医临床研究,2018,10(4):115-116.
[2]韩岚,梁杰,张艳艳,等.桃红四物汤对产后血瘀大鼠血清NO、子宫组织VEGF的表达及对PI3K/Akt信号通路的影响[J].中华中医药杂志,2016,31(5):1625-1629.
[3]李白坤,朱继民,梁杰,等.桃红四物汤对产后血瘀证大鼠Rho/ROCK通路RhoA、p-RhoA及PAI-1蛋白表达的影响[J].安徽中医药大学学报,2014,33(1):60-62.
[4]YANG S,KIM H M.The RhoA-ROCK-PTEN path-way as a molecular switch for anchorage dependent cellbehavior[J].Biomaterials,2012,33(10):2902-2915.
[5]YIN D K,LIU Z Q,YANG Y E,et al.Serum contai-ning Tao-Hong-Si-Wu Decoction induces human endo-thelial cell VEGF production via PI3K/Akt-eNOS sig-naling[J].Evid Based Complement Alternat Med,2013,195158[2013-05-21].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3673321.DOI:10.1155/2013/195158.
[6]ZHANG R,LI G,ZHANG Q,et al.Hirsutine inducesmPTP-dependent apoptosis through ROCK1/PTEN/PI3K/GSK3βpathway in human lung cancer cells[J].Cell Death Dis,2018,9(6):598.
[7]CHEN Y C,YUAN T Y,ZHANG H F,et al.Fasudilevokes vasodilatation of rat mesenteric vascular bed viaCa(2+)channels and Rho/ROCK pathway[J].Eur JPharmacol,2016,788:226-233.
[8]XU X Y,ZHANG J,QI Y H,et al.Linc-ROR pro-motes endometrial cell proliferation by activating thePI3K-Akt pathway[J].Eur Rev Med Pharmacol Sci,2018,22(8):2218-2225.
[9]姚亮,李珊珊,于凡,等.正交试验法优选桃红四物汤醇提工艺[J].江西中医药大学学报,2015,27(3):70-72.
[10]梁杰,尹登科,李白坤,等.桃红四物汤对药物致不完全流产早孕大鼠的血管生成及Ang-1,Ang-2,Tie-2表达的影响[J].中国中药杂志,2013,38(21):3731-3715.
[11]LIVAK K J,SCHMITTGEN T D.Analysis of relativegene expression data using real-time quantitative PCRand the 2(-delta delta C(t))methods[J].Methods,2001,25:402-408.
[12]杨文英,温世奇,李瑞丰,等.加味桃红四物汤辅助泡沫硬化剂介入治疗盆腔淤血综合征的疗效观察[J].甘肃医药,2017,36(1):27-29.
[13]YEOM C J,ZENG L,GOTO Y,et al.LY6E:a con-ductor of malignant tumor growth through modulationof the PTEN/PI3K/Akt/HIF-1axis[J].Oncotarget,2016,7(40):65837-65848.
[14]杨秀梅.桃红四物汤联合自拟补肾汤治疗人工流产术后月经过少临床体会[J].中医临床研究,2018,10(4):115-116.