Antitumor and synergistic effect of Chinese medicine “Bushen huayu jiedu recipe” and chemotherapy on transplanted animal hepatocarcinoma

AIM： To investigate the antitumor and synergistic effect of Chinese medicine “Bushen huayu jiedu recipe” （recipe for invigorating the kidney, removing blood stasis and toxic substances） and chemotherapy on mice hepatocarcinoma. METHODS： Bushen huayu jiedu recipe （BSHYJDR） consisting of Chinese Cassia Bark, Psoralea, Zedoary, Rhubarb, etc. is equal to 1.5 g/mL liquid of originated herbs after being decoded, filtered, and concentrated. Kunming mice, weighing 18-22 g, were injected with 0.2 mL ascitic hepatocarcinoma H22 containing 1 × 10^7 cells/mL into armpit of the right forelimb of mice. After 24 h, the mice were weighed and randomly divided into tumor-bearing model control group, cisplatin （DDP） group, BSHYJDR high dosage group, low dosage BSHYJDR group, DDP combined with high and low dosage BSHYJDR group, 10 mice in each group. DDP group received injection intraperitoneally （ip） at the dosage of 1 mg/kg （equal to 1/10 LD50）, once a day for 4 d. High and low dosage BSHYJDR groups received intragastric BSHYJDR at the dosages of 26.6 and 13.3 g/kg （20 and 10 times each of clinical adult dosage） respectively, while tumor-bearing model group received the equal volume of distilled water once a day for 10 d. On the 11^th d, the mice were weighed and killed, then the tumor was dissected and weighed, the repression rate （RR） was calculated according to the mean weight of tumor （MWT）. RESULTS： Compared to the model group （MWT： 1.30±0.73）, DDP group （MWT： 0.41±0.09, RR： 68.46%） had a significant difference in the inhibition of hepatocarcinoma H22 （P〈0.01）. High dosage BSHYJDR group （MWT： 0.69±0.29, RR： 46.92%） also had a significant difference in inhibition （P〈0.05）, while no difference was found in low dosage BSHYJDR group （MVVT： 0.85±0.34, RR： 34.62%） （P〉0.05）. When DDP was combined with high dosage BSHYJDR （MWT： 0.29±0.17, RR： 77.69%） and low dosage BSHYJDR （MWT： 0.38±0.21, RR： 70.77%） respectively, we could see improvement of the inhibition effect of DDP on transplanted hepatocarcinoma H22. DDP combined with high dosage BSHYJDR had a significant difference （P〈0.001） compared to DDP, while DDP combined with low dosage BSHYJDR only had a little improvement that is not remarkable. CONCLUSION： Chinese medicine BSHYJDR in combination with chemotherapy can inhibit transplanted hepatocarcinorna in mice.

Effect of Bushen Yiqi Huoxue Recipe on Placental Vasculature in Pregnant Rats with Fetal Growth Restriction Induced by Passive Smoking

Interactions of vascular endothelial growth factor （VEGF） with receptors VEGFR1/Fltl and VEGFR2/Flk1, and those of angiopoietins （Ang-1, Ang-2） with receptor Tie2 play important roles in placental angiogenesis. This study investigated vascular morphology and expression of these angiogenic factors in rat placenta on the day 15, 18, 21 of gestation （D 15, D 18 and D21）. The rats were randomly assigned into 3 groups： normal group, model group [fetal growth restriction （FGR） model], and Bushen Tqi Huoxue （BYHR） recipe treatment group （BYHR group, the pregnant rats with FGR were treated with BYHR recipe）. Morphological analysis indicated that during initial villous formation, fetal nucle- ated erythrocytes （FNEs） appeared in maternal blood sinus （MBS）. Subsequently, FNEs were sur- rounded by endothelial cells to form fetal capillary （FC） and then by trophoblast cells to form villi. As pregnancy proceeded, FC density increased progressively with increasing endothelial identification staining （EIS） in normal and BYHR groups. Whereas, villous formation was suppressed, normal in- crease in FC density was impaired and EIS was weakened in model group. Quantitative PCR analysis showed that VEGF and Flkl mRNA increased over gestation in all groups, indicating that VEGF might play a pivotal role in FC growth during late gestation. VEGF mRNA was increased on D15, while de- creased on D21 in model group as compared with normal group and BYHR group. Immunohistochemi- cally, Ang-2 protein was highly expressed in FNEs, gradually disappeared as villi matured, and decreased over gestation in all groups, indicating that Ang-2 might play a pivotal role in villous formation, which was further supported by decreased Ang-2 mRNA and protein expression in model group on D 15. Ang-1 mRNA, Tie2 mRNA and Ang-1/Ang-2 ratio increased from D15 to D18 in all groups as placenta matured. Ang-1 mRNA, Tie2 mRNA and Ang-1/Ang-2 ratio were decreased on D18 in model group as compared with normal and BYHR groups, indicating delayed maturity of FGR placenta. Alterations in angiogenic factors may result in altered placental vasculature and cause placental insufficiency. BYHR recipe could balance the angiogenic factors to promote the formation and maturation of FGR placental vasculature.

Effect of Bushen Yiqi Hexue Recipe (补肾益气和血方) on Pinopodes Expression on Endometrial Surface in Embryo Implantation Dysfunctional Mice

Objective: To explore the therapeutic mechanism of Bushen Yiqi Hexue recipe (补肾益气血方, BYHR), and observe its effect on pinopodes expression on endometrial surface in mifepristone induced embryo implantation dysfunctional mice. Methods: Pregnant Kunming mice were randomly divided into the normal group, the control group and the treated group. Mice in the treated group were administered with BYHR, and those in the normal group and the control group were administered with normal saline, starting from the very first day of pregnancy (Pd1). On the Pd4, mifepristone (RU486) was subcutaneously injected into mice in the control group and the treated group. Pinopodes expression on endometrial surface at 09:30-10:00 pm on Pd4 (regarded as time point 1, T1) and at 09:30-10:00 am on Pd5 (as time point 2, T2) was determined by scanning electron microscopy. Pregnancy rate and average implanted embryos were observed on Pd7. Results: Pregnancy rate and average implanted embryos in the control group were obviously lower than those in the normal group (all P<0.01), while the two parameters were significantly higher in the treated group (P<0.05, P<0.01, respectively) than those in the control group. In the normal group, abundant developing pinopodes were distributed over the whole endometrial surface in T1, and they were altered to a great deal of fully developed pinopodes in T2. But in the control group, only a few pinopodes were expressed locally on endometrial surface in T1, showing a nonsynchronous figure of development, and they completely disappeared in T2. In the treated group, a lot of developing pinopodes were expressed like those in the normal group but somewhat lagged behind, whereas many fully developed pinopodes were expressed in T2. Conclusion: The decrease of the pinopodes in T1 and the cleaning up of them in T2 is possibly the partial mechanism of mifepristone in inducing embryo implantation dysfunction. It is indicated that BYHR could improve pinopodes expression on endometrial surface, eventually better the uterine receptivity and improve the embryo implantation.

补肾益气化瘀冲剂促进骨质疏松大鼠骨折术后骨痂血管形成的作用机制

背景:补肾益气化瘀冲剂具有益气化瘀、补肾通络的功效,常用于治疗骨质疏松,目前关于补肾益气化瘀冲剂对骨质疏松性骨折骨痂血管形成的影响研究较少。目的:探究补肾益气化瘀冲剂上调细胞外调节蛋白激酶/丝裂原活化蛋白激酶(extracellular signal-regulated kinase/mitogen-activated protein kinases,ERK/MAPK)信号通路改善骨质疏松大鼠骨折术后骨痂血管形成的具体机制。方法:①收集骨质疏松SD大鼠骨髓间充质干细胞、C57BL/6小鼠骨髓单核细胞,MTT法检测不同剂量补肾益气化瘀冲剂对骨髓间充质干细胞的毒性。分别使用添加0,1.5 mg/mL补肾益气化瘀冲剂的培养基培养骨髓间充质干细胞、骨髓单核细胞,进行体外成骨分化、破骨分化实验。②144只SD大鼠随机分为假手术组、模型组、冲剂组、冲剂+PD98059组,每组36只,后3组切除双侧卵巢建立骨质疏松模型,假手术组仅切除卵巢附近部分脂肪组织。8周后所有大鼠接受左侧胫骨横断截骨,冲剂+PD98059组给予5 g/kg补肾益气化瘀冲剂灌胃,尾静脉注射0.3 mg/kg PD98059(ERK1/2抑制剂);冲剂组给予5 g/kg补肾益气化瘀冲剂灌胃,尾静脉注射等体积生理盐水;假手术组、模型组给予等体积生理盐水,尾静脉注射等体积生理盐水;每日给药1次,持续8周。X射线、微型计算机断层扫描、微血管灌注造影观察骨折愈合与骨痂血管生成情况,检测骨痂骨密度与机械强度,苏木精-伊红、Masson染色观察胫骨组织形态,Western blotting、免疫组化检测ERK/MAPK信号通路相关分子表达情况。结果与结论:①与0 mg/mL补肾益气化瘀冲剂组相比,1.5 mg/mL补肾益气化瘀冲剂组成骨细胞碱性磷酸酶活性、矿化水平、p-ERK1/2、p-p38 MAPK表达水平升高(P<0.05),破骨细胞密度、体积减小(P<0.05);②与假手术组相比,冲剂组大鼠第4,8周骨折愈合程度相近,Lane-Sandhu评分、愈伤组织总体积、矿化愈伤组织体积、矿化愈伤组织体积/愈伤组织总体积、骨小梁厚度、血管数量、间距、厚度、血管体积/总体积、骨密度、骨痂极限荷载、刚度、p-ERK1/2、p-p38 MAPK、p-IκB-α、p-p65表达水平差异无显著性意义(P>0.05),模型组、冲剂+PD98059组大鼠骨折愈合缓慢,Lane-Sandhu评分、愈伤组织总体积、矿化愈伤组织体积、矿化愈伤组织体积/愈伤组织总体积、骨小梁厚度、血管数量、厚度、血管体积/总体积、骨密度、骨痂极限荷载、刚度、p-ERK1/2、p-p38 MAPK、p-IκB-α、p-p65表达水平降低(P<0.05),血管间距增加(P<0.05);③提示补肾益气化瘀冲剂能够通过增强ERK/MAPK信号通路相关分子表达改善骨质疏松大鼠骨折愈合情况,促进骨痂血管形成,减轻骨质疏松症状。

补肾益气方对乳腺癌化疗后骨髓抑制及免疫功能影响的临床研究

目的探究补肾益气方对乳腺癌化疗后骨髓抑制及免疫功能的影响。方法选取解放军总医院第六医学中心2023年1月至2024年1月收治的110例经表柔比星+环磷酰胺序贯多西紫杉醇(EC-T)方案化疗的乳腺癌患者作为研究对象,采用随机方法分为对照组和观察组,各55例。其中对照组给予EC-T方案治疗,观察组在对照组基础上口服补肾益气方治疗。观察2组患者治疗前后血常规指标、细胞免疫功能、中医症状积分、生存质量评分(QLQ-C30)变化。结果治疗后,2组白细胞计数、中性粒细胞绝对值、血红蛋白、血小板计数水平均低于治疗前,但观察组均高于对照组(均P<0.05)。治疗后,2组CD_(3)^(+)、CD_(4)^(+)水平及CD_(4)^(+)/CD_(8)^(+)比值均低于治疗前,但观察组均高于对照组,CD_(8)^(+)水平均高于治疗前,但观察组低于对照组(均P<0.05)。治疗后,2组气短乏力、心悸失眠、头晕眼花、面色少华、畏寒肢冷、腰膝酸软积分均高于治疗前,但观察组均低于对照组(均P<0.05)。治疗7、20 d后,2组QLQ-C30评分均低于治疗前,但观察组均高于对照组[(75.2±2.1)分比(74.1±1.9)分、(78.9±2.2)分比(75.9±2.1)分](均P<0.05)。结论补肾益气方能够有效预防乳腺癌化疗后骨髓抑制情况,纠正化疗后免疫功能紊乱情况,同时亦能改善患者化疗后继发的临床症状,明显提高生存质量,可以作为乳腺癌化疗患者的辅助补充治疗。

益气清利化瘀方治疗原发性IgA肾病

目的探讨益气清利化瘀方对脾肾气虚兼湿瘀证IgA肾病患者症状改善和炎性因子水平的影响。方法选取2020年3月—2021年3月在我院接受治疗的264例脾肾气虚兼湿瘀证原发性IgA肾病患者作为研究对象,依据治疗方法不同分为对照组与治疗组,各132例。对照组予以氯沙坦钾治疗,治疗组在对照组基础上予以益气清利化瘀方治疗。分析益气清利化瘀方对该证型肾病患者中医症状改善状况、肾功能及炎症因子变化情况,以及治疗的有效性和安全性分析。结果治疗后,2组尿红细胞数、24 h-UTP、Scr、BUN、UA水平均降低;与对照组比较,益气清利化瘀方组降低幅度更显著(P<0.05);治疗后,2组中医证候积分均降低,益气清利化瘀方组积分低于对照组(P<0.05);益气清利化瘀方组治疗有效率(95.45%)高于对照组(68.18%),治疗后,2组hs-CRP、TNF-α、IL-2、IL-4、IL-5、IL-6水平降低,IFN-γ、IL-10水平升高;与对照组比较,益气清利化瘀方组变化幅度更显著(P<0.05);组间比较,差异具有统计学意义(P<0.05)。结论益气清利化瘀方对脾肾气虚兼湿瘀证原发性IgA肾病患者的中医症状、尿红细胞计数及肾功能改善作用明显,可降低炎性反应,安全性可靠。

Effect of Yiqi Jianpi Bushen Recipe (益气健脾补肾方) in Reducing Nephrotoxicity of Cisplatin

Objective: To observe the clinical effect of Yiqi Jianpi Bushen recipe (YQJPBS, 益气健脾补肾方) in antagonizing nephrotoxicity and hematological toxicity by cisplatin (CDDP). Methods: The patients with different types of carcinomas were divided into two groups. One group was treated by western medicine (WM), i.e. cisplatin combined with diuretic therapy. The other group by Chinese medicine (CM). One day before chemotherapy, YQJPBS was given orally. Urinary enzyme (NAG, γ-GT, ALP) and serum erythropoietin (Epo) were used as the key monitoring indicator. Results: The early renal damage by CDDP remains near proximal convoluted tubule, which leads to the rising of urinary enzyme. So urinary enzyme of the CM group was obviously lower than that of the WM group. The lesion of renal interstitial cells that produce Epo did not cause the serum Epo of WM group to rise obviously 15 days later. As a result there was 40% patients whose RBC and Hb became lower than the standard count. While serum Epo of CM group was higher than that of WM group, only 5% patients in the CM group had their RBC and Hb lowered. Conclusion: YQJPBS can effectively antagonize nephrotoxicity and hematological toxicity induced by CDDP.

益气养阴化瘀方加减治疗早期糖尿病肾病效果及对患者炎性指标和高凝状态的影响

目的探讨益气养阴化瘀方加减在早期糖尿病肾病(气阴两虚、瘀血阻络型)患者治疗中的应用价值,观察其对患者炎性指标、高凝状态的影响。方法选取2020年1月至2022年8月因早期糖尿病肾病(气阴两虚、瘀血阻络型)于广州中医药大学第一附属医院和广东省深圳市第三人民医院门诊及住院的80例患者,采用随机数字表法分为对照组和观察组,各40例。对照组采用常规西医治疗,观察组在对照组基础上联合益气养阴化瘀方加减口服治疗,2组均治疗12周。比较2组治疗前后转化生长因子β1(TGF-β1)、胱抑素C、白细胞介素18(IL-18)、高敏C反应蛋白(hs-CRP)、24 h尿蛋白、血肌酐、D-二聚体、纤维蛋白原水平和临床疗效。结果2组治疗后TGF-β1、胱抑素C、IL-18、hs-CRP、24 h尿蛋白、血肌酐、D-二聚体、纤维蛋白原水平均低于治疗前,且观察组均低于对照组(均P<0.05)。观察组总有效率高于对照组[95.0%(38/40)比72.5%(29/40)],差异有统计学意义(P=0.006)。结论益气养阴化瘀方加减可以有效治疗早期糖尿病肾病(气阴两虚、瘀血阻络型),减轻炎症反应,改善高凝状态。

补肾益气活血方对FGR大鼠小肠上LAT1表达的影响

目的观察补肾益气活血方对宫内生长受限(fetal growth restriction,FGR)大鼠小肠上氨基酸转运蛋白LAT1表达的影响,探讨该方治疗胎儿宫内生长受限的可能机制。方法应用被动吸烟法构建Wistar大鼠FGR模型,将35只孕鼠随机分为空白组、模型组、中药组、西药组及中西药结合组,其中5只孕鼠未获得胎鼠,最终30只孕鼠入组。采用Realtime RT-PCR法、Western Blot法检测胎鼠及孕鼠小肠上氨基酸转运蛋白LAT1(SLC7A5)及其重链4F2hc(SLC3A2)的mRNA及蛋白表达,并比较其表达差异。结果FGR模型组孕鼠小肠上氨基酸转运蛋白LAT1及其对应mRNA SLC7A5表达上调,经补肾益气活血方和(或)精氨酸治疗后出现不同程度的表达下调,且各治疗组胎鼠体重均高于FGR模型组。结论孕鼠小肠上LAT1蛋白及mRNA表达下调可能与补肾益气活血方治疗宫内生长受限的作用机制有关。

益气化瘀方联合针刺治疗胃癌气虚血瘀证患者的效果

目的:观察益气化瘀方联合针刺治疗胃癌气虚血瘀证患者的效果。方法:选取2019年12月至2021年12月该院收治的104例胃癌气虚血瘀证患者进行前瞻性研究,按照随机数字表法将其分为对照组和观察组各52例。两组均进行化疗,在此基础上,对照组采用针刺治疗,观察组在对照组基础上联合益气化瘀方治疗。比较两组客观缓解率(ORR)、疾病控制率(DCR)、治疗前后T细胞亚群指标(CD4^(+)、CD8^(+)、CD4^(+)/CD8^(+))水平,以及不良反应发生率。结果:两组ORR比较,差异无统计学意义(P>0.05);观察组DCR为84.62%(44/52),高于对照组的65.38%(34/52),差异有统计学意义(P<0.05);治疗后,观察组CD4^(+)和CD4^(+)/CD8^(+)水平均高于对照组,CD8^(+)水平低于对照组,差异有统计学意义(P<0.05);观察组白细胞减少、血小板减少、肝损害、胃肠道反应等不良反应发生率均低于对照组,差异有统计学意义(P<0.05)。结论:在化疗基础上,采用益气化瘀方联合针刺治疗胃癌气虚血瘀证患者可提高DCR,改善T细胞亚群指标水平,降低不良反应发生率,效果优于单纯针刺治疗。

补肾化瘀方对卵巢储备功能低下不孕患者子宫内膜容受性的影响

目的观察补肾化瘀方对卵巢储备功能低下(diminished ovarian reserve,DOR)不孕患者子宫内膜容受性的影响。方法纳入70例DOR不孕患者,同时辨证为肾虚血瘀证,用随机数字法分为对照组35例和观察组35例。对照组给予来曲唑、戊酸雌二醇、地屈孕酮治疗,观察组给予来曲唑、补肾化瘀方、地屈孕酮治疗。以3个月经周期为1个疗程,停药后随访3个月经周期。比较两组患者的性激素水平,HCG注射日两组患者的子宫内膜厚度、类型、血流,以及两组妊娠率。结果(1)治疗前后两组患者性激素、AMH比较后,FSH/LH比值、基础血清FSH值较治疗前降低,T和AMH值较治疗前升高;治疗后组间比较,观察组优于对照组,差异均有统计学意义(P<0.05)。(2)HCG注射日,观察组患者的子宫内膜明显增厚、子宫动脉PI、RI降低、A型内膜例数均优于对照组。(3)观察组患者的妊娠率高于对照组。结论补肾化瘀方能改善DOR不孕患者的子宫内膜容受性,提高临床妊娠率。

低频脉冲电磁场联合中药对膝骨关节炎小鼠MMP-13和Col-X的影响

目的从分子生物学、形态学角度,对比观察低频脉冲电磁场联合中药和单纯中药治疗膝关节骨性关节炎(knee osteoarthritis,KOA)小鼠的效果。方法采用Hulth法建立KOA模型小鼠40只,造模成功后分为空白对照组、中药组、低频脉冲电磁场组、联合组,每组各10只。空白对照组和中药组分别予生理盐水、益气补肾化瘀方水煎剂灌胃,每次1 mL,每日2次;低频脉冲电磁场组予低频脉冲电磁场治疗,每天1次,每次30 min;联合组给予中药灌胃的基础上加用低频脉冲电磁场。连续处理2个月后处死小鼠,取膝关节,采用ELISA、qR-TPCR、免疫组化、番红-固绿染色及Micro-CT观察小鼠关节软骨组织功能和形态变化。结果ELISA检测显示,造模后的小鼠血清中IL-1β和TNF-α含量均显著升高,模型制备成功。qRT-PCR和免疫组化结果显示,中药组、低频脉冲电磁场组、联合组的膝关节组织中基质金属蛋白酶-13(MMP-13)、X型胶原蛋白(Col-X)表达水平,及膝关节软骨组织中MMP-13表达量均低于空白组,且联合组最低,差异均有统计学意义(P<0.05);Micro-CT结果显示,联合组小鼠膝关节骨体积分数与骨小梁厚度均高于中药组,差异有统计学意义(P<0.01),2组间骨小梁分离度与骨体积差异无统计学意义(P>0.05)。结论低频脉冲电磁场联合益气补肾化瘀方可降低KOA小鼠膝关节组织中Col-X与MMP-13的表达水平,改善小鼠关节软骨组织功能和形态。

补肾益气活血方治疗气虚血瘀型轻度认知障碍的临床观察

目的观察补肾益气活血方治疗气虚血瘀型轻度认知障碍(mild cognitive impairment,MCI)的临床疗效。方法选取2022年1月至2022年10月就诊于安徽中医药大学第一附属医院脑病科的气虚血瘀型MCI患者60例为研究对象,采用随机对照的研究方法分为治疗组和对照组,每组各30例。两组在保持对症治疗基础疾病的前提下,给予治疗组补肾益气活血方口服,疗程为90 d。分别于治疗第30天、第60天、第90天,对两组患者的中医证候积分进行比较,同时评估两组治疗前后认知功能量表评分和血清炎性因子水平,并分析认知功能和血清炎性指标的相关性。结果与治疗前比较,治疗后治疗组蒙特利尔认知评估量表(Montreal Cognitive Assessment,MoCA)、简易精神状态评估量表(Mini-mental State Examination,MMSE)评分明显升高(P<0.001),且明显高于对照组(P<0.001);治疗后治疗组血清同型半胱氨酸(homocycteine,HCY)、C反应蛋白(C-reaction protein,CRP)、白细胞介素6(interleukin-6,IL-6)及中性粒细胞/淋巴细胞比值(neutrophil to lymphocyte ratio,NLR)水平明显下降(P<0.001),且明显低于对照组(P<0.05);治疗第30天、第60天、第90天,治疗组中医证候总积分、血瘀证候积分、气虚证候积分较治疗前均明显降低(P<0.05或P<0.01),且明显低于对照组(P<0.05或P<0.01);Pearson相关分析显示,血清HCY、CRP、IL-6、NLR水平与MoCA及MMSE评分呈现负相关性(P<0.05)。结论血清炎性因子的升高与认知功能损害密切相关,补肾益气活血方可显著改善气虚血瘀型MCI患者的中医证候评分,降低机体神经炎症水平,提高认知功能,临床疗效明确。

益气养阴活血化瘀方联合降糖补肾丸治疗糖尿病肾病疗效观察

目的:观察益气养阴活血化瘀方联合降糖补肾丸治疗糖尿病肾病(Diabetic nephropathy,DN)的疗效。方法:84例用随机数字表法分为对照组和研究组各42例。两组用降糖补肾丸治疗,研究组用益气养阴活血化瘀方治疗。结果:总有效率研究组高于对照组(P<0.05),治疗8周后腰膝酸痛、疲倦乏力、面色晦暗、气短懒言等评分研究组低于对照组(P<0.05),治疗8周后Scr、BUN、24 h Upro水平研究组低于对照组(P<0.05),治疗8周后血清hs-CRP、TNF-α、ANGPTL4水平研究组低于对照组而CTRP3水平高于对照组(P<0.05),两组不良反应发生率比较差异无统计学意义(P>0.05)。结论:益气养阴活血化瘀方联合降糖补肾丸治疗DN疗效较好,且安全。

补肾化瘀方药对再生障碍性贫血患者骨髓细胞DNA含量的影响

采用计算机图像分析技术，对３０例再生障碍性贫血（ＡＡ）患者经补肾化瘀方药治疗前后的骨髓有核细胞进行ＤＮＡ含量测试分析，并与３０例健康人的测试结果比较，结果表明：ＡＡ患者治疗前骨髓有核细胞ＤＮＡ平均含量明显低于健康人（Ｐ＜０．０１），治疗后与健康人趋于同一水平。补肾化瘀方药影响ＤＮＡ含量的机理可能与改善骨髓微循环或（和）刺激造血细胞从而促进ＤＮＡ合成代谢有关。

益气化瘀补肾方及拆方影响椎间盘细胞外基质胶原和代谢酶mRNA表达的研究

目的探讨益气化瘀补肾方及拆方影响退变模型大鼠颈椎间盘细胞外基质中Ⅰ型、Ⅲ型、Ⅹ型胶原和相关代谢酶MMPs/TI MP mRNA的表达。方法采用RT-PCR法检测胶原和MMPs/TI MP mRNA表达,凝胶成像系统扫描仪对各条带进行扫描,计算机自动计算出光密度。结果与假手术组比较,模型组Ⅰ型、Ⅲ型、Ⅹ型胶原和MMP-13mRNA表达明显升高(P<0.01);TI MP-1mRNA表达无明显统计学差异(P>0.05)。益气化瘀补肾方及其拆方可明显降低Ⅰ型、Ⅲ型、Ⅹ型胶原和MMP-13mRNA的过度表达(P<0.01或P<0.05)。结论退变椎间盘的Ⅰ型、Ⅲ型、Ⅹ型胶原mRNA表达增高,MMP-13mRNA的高表达和TI MP-1mRNA的低表达,MMPs/TI MP的比例失调,反映了退变椎间盘ECM合成和降解的失调。益气化瘀补肾方及拆方可能通过影响颈椎间盘组织胶原及相关代谢酶mRNA表达而延缓椎间盘的退变。

补肾化瘀祛痰方治疗多囊卵巢综合征的临床研究

目的了解补肾化瘀祛痰方治疗多囊卵巢综合征的临床疗效及可能的作用机制。方法以补肾化瘀祛痰中药为主的水煎内服方及外敷方(必要时联合西药二甲双胍)治疗多囊卵巢综合征46例,连续治疗6个月为1个疗程;观察治疗前后临床症状和体征(肥胖、多毛、痤疮及黑棘皮症评分)、血清激素水平及胰岛素释放试验的变化情况。结果治疗后患者多毛、痤疮、黑棘皮症评分值及肥胖评分值与治疗前比较均明显下降(P<0.01,P<0.05);血清睾酮(T)水平较治疗前显著降低(P<0.05);胰岛素试验中空腹胰岛素值与治疗前比较下降(P<0.01),糖负荷后30min、60min胰岛素值及胰岛素曲线下面积(IAUC)与治疗前比较亦有明显下降(P<0.05)。结论以补肾化瘀祛痰方可以显著改善多囊卵巢综合征患者的症状和体征,改善患者的卵巢功能。

益气化瘀补肾方治疗气虚血瘀肾虚型颈椎病大鼠的机制研究

目的:研究益气化瘀补肾方治疗气虚血瘀肾虚型颈椎病大鼠的作用机制。方法:选择3月龄雌性SPF级SD大鼠30只,随机分为正常组、模型组和益气化瘀补肾方组,每组10只。气虚血瘀肾虚型颈椎病模型通过病、证模型复合而成。益气化瘀补肾方组采用益气化瘀补肾方治疗1个月。通过放射免疫学、组织病理学、免疫组织化学和分子生物学等技术研究益气化瘀补肾方的作用机制。结果:与正常组比较,模型组动物出现精神萎靡、少动,舌质、尾色瘀紫,体质量减轻,子宫及附件质量降低,环磷酸腺苷与环磷酸鸟苷及其比值无明显差异,血黏度、血小板表面α-颗粒膜糖蛋白(alpha-granular mem-brane protein,CD62p)增高,血清雌二醇(estradiol,E2)降低。益气化瘀补肾方组与模型组比较,除子宫及附件质量外,上述指标均有不同程度改善。HE染色显示模型组颈椎间盘高度降低,纤维环出现裂隙,胶原纤维排列紊乱,髓核减少,软骨终板变薄;益气化瘀补肾方组椎间盘高度正常,纤维环排列稍紊乱,髓核结构良好,软骨终板变化不明显。与正常组比较,模型组Ⅱ型胶原表达减少;益气化瘀补肾方组Ⅱ型胶原在纤维环的表达比模型组有所增加。模型组与正常组比较Ⅱ型前胶原基因和基质金属蛋白酶抑制剂-1(tissuein-hibitor of metalloproteinase-1,TIMP-1))基因表达降低(P<0.01),益气化瘀补肾方组与模型组比较,Ⅱ型前胶原基因和TIMP-1基因表达增高,基质金属蛋白酶-13基因表达降低(P<0.05,P<0.01)。结论:益气化瘀补肾方通过调节免疫、凝血和内分泌系统等改善模型动物气虚血瘀肾虚状态,通过调节椎间盘细胞外基质以及相关金属蛋白酶延缓椎间盘退变。

益气化瘀补肾方对大鼠肾虚型颈椎病的作用机制

目的:研究益气化瘀补肾方对大鼠肾虚型颈椎病的作用机制。方法:选择3月龄雌性SPF级SD大鼠30只,随机分为正常对照组、肾虚型颈椎病模型组(简称模型组)和益气化瘀补肾方组,每组10只。除正常对照组不施加任何手术外,采用由动静力失衡大鼠颈椎间盘退变模型和去卵巢肾虚模型复合法建立肾虚型颈椎病模型。手术3个月后,益气化瘀补肾方组大鼠予益气化瘀补肾方灌胃治疗1个月。放射免疫法检测血清雌二醇(estradiol,E_2)和血浆环磷酸腺苷(cyclic adenosine monophosphate, cAMP)、环磷酸鸟苷(cyclic guanosine monophosphate,cGMP)含量;锥板法检测血液流变学;流式细胞仪检测血小板表面α-颗粒膜糖蛋白(alpha-granular membrane protein,CD62p)表达;HE染色观察颈椎间盘组织病理学改变,免疫组织化学检测颈椎间盘Ⅱ型胶原蛋白表达情况,并通过实时荧光定量聚合酶链式反应法检测聚集蛋白聚糖(aggrecan-1,Agcl)、Ⅱ型前胶原基因(typeⅡprocollagen gene,Col2a1)、基质金属蛋白酶抑制剂-1(tissue inhibitor of metalloproteinase-1,TIMP-1)、基质金属蛋白酶-13(matrix metalloproteinase-13,MMP-13)mRNA表达。结果:与模型组比较,益气化瘀补肾方组大鼠血清E_2含量升高(P<0.05);血浆cAMP、cGMP含量有所升高,但差异无统计学意义;全血低切黏度、低切还原黏度和CD62p表达百分率下降(P<0.05或P<0.01);颈椎间盘退变有所改善,Miyamoto评分下降(P<0.05),Ⅱ型胶原表达增加;Agcl、Col2a1和TIMP-1 mRNA表达增加(P<0.05,P<0.01),MMP-13 mRNA表达下降(P<0.01)。结论:益气化瘀补肾方可通过调节免疫系统、代谢系统、凝血系统和内分泌系统等指标,改善肾虚型颈椎病证候指标,并延缓椎间盘退变。

益气化瘀方促进糖尿病难愈创面血管新生的AGEs/RAGE/NF-κB信号通路研究

目的:从AGEs/RAGE/NF-κB信号通路探讨益气化瘀方促进糖尿病难愈创面血管新生的作用机制。方法:32只雄性SD大鼠除正常对照组外,腹腔注射链脲佐菌素(STZ)建立糖尿病模型,随机分为模型组、益气化瘀方组、氨基胍组,灌胃8周后在大鼠背部制造全层皮肤缺损开放性创面,于创面造模后第7天取各组大鼠背部创面肉芽组织标本,通过免疫组化及图像分析法检测创面肉芽组织中AGEs、VEGF、CD34含量,用Western blot法检测创面肉芽组织中RAGE、e NOS蛋白水平和NF-κB活性。结果:模型组与正常对照组比较,创面肉芽组织AGEs含量、RAGE蛋白水平、NF-κB活性均明显上升(P<0.01),e NOS蛋白水平、VEGF及CD34含量均明显下降(P<0.01);益气化瘀方组、氨基胍组与模型组比较,创面肉芽组织AGEs含量、RAGE蛋白水平、NF-κB活性均明显下降(P<0.01,P<0.05),e NOS蛋白水平、VEGF及CD34含量均明显上升(P<0.01,P<0.05);益气化瘀方组创面肉芽组织AGEs含量的下降较氨基胍组更为明显(P<0.05)。结论:益气化瘀方可以减少糖尿病大鼠创面肉芽组织AGEs的蓄积及组织细胞表面RAGE的表达,阻断两者结合所导致的组织细胞内NF-κB被激活,细胞受损和功能紊乱,使组织细胞合成和分泌VEGF、NO增加。益气化瘀方可以通过调节AGEs/RAGE/NF-κB信号通路从而促进糖尿病性难愈创面血管新生,加速创面修复愈合。