良方温经汤对寒凝血瘀证大鼠子宫及卵巢糖酵解关键酶表达的影响  

Effect of Liangfang Wenjing Decoction on expression of key glycolytic enzymes in uterus and ovaries of rats with coagulating cold and blood stasis syndrome

作　　者：果金玉 邢易 刘鹏 王迪 成秀梅[1] 王鑫国[1,2,3] 任艳青 GUO Jii-yu;XING Yi;LIU Peng;WANG Di;CHENG Xiu-mei;WAN Xin-guo;REN Yan-qing(Hebei University of Chinese Medicine,Shijiazhuang 050091,China;Hebei Traditional Chinese Medicine Quality Evaluation&Standardization Engineering Research Center,Shjiazhuang 050091,China;Hebei Traditional Chinese Medicine Formula Granule Technology Innovation Center,Shijiazhuang 050091,China)

机构地区：[1]河北中医学院,河北石家庄050091 [2]河北省中药材品质评价与标准化工程研究中心,河北石家庄050091 [3]河北省中药配方颗粒技术创新中心,河北石家庄050091

出　　处：《中国中药杂志》2023年第12期3215-3223,共9页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(82004400);河北省自然科学基金项目(H2020423069);河北中医学院优秀青年教师基础研究计划项目(YQ2019009)。

摘　　要：探讨寒凝血瘀证与糖酵解的关系,观察良方温经汤干预对寒凝血瘀证大鼠子宫及卵巢组织糖酵解关键酶表达的影响。采用冰水浴法制备大鼠寒凝血瘀证候模型,于造模结束后进行证候量化评分,并依据评分结果随机分为模型组,良方温经汤低、中、高剂量组(4.7、9.4、18.8 g·kg^(-1)·d^(-1)),每组10只,另设空白组10只。连续灌胃给药4周后,再次进行证候量化评分;采用激光散斑血流成像技术检测各组大鼠耳廓及子宫微循环变化;苏木精-伊红(hematoxylin-eosin, HE)染色观察各组大鼠子宫及卵巢组织病理形态;采用实时定量聚合酶链反应(real-time quantitative polymerase chain reaction, RT-qPCR)及蛋白质印迹法(Western blot)检测各组大鼠子宫及卵巢组织丙酮酸脱氢酶激酶1(pyruvate dehydrogenase kinase 1,PDK1)、己糖激酶2(hexokinase 2,HK2)、乳酸脱氢酶A(lactate dehydrogenase A,LDHA)mRNA与蛋白表达情况。模型组大鼠出现蜷缩少动、舌下脉络增粗、耳廓及子宫微循环血流灌注量降低等寒凝血瘀证候表现,HE染色观察到大鼠子宫内膜变薄,上皮细胞排列混乱,卵巢中卵泡数量减少;与模型组相比,治疗组大鼠寒凝血瘀证候明显缓解,表现为舌质变红、爪甲肿胀减少、尾端未见瘀血,耳部及子宫微循环血流灌注量显著增加(P<0.05或P<0.01),尤以良方温经汤中、高剂量组大鼠改善寒凝血瘀证候最为明显,子宫可见排列整齐的柱状上皮细胞,卵巢中较模型组卵泡数量增多,多见成熟卵泡。与空白组相比,模型组大鼠卵巢及子宫组织中PDK1、HK2、LDHA mRNA与蛋白表达水平显著升高(P<0.05或P<0.01)。与模型组相比,良方温经汤中、高剂量组大鼠卵巢及子宫组织中PDK1、HK2、LDHA mRNA与蛋白表达水平显著降低(P<0.05或P<0.01);良方温经汤低剂量组大鼠子宫及卵巢组织中PDK1、HK2、LDHA mRNA的表达,子宫组织中HK2、LDHA蛋白的表达以及卵巢组织中HK2、PDK1�This study aimed to investigate the relationship between coagulating cold and blood stasis syndrome and glycolysis,and observe the intervention ffeet of Liangfang w enjing Decoction(LFWJD)on the expression of key glycolytie enzymes in the uterus and ovaries of rats with coagulating cold and blood stasis.The rat model of coagulating cold and blood stasis syndrome was established by ice-water bath.After modeling,the quantitative scoring of symptoms were performed,and according to the scoring results,the rats were randomly divided into a model group and LFWJD low-,medium-and high-dose groups(4.7,9.4,18.8 g·kg'.d-'),with 10 in each group.Another 10 rats were selected as the blank group.After 4 weeks of continuous administration by gavage,the quantitative scoring of symptoms was repeated.Laser speckle flowgraphy was used to detect the changes of microcirculation in the ears and uterus of rats in each group.Hematoxylin-eosin(HE)staining was used to observe the pathological morphology of uterus and ovaries of rats in each group.The mRNA and protein expressions of pyruvate dehydrogenase kinase 1(PDK1),hexokinase 2(HK2)and lactate dehydrogenase A(LDHA)in the uterus and ovaries of rats were examined by real-time quantitative polymerase chain reaction(RTqPCR)and Western blot,respectively.The rats in the model group showed signs of coagulating cold and blood stasis syndrome,such as curl-up,less movement,thickened veins under the tongue,and reduced blood perfusion in the microcirculation of the ears and uterus,and HE staining revealed a thinning of the endometrium with disorganized arrangement of epithelial cells and a decrease in the number of ovarian follicles.Compared with the model group,the treatment groups had alleviated coagulating cold and blood stasis,which was manifested as red tongue,reduced nail swelling,no blood stasis at the tail end as well as increased blood perfusion of the microcirculation in the ears and uterus(P<0.05 or P<0.01).Among the groups,the LFWJD medium-and high-dose groups had the most significan

关 键 词：寒凝血瘀证 良方温经汤 糖酵解 丙酮酸脱氢酶激酶1(PDK1) 己糖激酶2(HK2) 乳酸脱氢酶A(LDHA) 

分 类 号：R285.5[医药卫生—中药学]