摘要：
目的:探讨冰片、黄芪甲苷(ASTⅣ)和三七总皂苷(PNS)配伍促进脑缺血再灌注后神经修复的作用和Wnt/β-catenin信号机制。方法:采用大鼠局灶性脑缺血再灌注模型,灌胃给药冰片、ASTⅣ、PNS及其配伍药物,神经功能评分和病理形态评价药物对脑组织损伤的影响,免疫组织化学方法检测脑组织巢蛋白(Nestin)、神经元核抗原(NeuN)反映神经发生,Western blot法检测Wnt/β-catenin信号通路相关蛋白表达。结果:与模型组比较,ASTⅣ+PNS配伍组、冰片+ASTⅣ+PNS配伍高、低剂量组可显著降低神经功能评分(P<0.01),冰片+ A S TⅣ+P N S配伍低剂量组降低的效应均强于各药物单用(P<0.01)。病理学检测表明,各用药组细胞损伤明显减轻,A S TⅣ+P N S配伍组效应显著优于单用A S TⅣ组、单用P N S组,冰片+A S TⅣ+P N S配伍低剂量组效应优于各药物单用及AS TⅣ+P NS配伍组(P<0.01,P<0.05)。A S TⅣ+P N S配伍组、冰片+A S TⅣ+P N S配伍低剂量组使Ne s t i n表达增加,冰片+A S TⅣ+P NS配伍低剂量组的作用显著强于各药物单用及AS TⅣ+P N S配伍组(P<0.01,P< 0.0 5)。A S T Ⅳ+ P N S配伍组、冰片+A S T Ⅳ+ P N S配伍低剂量组使N e u N阳性细胞表达增多,药物配伍的作用显著强于单用A S TⅣ与单用冰片组。A S TⅣ+ P N S配伍组、冰片+ A S TⅣ+ P N S配伍低剂量组使Wnt3a、Wnt7b、β-catenin蛋白表达显著增加,糖原合成酶激酶3β(GSK3β)表达下调,冰片+ ASTⅣ+PNS配伍低剂量组升高Wnt3a、Wnt7b蛋白表达的效应强于各药物单用及ASTⅣ+PNS配伍组(P<0.01, P<0.05),ASTⅣ+PNS配伍组和冰片+ASTⅣ+PNS配伍低剂量组降低GSK3β表达和增强β-catenin蛋白表达的作用强于各药物单用(P<0.01)。结论:冰片、ASTⅣ、PNS配伍能够通过促进脑内神经细胞的增殖,修复受损的神经细胞,增强抗脑缺血再灌注损伤的作用,其作用与调控Wnt/β-catenin信号通路有关。
作者机构：
Department of Integrated Traditional Chinese and Western Medicine, Xiangya Hospital, Central South UniversityProvincial Key Laboratory of Diagnostics of Traditional Chinese Medicine, Hunan University of Chinese Medicine
会议名称：
中国中西医结合学会诊断专业委员会第十三次全国学术研讨会
会议时间：
2019-08-17
会议地点：
中国宁夏银川
会议论文集名称：
中国中西医结合学会诊断专业委员会第十三次全国学术研讨会论文集
关键词：
pharmacokinetic;CSGS;Fluoxetine;LC-MS/MS
摘要：
Context Fluoxetine is a classic serotonin reuptake inhibitor, one of the commonly used antidepressants in the clinic. Simultaneously, Chaihu Shugan（CSGS） powder combined with fluoxetine is commonly used to enhance antidepressant effect and reduce side effects, which could make up for the deficiency of antidepressants, while improving emotional and digestive symptoms. Objective The primary objective of this study was to investigate the potential pharmacokinetic effect of CSGS on fluoxetine in the healthy rats. Materials and methods Thirty-two healthy adult male Sprague-Dawley（SD） rats were divided into four cohorts, one of which was administered Fluoxetine（1.8 mg/Kg·d for consecutive 14 d） alone, while the other three groups were pretreated with low, middle and high doses of CSGS in combination with Fluoxetine(Multiple dose group A（CSGS 5.9 g/Kg·d: fluoxetine 1.8 mg/Kg·d for consecutive 14 d）, Multiple dose group B（CSGS 11.8 g/Kg·d: fluoxetine 1.8 mg/Kg·d for consecutive 14 d）, Multiple dose group C（CSGS 23.6 g/Kg·d: fluoxetine 1.8 mg/Kg·d for consecutive 14 d）. In the fifteenth day, Serial blood samples（0.5 ml） were collected from the tail vein into heparinized 1.5 mL polythene tubes before and 0.25, 0.5, 0.75, 1, 2, 4, 6, 8, 10, 12, 24, 36, 48 h after drug administration. To investigate the herb–drug interaction of CSGSon fluoxetine and its metabolite norfluoxetine using pharmacokinetics, a simple rapid accurate and sensitive Liquid Chromatography-Tandem Mass Spectrometry（LC-MS/MS） method was developed using Sulfamethoxazole（SMZ） as the internal standard（IS）. A liquid-liquid extraction method was applied to extract the analyte from biological samples. All parameters were calculated using Drug and Statistics 3.2.8. Results The linear relationship was good with R2>0.99; the peak was good and no endogenous interference was observed at the retention times; The recovery rate was more than 92%, with the RSD%<10%; RSD% of the stability is less than 10%. Compared with Fluoxetine group, the fluoxetine and norfluoxetine AUC（0–∞） of Combined dose group C significantly increased; while the norfluoxetine AUC（0–∞） in Combined dose group A and Combined dose group B decreased.Compared with Fluoxetine group, the CL of Combined dose group C decreased, while taht of Combined dose group A and B increased. Discussion and conclusion The results of the present study showed that the method is efficient, rapid, sensitive, specific, and has high precision, good recovery and good stability. Overall, high dose of CSS can promote the absortion of fluoxetine and metabolism to norfluoxetine; low and medium dose can inhibit the metabolism of fluoxetine to norfluoxetine and reduce the Cmax of fluoxetine and norfluoxetine.
作者机构：
[陈仙蕾; 丁煌; 唐三; 黄小平; 邓常清; 杨筱倩] Molecular Pathology Laboratory, Key Laboratory of Hunan Province for Prevention and Treatment of Integrated Traditional Chinese and Western Medicine on Cardio-Cerebral Diseases, Key Laboratory of Hunan Universities for Cell Biology and Molecular Techniques, Hunan University of Chinese Medicine, Changsha, 410208, China
通讯机构：
[Huang, X.-P.] M;Molecular Pathology Laboratory, China
摘要：
Collagen triple helix repeat containing 1 (CTHRC1) is a gene that has been associated with tumor progression in human prostate cancer (PC). The tumor immune microenvironment has been linked with disease outcome in PC. In the present study, the correlation between CTHRC1 with PC recurrence and the tumor immunological microenvironment was investigated. Using the data supplied by the Tumor Immune Estimation Resource (TIMER), the expression of CTHRC1, programmed cell death protein 1 (PD-1), and programmed cell death 1 ligand 1 (PD-L1) were analyzed. Immunohistochemical staining of CTHRC1, PD-1 and PD-L1 was performed using a tissue microarray construction of prostate adenocarcinoma (PRAD) specimens. In PRAD, an association was reported between the CTHRC1 expression and the disease free survival (DFS) rate (P=0.022). Overexpression of CTHRC1 was correlated with increased levels of PD-1 (R=0.272, P=0.021) and PD-L1 (R=0.298, P=0.016), elevated levels of infiltrating B cells (P=9.51e(-11)), CD4(+) cells (P=1.51e(-11)), macrophages (P=8.25e(-5)), neutrophils (P=2.17e(-9)) and dendritic cells (P=3.13e(-13)). Bioinformatics analysis revealed that CTHRC1 was correlated with the expression levels of matrix metalloproteinase-9, mucin 1 and solute carrier organic anion transporter family member 2B1 genes, which exert an influence in PRAD. The occurrence of this condition is most likely to be associated with regulation of the tumor microenvironment. Taken together, we demonstrated that the prognosis and immunity of PC are closely linked to CTHRC1 upregulation. Furthermore, these results suggest that the immune function of PC may be suppressed by CTHRC1-targeting therapy.
