桃红四物汤调节不完全流产大鼠子宫组织氧化应激诱导的细胞凋亡作用研究
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摘要
目的探讨桃红四物汤是否可以调节不完全流产大鼠子宫组织中由氧化应激诱导的细胞凋亡,发挥对子宫组织的保护作用。方法采用联合米非司酮、米索前列醇复制不完全流产大鼠模型。大鼠随机分为空白对照组、模型组、桃红四物汤高剂量组、桃红四物汤中剂量组、桃红四物汤低剂量组和益母草颗粒阳性对照组。造模及给药期间收集出血量和出血时间数据,给药7 d后进行子宫组织HE染色观察病理状态,测定血清ROS、SOD、GSH、GSH-Px含量,并对组织中Nrf2、HO-1、γ-GCS、Bcl-2、Bax的蛋白表达定量,Tunel染色观察大鼠子宫组织细胞凋亡情况。结果与空白对照组相比,模型组大鼠子宫内膜损伤明显并伴随炎症细胞浸润,ROS明显升高,SOD、GSH、GSH-Px含量较低,Bcl-2蛋白表达量较少,Bax蛋白表达量较高,细胞凋亡较多。与模型组相比,桃红四物汤各剂量组和益母草颗粒组子宫内膜修复明显,ROS降低,SOD、GSH、GSH-Px含量升高,Bcl-2蛋白表达量升高,Bax蛋白表达量降低,细胞凋亡较少。结论桃红四物汤能够显著改善不完全流产大鼠子宫内膜恢复,能降低氧化应激水平及氧化应激介导的细胞凋亡,并很有可能是通过Nrf2通路进行调节。
Objective To determine whether Taohong Siwu decoction may regulate the apoptosis induced by oxidative stress in the uterine tissue of rats with incomplete abortion and protect the uterine tissue. Methods Incomplete abortion rat model was replicated by combining mifepristone and misoprostol. Rats were randomly divided into a blank control group, a model group, Taohong Siwu decoction high-dose, medium-dose group, low-dose groups,and leonurus granule positive control group. Blood loss and bleeding time were collected during the treatment and administration. After 7 days of administration, HE staining of the uterus tissue was performed to observe the pathological state. Then the serum ROS, SOD, GSH, and GSH-Px levels were determined. Nrf2 and HO-1 in the tissue also were observed. The protein expression of γ-GCS, Bcl-2 and Bax was quantified. The apoptosis of rat uterus tissue was determined by Tunel staining. Results Compared with that of the blank control group, the endometrial injury of the model group was accompanied by inflammatory cell infiltration, the ROS was increased,and the SOD, GSH and GSH-Px levels were lowered. The expression of Bcl-2 protein was low, the expression of Bax protein was high, and apoptosis was obvious. Compared with that of the model group, the endometrial repair of Taohong Siwu decoction group and the leonurus granule group was obvious, the ROS was decreased, and the SOD,GSH and GSH-Px levels increased. The expression of Bcl-2 protein was increased, the expression of Bax protein decreased, and apoptosis is was not obvious. Conclusion Taohong Siwu decoction can obviously improve the recovery of the endometrium in rats with incomplete abortion. and reduce the level of oxidative stress and oxidative stress-mediated apoptosis. which is likely to be regulated by Nrf2 pathway.
Objective To determine whether Taohong Siwu decoction may regulate the apoptosis induced by oxidative stress in the uterine tissue of rats with incomplete abortion and protect the uterine tissue. Methods Incomplete abortion rat model was replicated by combining mifepristone and misoprostol. Rats were randomly divided into a blank control group, a model group, Taohong Siwu decoction high-dose, medium-dose group, low-dose groups,and leonurus granule positive control group. Blood loss and bleeding time were collected during the treatment and administration. After 7 days of administration, HE staining of the uterus tissue was performed to observe the pathological state. Then the serum ROS, SOD, GSH, and GSH-Px levels were determined. Nrf2 and HO-1 in the tissue also were observed. The protein expression of γ-GCS, Bcl-2 and Bax was quantified. The apoptosis of rat uterus tissue was determined by Tunel staining. Results Compared with that of the blank control group, the endometrial injury of the model group was accompanied by inflammatory cell infiltration, the ROS was increased,and the SOD, GSH and GSH-Px levels were lowered. The expression of Bcl-2 protein was low, the expression of Bax protein was high, and apoptosis was obvious. Compared with that of the model group, the endometrial repair of Taohong Siwu decoction group and the leonurus granule group was obvious, the ROS was decreased, and the SOD,GSH and GSH-Px levels increased. The expression of Bcl-2 protein was increased, the expression of Bax protein decreased, and apoptosis is was not obvious. Conclusion Taohong Siwu decoction can obviously improve the recovery of the endometrium in rats with incomplete abortion. and reduce the level of oxidative stress and oxidative stress-mediated apoptosis. which is likely to be regulated by Nrf2 pathway.
引文
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[14]段富津.方剂学[M].上海:上海科技出版社,1994:123.
[15]李蕾.益母草颗粒在药物流产中的作用观察[J].实用中医药杂志,2012,28(3):180.
[16]Zhang MJ,An CR,Gao YQ,et al. Emerging roles of Nrf2 and phaseⅡantioxidant enzymes in neuroprotection[J]. Prog Neurobiol,2013,100:30-47.
[17]XuX,ZhangL,YeX,etal. AREpathwayinhibits ROS-induced NLRP3 inflammasome activation in BV2cells after cerebral ischemia reperfusion[J]. Inflamm Res,2018,67(1):57-65.
[18]胡跃强,李媛媛,唐农.大鼠脑缺血再灌注损伤后Nrf2表达变化及清热化瘀方的干预效应[J].中华中医药学刊,2018,36(11):2576-2578.
[19]Chiu AV,Saigh MA,McCulloch CA,et al. The role of NrF2 in the regulation of periodontal health and disease[J].J Dent Res,2017,96(9):975-983.
[20]吴佳怡.细胞凋亡研究进展[J].当代化工研究,2018,(12):190-192.
[21]Yasui K,Baba A. Therapeutic potential of superoxide dismutase(SOD)for resolution of inflammation[J]. Inflamm Res,2006,55(9):359-363.
[22]Dias IH,Chapple IL,Milward M,et al. Sulforaphane restores cellular glutathione levels and reduces chronic periodontitis neutrophil hyperactivity in vitro[J]. PLoS One,2013,8(6):e66407.
[23]邝佩霞,卜莺莺.异甘草素抗肿瘤作用及促凋亡机制研究[J].中南药学,2015,13(8):806-810.
[24]胡戈,曹建民,周海涛,等.虾青素介导Nrf2信号通路减轻大强度运动诱导大鼠心肌细胞的凋亡[J/OL].食品工业科技,2018.
[25]吴灿,王蓉.银杏内酯B对大鼠肝纤维化的防治作用及Nrf2/HO-1和Bcl-2/Bax途径的影响[J].中国新药杂志,2018,27(22):2686-2692.
[2]Tang O,Ho PC. The use of misoprostol for early pregnancy failure[J]. Curr Opin Obstet Gynecol,2006,18(6):581-586.
[3]IyengarK,Klingberg-AllvinM,IyengarSD,etal.Home use of misoprostol for early medical abortion in a low resource setting:Secondary analysis of a randomized controlled trial[J]. Acta Obstet Gynecol Scand,2016,95(2):173-181.
[4]Harlev A,Gupta S,Agarwal A. Targeting oxidative stress to treat endometriosis[J]. Expert Opin Ther Targets,2015,19(11):1447-1464.
[5]褚启龙.氧化应激与细胞凋亡关系的研究进展[J].卫生研究,2003,32(3):276-278.
[6]Zhang Y,Wang J,Zuo C,et al. Protective effect of taohong siwu decoction on abnormal uterine bleeding induced by incomplete medical abortion in rats during early pregnancy[J].Chem Pharm Bull(Tokyo),2018,66(7):708-713.
[7]李白坤,梁杰,朱继民,等.桃红四物汤对产后血瘀证大鼠血清溶血磷脂酸的影响[J].安徽中医学院学报,2013,32(1):50-52.
[8]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):3731-3735.
[9]韩岚,许钒,章小兵,等.桃红四物汤活血化瘀作用的实验研究[J].安徽中医学院学报,2007,26(1):36-38.
[10]李白坤.基于Rho/Rocks通路研究桃红四物汤对产后病血瘀证大鼠活血祛瘀的作用机制[D].合肥:安徽中医药大学,2013.
[11]Li X,Yuan FL,Zhao YQ,et al. Effect of leonurine hydrochloride on endothelin and the endothelin receptor-mediated signal pathway in medically-induced incomplete abortion in rats[J]. Eur J Obstet Gynecol Reprod Biol,2013,169(2):299-303.
[12]Li X,Yuan FL,Zhao YQ,et al. Effects of leonurine hydrochloride on medically induced incomplete abortion in early pregnancy rats[J]. Eur J Obstet Gynecol Reprod Biol,2011,159(2):375-380.
[13]王晓东,赵军宁,张白嘉,等.药物致早孕大鼠子宫出血模型的建立[J].中国药理学通报,1999,15(2):90-92.
[14]段富津.方剂学[M].上海:上海科技出版社,1994:123.
[15]李蕾.益母草颗粒在药物流产中的作用观察[J].实用中医药杂志,2012,28(3):180.
[16]Zhang MJ,An CR,Gao YQ,et al. Emerging roles of Nrf2 and phaseⅡantioxidant enzymes in neuroprotection[J]. Prog Neurobiol,2013,100:30-47.
[17]XuX,ZhangL,YeX,etal. AREpathwayinhibits ROS-induced NLRP3 inflammasome activation in BV2cells after cerebral ischemia reperfusion[J]. Inflamm Res,2018,67(1):57-65.
[18]胡跃强,李媛媛,唐农.大鼠脑缺血再灌注损伤后Nrf2表达变化及清热化瘀方的干预效应[J].中华中医药学刊,2018,36(11):2576-2578.
[19]Chiu AV,Saigh MA,McCulloch CA,et al. The role of NrF2 in the regulation of periodontal health and disease[J].J Dent Res,2017,96(9):975-983.
[20]吴佳怡.细胞凋亡研究进展[J].当代化工研究,2018,(12):190-192.
[21]Yasui K,Baba A. Therapeutic potential of superoxide dismutase(SOD)for resolution of inflammation[J]. Inflamm Res,2006,55(9):359-363.
[22]Dias IH,Chapple IL,Milward M,et al. Sulforaphane restores cellular glutathione levels and reduces chronic periodontitis neutrophil hyperactivity in vitro[J]. PLoS One,2013,8(6):e66407.
[23]邝佩霞,卜莺莺.异甘草素抗肿瘤作用及促凋亡机制研究[J].中南药学,2015,13(8):806-810.
[24]胡戈,曹建民,周海涛,等.虾青素介导Nrf2信号通路减轻大强度运动诱导大鼠心肌细胞的凋亡[J/OL].食品工业科技,2018.
[25]吴灿,王蓉.银杏内酯B对大鼠肝纤维化的防治作用及Nrf2/HO-1和Bcl-2/Bax途径的影响[J].中国新药杂志,2018,27(22):2686-2692.