生地黄汤不同配伍比例干预“肾虚血瘀”型崩漏的实验研究

Intervention of Shengdihuang decoction with different compatibilities for uterine bleeding in rats with kidney deficiency and blood stasis

目的观察不同比例生地黄配伍大黄对大鼠"肾虚血瘀"型崩漏的治疗作用,筛选两者配伍比例,初步探讨其配伍规律。方法采用羟基脲+氢化可的松制备大鼠"肾虚血瘀"模型,筛选造模成功的56只大鼠随机分为8组:空白对照组,模型组,宫血宁组,生地黄组,大黄组,生地黄-大黄60∶1组、30∶1组、15∶1组,每组7只。除空白对照组与模型组外,其余各组于造模成功后第2日开始灌胃治疗药物,连续8 d。观察各组大鼠子宫、卵巢体质量及组织病理变化,测定血液流变学、凝血四项相关指标、大鼠血浆中超氧化物歧化酶(SOD1)、过氧化脂质(LPO)、血栓素A2(TXA2)、前列腺素E2(PGE2)、雌激素受体β(ESR2)、肌酐(Cr)、雌二醇(E2)、孕酮(P)含量的变化以及免疫组化法检测对子宫、卵巢雌激素受体蛋白α表达的影响。结果与模型组比较,生地黄-大黄配伍30∶1组可显著延长凝血酶原时间(PT)、活化部分凝血酶原时间(APTT),降低凝血酶时间(TT),降低高、中、低切黏度(P<0.01);显著性升高SOD1、ESR2、TXA2、E2、P(P<0.01),降低LPO、Cr、PGE2(P<0.01);HE染色下观察大鼠卵巢结构显示:30∶1组卵巢发育期卵泡数量较多,多以次级卵泡多见,卵泡细胞、黄体细胞中ER表达明显;30∶1组与模型组比较,子宫内膜均增厚明显,腺体多,基质细胞肥大;ERα在子宫显著表达。结论生地黄-大黄配伍后,对"肾虚血瘀"模型大鼠各项指标改善作用显著,优于单用生地黄及大黄,其最佳配伍比例为生地黄-大黄(30∶1)。止血作用的机制可能与改善血液流变学性能、抗炎、抗氧化及对肾功能、子宫修复等多方面的作用有关。

Objective To observe the therapeutic effect of different compatibilities of rehmanniae combined with rhubarb on uterine bleeding due to kidney deficiency and blood stasis,to screen the proportion of the two and explore the compatibility law.Methods The model of kidney deficiency and blood stasis was prepared by hydroxyurea+hydrocortisone,and 56 rats were randomly divided into 8 groups:a blank control group,a model group,a Gongxuening group,a rehmanniae group,a rhubarb group,3 groups of rehmanniae:rhubard at 60∶1,30∶1,and 15∶1 respectively,7 rats in each group.In addition to the blank control group and the model group,all the other groups were given gavage drugs the second day after the successful modeling for 8 consecutive days.The body mass and histopathological changes of the uterus and the ovary in each group were observed,and the hemorheology and coagulation indexes were determined.The changes of hyper oxide dismutase(SOD1),lipid peroxide(LPO),thromboxane A2(TXA2),prostaglandin E2(PGE2),estrogen receptor 2(ESR2),creatinine(Cr),estradiol(E2)and progesterone(P)contents in the plasma of the rats and the effect of immunohistochemical method on the expression of estrogen receptor protein alpha(ERα)in the uterus and ovary were determined.Results Compared with the model group,rehmanniae and rhizoma compatibility(30:1)group prolonged the prothrombin time(PT),partially activated prothrombin time(APTT),reduced thrombin time(TT),reduced the cut,and lowered the shear viscosity with significant difference(P<0.01);At the same time,SOD1,ESR2,TXA2,E2,and P(P<0.01)were significantly increased,and LPO,Cr,and PGE2 were decreased(P<0.01).The ovarian structure of rats observed by HE staining showed that the 30∶1 group had many follicles in the ovarian development stage,most of which were secondary follicles,with obvious ERαexpression in the follicular cells and luteal cells.Compared with the model group,the endometrium of the 30∶1 group was significantly thickened,with many glands and hypertrophy of the stromal cells.ERαwas well expressed in the utero.Conclusion The compatibility of rehmanniae with rhubarb can significantly improve the indexes of rats with"kidney deficiency and blood stasis",and the effect is better than that of rehmanniae and rhubarb alone.The optimal compatibility ratio is rehmanniae-rhubarb at 30∶1.The mechanism of hemostasis may be related to the improvement of hemorheological properties,anti-inflammation,antioxidation,and affect the renal function and uterine repair.