Xihuang pills induce apoptosis in hepatocellular carcinoma by suppressing phosphoinositide 3-kinase/protein kinase- B/mechanistic target of rapamycin pathway

BACKGROUND The phosphoinositide 3-kinase/protein kinase-B/mechanistic target of rapamycin(PI3K/Akt/mTOR) signalling pathway is crucial for cell survival, differentiation, apoptosis and metabolism. Xihuang pills(XHP) are a traditional Chinese preparation with antitumour properties. They inhibit the growth of breast cancer, glioma, and other tumours by regulating the PI3K/Akt/mTOR signalling pathway. However, the effects and mechanisms of action of XHP in hepatocellular carcinoma(HCC) remain unclear. Regulation of the PI3K/Akt/mTOR signalling pathway effectively inhibits the progression of HCC. However, no study has focused on the XHPassociated PI3K/Akt/mTOR signalling pathway. Therefore, we hypothesized that XHP might play a role in inhibiting HCC through the PI3K/Akt/mTOR signalling pathway.AIM To confirm the effect of XHP on HCC and the possible mechanisms involved.METHODS The chemical constituents and active components of XHP were analysed using ultra-performance liquid chromatography-quadrupole time of flight mass spectrometry(UPLC-Q-TOF-MS). Cellbased experiments and in vivo xenograft tumour experiments were utilized to evaluate the effect of XHP on HCC tumorigenesis. First, SMMC-7721 cells were incubated with different concentrations of XHP(0, 0.3125, 0.625, 1.25, and 2.5 mg/mL) for 12 h, 24 h and 48 h. Cell viability was assessed using the CCK-8 assay, followed by an assessment of cell migration using a wound healing assay.Second, the effect of XHP on the apoptosis of SMMC-7721 cells was evaluated. SMMC-7721 cells were stained with fluorescein isothiocyanate and annexin V/propidium iodide. The number of apoptotic cells and cell cycle distribution were measured using flow cytometry. The cleaved protein and mRNA expression levels of caspase-3 and caspase-9 were detected using Western blotting and quantitative reverse-transcription polymerase chain reaction(RT-qPCR), respectively.Third, Western blotting and RT–qPCR were performed to confirm the effects of XHP on the protein and mRNA expression of components of the PI3K/Akt/mTOR signalling pathway.Finally, the effects of XHP on the tumorigenesis of subcutaneous hepatocellular tumours in nude mice were assessed.RESULTS The following 12 compounds were identified in XHP using high-resolution mass spectrometry:Valine, 4-gingerol, myrrhone, ricinoleic acid, glycocholic acid, curzerenone, 11-keto-β-boswellic acid, oleic acid, germacrone, 3-acetyl-9,11-dehydro-β-boswellic acid, 5β-androstane-3,17-dione, and 3-acetyl-11-keto-β-boswellic acid. The cell viability assay results showed that treatment with 0.625mg/mL XHP extract decreased HCC cell viability after 12 h, and the effects were dose-and timedependent. The results of the cell scratch assay showed that the migration of HCC cells was significantly inhibited in a time-dependent manner by the administration of XHP extract(0.625mg/mL). Moreover, XHP significantly inhibited cell migration and resulted in cell cycle arrest and apoptosis. Furthermore, XHP downregulated the PI3K/Akt/mTOR signalling pathway, which activated apoptosis executioner proteins(e.g., caspase-9 and caspase-3). The inhibitory effects of XHP on HCC cell growth were determined in vivo by analysing the tumour xenograft volumes and weights.CONCLUSION XHP inhibited HCC cell growth and migration by stimulating apoptosis via the downregulation of the PI3K/Akt/mTOR signalling pathway, followed by the activation of caspase-9 and caspase-3.Our findings clarified that the antitumour effects of XHP on HCC cells are mediated by the PI3K/Akt/mTOR signalling pathway, revealing that XHP may be a potential complementary therapy for HCC.

银苓消肿丸联合碳酸氢钠对痛风性关节炎患者骨破坏因子及NLRP3、NF-κB信号通路的影响

目的探讨银苓消肿丸联合碳酸氢钠在痛风性关节炎治疗中的效果及可能机制。方法选取2022年7月至2023年7月该院收治的98例痛风性关节炎患者为研究对象,按照随机数字表法分为对照组与研究组,每组49例,对照组采用碳酸氢钠治疗,研究组在碳酸氢钠基础上联合银苓消肿丸治疗,比较两组临床疗效、关节疼痛、肿胀及活动障碍评分[关节压痛疼痛视觉模拟评分(VAS)、肿胀、活动障碍];比较两组骨破坏因子[破骨细胞分化因子(RANKL)、抗酒石酸酸性磷酸酶-5b(TRACP-5b)、β-胶联降解产物(β-CTX)]、炎症因子[白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)]、核转录因子-κB(NF-κB)、Nod样受体蛋白3(NLRP3)水平,以及不良反应(消化不良、头晕、血压升高、肌肉疼痛)发生情况。结果研究组、对照组总有效率分别为95.92%、77.55%,研究组明显高于对照组,差异有统计学意义(χ^(2)=7.184,P=0.007)。治疗后,两组VAS、肿胀、活动障碍评分均降低,且研究组低于对照组,差异均有统计学意义(P<0.05)。治疗后,两组RANKL、TRACP-5b、β-CTX水平均降低,且研究组低于对照组,差异均有统计学意义(P<0.05)。治疗后,两组IL-1β、TNF-α、NF-κB、NLRP3水平均降低,且研究组低于对照组,差异均有统计学意义(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)。结论在痛风性关节炎的治疗中应用银苓消肿丸联合碳酸氢钠能够改善关节症状及功能,降低炎症因子水平,有效调节NLRP3、NF-κB信号通路,临床应用价值较高,值得推广。

Neuroprotective effects of Ginkgo biloba dropping pills in Parkinson's disease

Parkinson's disease(PD) is the second most common neurodegenerative disease in the world;however,it lacks effective and safe treatments. Ginkgo biloba dropping pill(GBDP), a unique Chinese G. biloba leaf extract preparation, exhibits antioxidant and neuroprotective effects and has a potential as an alternative therapy for PD. Thus, the aims of this study were to evaluate the effects of GBDP in in vitro and in vivo PD models and to compare the chemical constituents and pharmacological activities of GBDP and the G. biloba extract EGb 761. Using liquid chromatography tandem-mass spectrometry, 46 GBDP constituents were identified. Principal component analysis identified differences in the chemical profiles of GBDP and EGb 761. A quantitative analysis of 12 constituents showed that GBDP had higher levels of several flavonoids and terpene trilactones than EGb 761, whereas EGb 761 had higher levels of organic acids.Moreover, we found that GBDP prevented 6-hydroxydopamine-induced dopaminergic neuron loss in zebrafish and improved cognitive impairment and neuronal damage in methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced PD mice. Although similar effects were observed after EGb 761 treatment,the neuroprotective effects were greater after GBDP treatment on several endpoints. In addition, in vitro results suggested that the Akt/GSK3β pathway may be involved in the neuroprotective effects of GBDP.These findings demonstrated that GBDP have potential neuroprotective effects in the treatment of PD.

Liqi Huoxue dripping pill protects against myocardial ischemia-reperfusion injury via the PI3K/Akt/GSK-3β signaling pathway in rats

Background:Liqi Huoxue dripping pill(LQHXDP),a traditional Chinese drug for coronary heart disease,has a protective effect on the heart of rats with myocardial ischemia-reperfusion injury(MIRI)in previous studies;however,its mechanism of action remains unclear.The purpose of this study was to investigate the protective mechanism of LQHXDP on MIRI in rats and its relationship with the PI3K/Akt signaling pathway.Methods:In this study,Sprague-Dawley rats were pre-infused with LQHXDP(175 mg/kg/d)for 10 days.PI3K inhibitor LY294002(0.3 mg/kg)was intravenously injected 15 minutes before ischemia.The rat model of MIRI was established by ligating the left anterior descending coronary artery.Subsequently,cardiac hemodynamics,serum myocardial injury markers,inflammatory factors,myocardial infarct size,antioxidant indexes,myocardial histopathology,and phosphorylation levels of key proteins of PI3K/Akt signaling pathway were assessed in rats.Results:LQHXDP was found to improve cardiac hemodynamic indexes,reduce serum creatine kinase MB isoenzyme activity and cardiac troponin and heart-type fatty acid binding protein levels,lower serum interleukin-1 beta,interleukin-6 and tumour necrosis factorαlevels,reduce the myocardial infarct size and enhance the antioxidant capacity of myocardial tissue in MIRI rats.Pathological analysis revealed that LQHXDP attenuated the extent of myocardial injury and protected mitochondria from damage in MIRI rats.Immunoblot analysis revealed that LQHXDP increased the expression levels of p-Akt and p-GSK-3βin MIRI rat cardiomyocytes.PI3K inhibitor LY294002 could impair these effects of LQHXDP.Conclusion:LQHXDP attenuated myocardial injury,attenuated oxidative stress injury and reduced inflammatory response in MIRI rats,and its protective effects were mediated by activating of PI3K/Akt/GSK-3βsignaling pathway.

Effect of Bak Foong Pills on the expression of β-amyloid in rat retina with optic nerve transection

AIM:To investigate the effect of Bak Foong Pills(BFP) on the expression of β-amyloid(Aβ) in rats retina with optic nerve transaction,and its roles and possible mechanisms in protecting optic nerve damage.· METHODS:Seventy-two healthy,Sprague-Dawley,adult rats were randomly assigned to three groups:negative control group(control group),optic nerve transection group(model group) and BFP treatment group(BFP group,100μg/mL) followed by establishing optic nerve transection model.The expression of Aβ was measured at 48 hours by Western-blotting.Moreover,the expressions of Bcl-2,Bax and Caspase-3 mRNA were evaluated at 48 hours by reverse transcriptase polymerase chain reaction(RT-PCR).RESULTS:There were significant differences among the control,model and BFP groups in the expression of Aβ(all P <0.01).Aβ expression was significantly higher in the model and BFP groups than that in the control group(P < 0.01),with a more significant reduction in the BFP group than that in the model group(P <0.01).Moreover,there were also significant differences among the three groups in the expressions of Bcl-2/Bax(Bcl-2:anti-apoptotic;Bax:proapoptotic) and Caspase-3 mRNA(proapoptotic)(all P<0.01).Bcl-2/Bax ratio was significantly lower and Caspase-3 mRNA expression was significantly higher in the model and BFP groups than those in the control group(P <0.01),with a significant growing of Bcl-2/Bax and reduction of Caspase-3 in the BFP group than those in the model group(P<0.01).· CONCLUSION:BFP can down-regulate Aβ expression in retina and may inhibit apoptosis and protect optic nerve by enhancing Bcl-2/Bax ratio and inhibiting Caspase-3 pathway.

蒙药那如-3味丸对普瑞巴林联合神经阻滞治疗神经病理性疼痛效果的影响

目的探究蒙药那如-3味丸对普瑞巴林联合神经阻滞治疗神经病理性疼痛(NP)效果的影响。方法选择2022年10月至2023年9月确诊为NP的疼痛科住院患者41例,男20例,女21例,年龄40~80岁,BMI≥18.5 kg/m^(2)。采用随机数字表法将患者分为两组:蒙药那如-3味丸组(观察组,n=20)和常规治疗组(对照组,n=21)。对照组为常规治疗,口服普瑞巴林胶囊联合超声引导下疼痛区域神经阻滞;观察组在常规治疗的基础上增加蒙药那如-3味丸(2 g/10粒),每晚临睡前口服3~5粒,共2周。记录治疗前1 d、治疗后2周、1个月、2个月数字评分量表(NRS)疼痛评分、简化McGill疼痛问卷(SF-MPQ)评分、匹兹堡睡眠质量指数(PSQI)。分别于治疗前1 d、治疗后2周采用ELISA法检测血清IL-6、IL-8、IL-1β、TNF-α浓度。记录治疗期间不良反应的发生情况,如恶心、呕吐、腹胀、心悸、嗜睡和头晕等。结果与治疗前1 d比较,治疗后2周、1个月、2个月两组NRS疼痛评分、SF-MPQ评分、PSQI均明显降低(P<0.05);治疗后2周两组血清IL-6、IL-1β、TNF-α浓度明显降低(P<0.05)。与对照组比较,治疗后2周、1个月、2个月观察组NRS疼痛评分、SF-MPQ评分、PSQI均明显降低(P<0.05);治疗后2周观察组IL-6、IL-1β、TNF-α浓度明显降低(P<0.05)。两组不良反应发生率差异无统计学意义。结论蒙药那如-3味丸联合常规疗法可有效减轻NP患者的疼痛,改善患者的睡眠质量,且可能具有调节神经炎症作用。

禾肾丸通过调控ANGPTL3治疗阿霉素肾病大鼠蛋白尿及血脂代谢紊乱的实验研究

目的:探讨禾肾丸通过调控血管生成素样蛋白3(ANGPTL3)治疗阿霉素肾病大鼠蛋白尿及血脂代谢紊乱的作用机制。方法:50只雄性SD大鼠随机分为5组,每组10只,空白组:不作特殊处理,仅予以每日清晨蒸馏水灌胃;模型组:造模成功后每日清晨蒸馏水灌胃;禾肾丸组:造模成功后每日清晨以禾肾丸灌胃;对照组:造模成功后每日清晨以泼尼松灌胃;联合用药组:造模成功后每日清晨以泼尼松、禾肾丸灌胃,各组均灌胃8周。密切观察各组大鼠造模及研究期间各种行为学改变,检测和比较各组大鼠造模前1 d及造模后第2、4、6、8周末24 h尿蛋白水平,同时检测和比较各组大鼠血清总胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)等血脂指标及血清ANGPTL3表达水平。分析各组大鼠血清ANGPTL3表达水平与血脂指标及造模后第8周末24 h尿蛋白水平的相关性。结果:禾肾丸组、对照组及联合用药组大鼠造模后第8周各种行为学改变均优于模型组;各组大鼠造模前1 d 24 h尿蛋白水平比较差异无统计学意义(P>0.05),造模后第2、4周模型组、禾肾丸组、对照组及联合用药组大鼠24 h尿蛋白水平均明显高于空白组(P<0.05),而组间比较差异无统计学意义(P>0.05),造模后第6、8周禾肾丸组、对照组大鼠24 h尿蛋白水平明显高于联合用药组和低于模型组(P<0.05),而禾肾丸组与对照组比较差异无统计学意义(P>0.05);模型组、禾肾丸组、对照组及联合用药组大鼠TC、TG、LDL-C等血脂指标及血清ANGPTL3表达水平均明显高于空白组(P<0.05),而HDL均明显低于空白组(P<0.05),且禾肾丸组、对照组大鼠TC、TG、LDL-C及ANGPTL3均低于模型组和高于联合用药组(P<0.05),HDL-C均高于模型组和低于联合用药组(P<0.05),而禾肾丸组与对照组比较差异无统计学意义(P>0.05);各组大鼠血清ANGPTL3表达水平与TC、TG、LDL-C及24 h尿蛋白均呈正相关关系(P<0.05~P<0.01),而与HDL-C呈负相关关系(P<0.05~P<0.01)。结论:禾肾丸可能通过调控ANGPTL3而起到治疗阿霉素肾病大鼠蛋白尿及血脂代谢紊乱的作用。

Erzhi pills ameliorates cognitive dysfunction and alter proteomic profiles of hippocampus in ovariectomized Alzheimer disease model rats induced by D-galactose and Aβ1-40 injection

OBJECTIVE Erzhi pills is a clas⁃sic prescription of Chinese medicine originated from Fu Shou Jing Fang in Ming dynasty,with the effects of nourishing kidney-Yin and hemosta⁃sis,black hair,strengthening muscles and bones.As a classical prescription for nourishing kidney-Yin,Erzhi pills has been used to treat senile dementia in China for many years.Herein,our study aimed to investigate the protective effects of Erzhi pills in rat models of Alzheimer disease(AD)induced by ovariectomy as well as D-galactose and Aβ1-40 injection and to explore its potential mechanism.METHODS The model of AD rats was established by ovariectomy com⁃bined with D-galactose and Aβ1-40 injection.Ovariectomized rats were randomly divided into four groups:model group,estradiol valerate(0.80 mg·kg-1)group,Erzhi pills high(1.50 mg·kg-1)and low(0.75 mg·kg-1)doses group.In addition,rats of sham operation were selected as the sham operated group.Except for the sham oper⁃ated group,rats were injected intraperitoneally with D-galactose(100 mg·kg-1 per day,for 49 d)on the 8th day,then they were given intracerebro⁃ventricular injection of Aβ1-40(10μg per rat,1 g·L-1)on the 36th day,while the corresponding drugs were given by gastrointestinal administra⁃tion on the 22nd day.In our study,Morris water maze test was used to evaluate the learning and memory abilities,while ELISA kit was used to an⁃alyze serum estrogen level.The morphology of hippocampal neuron cells was observed by HE staining,and Nissl staining was utilized to ob⁃serve the Nissl body in cytoplasm.Then,the ex⁃pression of ERβpositive cells and hippocampal Aβ1-40 and p-Tau404 proteins was determined by immunohistochemistry.In order to further explore the molecular mechanism of Erzhi pills preven⁃tion and treatment of AD,proteomics was used to find potential targets and related pathways,Western blotting was used to verify the expres⁃sion of candidate differential protein.According to the results of proteomics in our experiment,Western blotting was used to detect the protein expression of PI3K,Akt,Bcl-2,Bcl-xl,Bad,14-3-3 and GSK3β.RESULTS The escape latency was significantly shortened,the number of crossing platform was increased,the neuron arrange⁃ment was more orderly and with less nuclear pycnosis in rats of Erzhi pills groups compared to the model group.In rats treated with Erzhi pills,the number of neurons,Nissl bodies,the estro⁃gen levels and ERβpositive cells were increased,while the number of Aβ1-40 and p-Tau404 positive cells was significantly decreased.Proteomics found that there were more than one hundred differentially expressed proteins of rats treated with Erzhi pills,which were involved in 48 signal⁃ing pathways.Among these proteins,five of them were involved in the PI3K/Akt signal path⁃way.As the down-stream protein of PI3K/Akt sig⁃naling pathway,the content of 14-3-3 protein was significantly increased.Western blotting analysis showed that the expression of p-GSK3β/GSK3βand Bad was decreased,while that of p-Akt/Akt,p-PI3K/PI3K,14-3-3,Bcl-xl and Bcl-2 was up-regulated in rats from the Erzhi pills groups compared with the model group.CON⁃CLUSION Erzhi pills can improve estrogen levels,alter proteomics expression of the hippocampus and activate PI3K/Akt pathway in AD rats,reduce Aβaggregation,inhibit the hyperphos⁃phorylation of Tau protein,maintain the morphol⁃ogy of hippocampal neurons and decrease the apoptosis of hippocampal neurons,thereby improving the learning and memory abilities of ovariectomized AD rats induced by D-galactose and Aβ1-40 injection.This study may provide an experimental basis for the clinical treatment of Erzhi pills.

Regulatory Effects of Zuogui Pill on Apoptosis of Follicles in Rats Injured by 60Co-γRays Based on PI3K/Akt/m TOR Signaling Pathway

[Objectives]To explore the protective effects of Zuogui Pill on ^(60)Co-γ-ray-induced premature aging of rats based on phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR)signaling pathway.[Methods]Sixty sexually mature female SD rats were irradiated with ^(60)Co-γ-ray(6.0 Gy,LD 40)for 24 h at one time.These rats were randomly divided into model group,Progynova group[0.18(g·kg)/d],Progynova[0.09(g·kg)/d]+Zuogui Pill high dose[23.625(g·kg)/d)]group,Zuogui Pill high dose[23.625(g·kg)/d)]group,Zuogui Pill medium dose[9.45(g·kg)/d)]group and Zuogui Pill low dose[4.725(g·kg)/d]group.The administration(once a day)lasted 21 d.The rat serum[follicle-stimulating hormone(FSH),luteinizing hormone(LH)and estradiol(E_(2))]were detected by Enzyme-linked immunosorbent assay(ELISA).The morphological changes of ovary were observed by hematoxylin-eosin(HE)staining.The apoptosis rate of granulosa cells was detected by terminal deoxynucleotidyl transferase(TdT)-mediated dUTP nick-end labeling(TUNEL).The protein expression of phosphorylated(p)-PI3K,p-Akt,p-mTOR,B-cell lymphoma-2(Bcl-2),and Bcl-2-associated X protein(Bax)in ovarian tissues were detected by Western blot.[Results]Compared with the normal group,the model group showed significant increase in the serum FSH(P<0.01),significant decrease in serum E_(2)(P<0.05),and decrease in the number of early follicles and luteum in the ovary(P<0.01).Besides,the apoptosis rate of granulosa cells increased significantly(P<0.01);the expression of p-PI3K,p-Akt,p-mTOR and Bcl-2 in ovarian tissue decreased significantly,while the expression of Bax increased significantly(P<0.01).Compared with the model group,the number of early follicles in the ovary increased and the apoptosis rate of granulosa cells decreased after intervention in each administration group.In addition,the protein expressions of p-PI3K,p-Akt,p-mTOR and Bcl-2 increased,while the expression of Bax decreased,especially in Progynova+Zuogui Pill high dose group,the differences were statistically significant(P<0.05,P<0.01).[Conclusions]Zuogui Pill may protect the radiation-injured ovary through activating the expression of PI3K/Akt/mTOR protein in ovarian tissue,increasing the amount of Bcl-2 protein and inhibiting the expression of Bax protein.

基于PI3K/AKT信号通路探讨熄风通脑胶囊对自发性高血压大鼠脑缺血再灌注的干预作用

目的:探讨熄风通脑胶囊对自发性高血压大鼠(SHR)脑缺血再灌注损伤脑组织中磷脂酰肌醇3-激酶(PI3K)和丝氨酸-苏氨酸蛋白激酶蛋白(AKT)的影响。方法:采用改良线栓法建立SHR大鼠局灶性脑缺血再灌注损伤模型,采用随机数字表法将66只SHR大鼠随机分为熄风通脑胶囊高剂量组、熄风通脑胶囊中剂量组、熄风通脑胶囊低剂量组、中风回春丸对照组、模型对照组和假手术对照组,每组11只,另取10只WKY大鼠为正常对照组。通过神经行为学评分、脑组织病理HE染色及检测缺血脑组织中p-PI3K、p-AKT蛋白表达水平,探讨熄风通脑胶囊对脑缺血再灌注损伤SHR大鼠的影响。结果:熄风通脑胶囊高剂量组大鼠神经功能缺损评分低于模型对照组,差异有统计学意义(P<0.05)。脑组织病理HE染色显示,与模型对照组比较,各给药组大脑损伤程度都有一定减轻,其中熄风通脑胶囊高剂量组大鼠脑组织损伤改善情况较熄风通脑胶囊低、中剂量组明显。假手术对照组大鼠脑组织中p-PI3K、p-AKT蛋白相对表达量均低于模型对照组,差异均有统计学意义(P<0.05);熄风通脑胶囊中剂量组、熄风通脑胶囊高剂量组、中风回春丸对照组大鼠脑组织中p-PI3K、p-AKT蛋白相对表达量均高于模型对照组,差异均有统计学意义(P<0.05)。结论:熄风通脑胶囊对SHR大鼠脑缺血再灌注损伤具有保护作用,其保护机制可能与激活PI3K/AKT信号通路,上调p-PI3K和p-AKT蛋白表达有关。

银杏酮酯滴丸联合尤瑞克林治疗急性脑梗死的效果及对神经因子、PTX3和miR-199a的影响

目的探讨银杏酮酯滴丸联合尤瑞克林治疗急性脑梗死的效果及对神经因子、PTX3和miR-199a的影响。方法选择2019年8月至2022年7月收治的78例急性脑梗死患者为研究对象,根据入院时间不同将其分为对照组与观察组,各39例。两组患者入院后均给予对症治疗,对照组在此基础上给予常规溶栓加尤瑞克林治疗,观察组在对照组基础上加银杏酮酯滴丸治疗。比较两组的治疗效果。结果观察组的治疗总有效率高于对照组(P<0.05)。治疗后,观察组的髓鞘碱性蛋白(MBP)、神经生长因子(GFAP)水平低于对照组,胶质纤维酸性蛋白(NGF)水平高于对照组(P<0.05)。治疗后,观察组的正五聚蛋白3(PTX3)水平低于对照组,miR-199a相对表达量高于对照组(P<0.05)。结论银杏酮酯滴丸联合尤瑞克林治疗急性脑梗死的效果满意,可有效改善患者的神经因子水平,调节PTX3水平及miR-199a相对表达量。

基于VEGF/PI3K/Akt通路基因表达探讨骨坏死康复丸对激素性股骨头坏死大鼠血管新生的影响

【目的】观察骨坏死康复丸对激素性股骨头坏死(SONFH)大鼠的治疗作用及机制。【方法】将60只大鼠随机分为空白组,模型组,仙灵骨葆胶囊组及骨坏死康复丸低、中、高剂量组,每组10只。除空白组,其他各组大鼠采用脂多糖联合糖皮质激素诱导法建立SONFH模型。干预结束后,分别进行股骨头的血管造影观察骨髓内微血管变化、苏木素-伊红(HE)染色并计算空骨陷窝率、Micro-CT扫描分析、压缩实验,采用实时定量聚合酶链反应(RT-qPCR)法检测全血磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B(Akt)1、血管内皮生长因子(VEGF)、血小板内皮细胞黏附分子1(CD31)基因表达。【结果】与空白组比较,模型组股骨头髓腔内血供差,空骨陷窝率增加(P<0.05),骨密度、骨体积分数显著降低(P<0.05),股骨头最大载荷及弹性模量降低(P<0.05),全血Akt1、PI3K、VEGF、CD31 mRNA表达水平降低(P<0.05);与模型组比较,骨坏死康复丸高剂量组和仙灵骨葆胶囊组股骨头髓腔内血供较好,空骨陷窝率减少(P<0.05),骨密度、骨体积分数、骨小梁数量、骨小梁厚度显著升高(P<0.05)及骨小梁分离度显著减小(P<0.05),股骨头最大载荷及弹性模量升高(P<0.05),全血Akt1、PI3K、VEGF、CD31 mRNA表达水平升高(P<0.05);骨坏死康复丸高剂量组上述各指标与仙灵骨葆胶囊组比较,差异均无统计学意义(P>0.05)。【结论】骨坏死康复丸可改善大鼠激素性股骨头坏死,其机制与促进VEGF/PI3K/Akt通路基因表达,进而促进血管新生有关。

益气明目丸对视网膜色素变性大鼠视网膜Bax、Caspase-3表达的影响

目的观察益气明目丸对视网膜色素变性大鼠视网膜中Bax、Caspase-3表达的影响。方法24只RCS大鼠分为3组,每组8只,雌雄各半,其中,空白组:RCS(rdy+/+,p+/+)大鼠灌胃生理盐水;模型组:RCS(rdy-/-,p-/-)大鼠灌胃生理盐水;益气明目丸组:RCS(rdy-/-,p-/-)大鼠灌胃益气明目丸悬浊溶液。灌胃30 d后,HE染色观察各组视网膜各层结构,免疫组织化学染色法检测各组视网膜组织中Bax、Caspase-3的平均光密度值,Western blot法测定各组视网膜组织中Bax、Caspase-3蛋白的相对表达量。结果HE染色结果显示,益气明目丸组大鼠视网膜厚度明显高于模型组,感光细胞核数目也较模型组增多,外丛状层、外核层、视杆视锥层较模型组结构更清晰。免疫组织化学染色检测结果显示,益气明目丸组Bax、Caspase-3平均光密度值(0.133±0.030、0.069±0.024)均较模型组(0.211±0.036、0.119±0.027)减少(均为P<0.01)。Western blot检测结果显示,益气明目丸组Bax蛋白、Caspase-3蛋白相对表达量(0.374±0.051、0. 628±0.045)均明显低于模型组(0.768±0.061、0.802±0.074),差异均有统计学意义(均为P<0.01)。结论益气明目丸对视网膜色素变性大鼠视网膜的超微结构具有保护作用;益气明目丸通过抑制视网膜上Bax、Caspase-3的表达减轻视网膜感光细胞的凋亡,达到保护视细胞的目的。

失荅刺知丸对脑缺血再灌注大鼠海马神经元形态学和Caspase-3、Caspase-9表达的影响

目的:观察失荅刺知丸对脑缺血再灌注大鼠海马神经元形态学和Caspase-3、Caspase-9表达的影响。方法:130只健康雄性SD大鼠随机分为假手术组、脑缺血再灌注组(模型组)、失荅刺知丸组(实验组)和金纳多组(阳性对照组),其中后三组根据取材时间点又各分为12 h、24 h和72 h共三个亚组,每组13只。采用线栓法制成大鼠大脑中动脉再灌注模型,透射电镜下观察海马区神经元超微结构变化,免疫组织化学法和Western-blot印迹法检测大鼠海马组织凋亡相关基因Caspase-3、Caspase-9蛋白表达的变化。结果:与假手术组比较,再灌注组大鼠海马神经元出现胞核及线粒体损伤,并且Caspase-3、Caspase-9蛋白的表达明显升高(P<0.05,P<0.01);与再灌注组比较,失荅刺知丸组大鼠海马神经元胞核及线粒体损伤明显减轻,而Caspase-3、Caspase-9蛋白的表达明显降低(P<0.05,P<0.01)。结论:失荅刺知丸可减轻脑缺血再灌注大鼠海马神经元的损伤,其作用机制可能与抑制Caspase-3、Caspase-9蛋白表达有关。

RP-HPLC法测定蒙药那如-3丸中乌头类生物碱含量

目的建立测定蒙药那如 3丸中乌头类生物碱含量的RP HPLC法。方法采用C18色谱柱,以甲醇 水 氯仿 三乙胺(体积比6 0∶4 0∶2∶0 1)为流动相,流速0 8mL·min-1,柱温为室温,检测波长2 5 4nm ,进样体积10 μL,外标法计算含量。结果乌头碱、中乌头碱和次乌头碱在0 3～1 5g·L-1内峰面积与质量浓度呈良好线性关系,相关系数r分别为0 9996、0 9998、0 9998,平均回收率分别为10 1 0 %、10 0 9%、10 0 3% ,RSD分别为1 9%、1 2 %、1 4 % (n =9) ;测得那如 3丸中乌头碱、中乌头碱和次乌头碱的质量分数分别为0 0 0 12 6 %、0 0 0 2 78%、0 0 0 335 %。结论RP HPLC法可用于那如 3丸中乌头类生物碱的质量控制。

蒙药那如-3丸对佐剂性关节炎的治疗作用及机制

目的:研究那如-3丸对大鼠佐剂性关节炎大鼠的治疗作用及部分机制。方法:用弗氏完全佐剂诱导大鼠AA模型;足容积法测量继发性足肿胀度;比色法测定脾淋巴细胞增值反应;用ELISA法测定IL-1、TNF-a、PGE2水平;用比色法测定NO、SOD、MDA含量。结果:致炎后第15天,大鼠继发性关节炎出现,致炎后第8天开始预防给予不同剂量的那如-3丸,连续15d,从致炎后第15天起开始各给药组足跖容积和足跖肿胀度明显下降,同时大鼠关节组织的IL-1、TNFa、PGE2、NO的含量明显下降,但那如-3对SOD、MDA含量无明显影响。结论:那如-3丸对AA大鼠继发性炎症具有治疗作用,起作用机制可能与调节机体异常的免疫功能和维持细胞因子网络平衡有关。

高效液相色谱法测定蒙药那如-3丸中没食子酸的含量

目的 :建立蒙成药那如 - 3丸中没食子酸含量的 HPL C法。方法 :反相高效液相色谱法。采用 C1 8色谱柱 ;以甲醇 -0 .0 0 8mol· L- 1 磷酸水溶液 (5∶ 95 ,v∶ v)作为流动相 ;流速为 1.0 m L· min- 1 ;柱温为室温 ;检测波长为 2 80 nm ;进样体积 5 μL;外标法计算含量。结果 :没食子酸在 80～ 4 0 0 μg·ml- 1范围内峰面积与浓度呈良好线性关系 ,其回归方程为 A=1.16 3× 10 4 C- 3.6 5 1× 10 4 ,r=0 .9992 ,其平均回收率为 10 0 .3% ,RSD为 1.2 % (n=9) ;测得那如 - 3丸中没食子酸的含量为 0 .0 7194 %。结论 :方法简便 ,准确 ,提供了那如 -

滋肾降糖丸联合西药治疗气阴两虚证糖尿病肾病(G3aA2期)的临床观察及对Galectin-3的影响

【目的】观察滋肾降糖丸治疗气阴两虚证糖尿病肾病(G3aA2期)患者的疗效及其对血清半乳糖凝集素-3(Galectin-3)水平的影响。【方法】将60例患者随机分为观察组和对照组,每组各30例。对照组给予西医常规治疗,观察组在西医常规治疗基础上加服滋肾降糖丸,疗程1个月。观察2组患者治疗前后中医证候积分及收缩压(SBP)和舒张压(DBP),肾功能指标[血肌酐(Scr)、肾小球滤过率(eGFR)],尿蛋白指标[尿微量白蛋白肌酐比值(UACR)],糖代谢指标[空腹血糖(FPG)、餐后2 h血糖(2hPG)、糖化血红蛋白(HbA1c)、空腹胰岛素(FINS)、胰岛素抵抗指数(HOMA-IR)],脂代谢指标[甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)]和血清Galectin-3水平的变化情况。【结果】(1)治疗后,2组患者的中医证候总积分及SBP和DBP均较治疗前明显下降(P <0.01),且观察组对中医证候总积分及SBP和DBP的下降作用均明显优于对照组(P <0.01或P <0.05)。(2)治疗后,2组患者的Scr、UACR水平均较治疗前明显下降(P <0.01),eGFR水平均治疗前明显升高(P <0.01),且观察组对Scr、UACR水平的下降作用及对eGFR水平的升高作用均明显优于对照组(P <0.01或P <0.05)。(3)治疗后,2组患者的FPG、2hPG、FINS、HbAlc、HOMA-IR等各项糖代谢指标均较治疗前明显下降(P <0.01),且观察组对上述各项糖代谢指标的下降作用均明显优于对照组(P <0.01或P <0.05)。(4)治疗后,2组患者的TG、TC和LDL-C等各项脂代谢指标均较治疗前明显下降(P <0.01),且观察组对上述各项脂代谢指标的下降作用均明显优于对照组(P <0.05)。(5)治疗后,2组患者的血清Galectin-3水平均较治疗前明显下降(P <0.01),且观察组对血清Galectin-3水平的下降作用明显优于对照组(P <0.05)。【结论】滋肾降糖丸联合西医常规治疗糖尿病肾病(G3aA2期)气阴两虚证患者,具有较好疗效,可有效改善症状,降低血压,改善肾功能,降低尿蛋白,改善糖脂代谢,减轻胰岛素抵抗,并可降低血清Galectin-3水平。

醒脑静对脑出血大鼠脑水肿及血肿周围基质金属蛋白酶-9和补体C3的影响

目的探讨醒脑静改善脑出血后脑水肿的作用机制，及其对血肿周围基质金属蛋白酶-9和补体C3的影响。方法72只健康雄性SD大鼠，随机分为假手术组、出血组和醒脑静组。自体血注射法建立脑出血模型，Garcia法评价神经功能；干湿重法检测脑组织含水量；根据伊文蓝浓度评价血-脑屏障通透性；免疫组织化学检测血肿周围脑组织中基质金属蛋白酶-9和补体C3表达水平。结果经醒脑静治疗后，大鼠神经功能评分逐渐增加（均P〈0．05），至手术后72h达峰值水平（P〈0．05），且手术后48和72h神经功能评分均高于出血组（P〈0．05）。手术后72h，出血组大鼠脑组织含水量升高，血-脑屏障通透性增加，基质金属蛋白酶-9和补体C3表达阳性细胞数目增多（均P〈0．05）；经醒脑静治疗后，脑组织含水量降低，血-脑屏障通透性下降，基质金属蛋白酶-9和补体C3表达阳性细胞数目减少，与出血组比较差异有统计学意义（均P〈0．05）。结论醒脑静通过下调血肿周围脑组织中基质金属蛋白酶-9和补体C3表达水平而达到保护血-脑屏障、减轻血管源性脑水肿的作用。

滋阴明目丸对RCS大鼠视网膜Bax及Caspase-3表达的影响

目的观察滋阴明目丸对皇家外科学院(royal college surgeons,RCS)大鼠视网膜Bax及Caspase-3表达的影响。方法实验对象分3组,分别为:空白组、模型组、滋阴明目丸组(每组雌雄各4只共8只)。空白组:RCS(rdy+/+,p+/+)大鼠,灌胃生理盐水;模型组:RCS(rdy-/-,p-/-)大鼠,灌胃生理盐水;滋阴明目丸组:RCS(rdy-/-,p-/-)大鼠,灌胃滋阴明目丸混悬液。灌胃30 d后,免疫组化法检测各组大鼠视网膜组织中Bax及Caspase-3表达,RT-q PCR法及Western Blot法分别测定各组视网膜组织中Bax及Caspase-3 m RNA及蛋白相对表达水平。结果 Bax、Caspase-3蛋白在滋阴明目丸组视网膜中的阳性表达显著低于模型组,但高于空白组。与空白组比较,模型组Bax、Caspase-3平均光密度值均增加,滋阴明目丸组Bax平均光密度值增加(P<0.05或P<0.01);与模型组比较,滋阴明目丸组Bax、Caspase-3平均光密度值均显著减少(P<0.01)。与空白组比较,模型组Bax、Caspase-3 m RNA、蛋白表达均增加,滋阴明目丸组Bax m RNA、蛋白表达均增加(P<0.05或P<0.01);与模型组比较,滋阴明目丸组Bax、Caspase-3 m RNA、蛋白表达均显著减少(P<0.01)。结论 (1)滋阴明目丸RCS(rdy-/-,p-/-)大鼠视网膜的超微结构具有保护作用;(2)滋阴明目丸能抑制RCS(rdy-/-,p-/-)大鼠视网膜上Bax及Caspase-3的表达;(3)滋阴明目丸可能是通过下调视网膜上Bax及Caspase-3的表达,从而减轻视网膜感光细胞的凋亡,达到保护视细胞的目的。