Mechanism of action of Wuzi Yanzong pill in the treatment of oligoasthenozoospermia in rats determined via serum metabolomics

Objective:To investigate the mechanism of action of Wuzi Yanzong pill(WYP)in rats with oligoasthenozoospermia(OAZ)via metabolomics and to provide a possible basis for improving this WYP-based treatment.Methods:A rat model of OAZ was established by treating male SpragueeDawley rats with glucosides from Tripterygium wilfordii Hook.F.Seventy-two rats were randomly divided into six groups:control,L-carnitine(positive control),model,and low-,medium-,and high-dose WYP groups.Rats in the experimental groups were treated with WYP for 4 weeks.At the end of the treatment period,sperm cell quality(density,motility,and viability)was assessed using a semen analysis system,mitochondrial membrane potential(MMP)was assessed using flow cytometry,and testicular injury was assessed using hematoxylin and eosin staining to validate the therapeutic effect of WYP in OAZ.Further,serum metabolomics-based analysis was performed using high-performance liquid chromatography-mass spectrometry to identify differential metabolic pathways and possible mechanisms of action of WYP in OAZ treatment.Results:A rat model of OAZ was considered successfully-established after comparing the quality of spermatozoa in the model group to that in the control group.WYP-M and WYP-H treatments significantly improved sperm cell density,motility,and viability compared with those in the model group(all P<.05).Compared with the model group,both WYP-M and WYP-H treatments increased MMP values(P=.006 and P=.021 respectively),while there was no significant difference in the L-carnitine group.L-carnitine and WYP administration reversed damage to the testes to varying degrees compared with that in the model group.Further,44 differential metabolites and four metabolic pathways,especially autophagy pathway,related to OAZ were identified via metabolomics.Conclusions:WYP improves sperm cell quality and MMP in OAZ primarily via autophagy regulation.These findings can be employed to improve the efficacy of WYP in humans.

Can Tibetan medicine Honghua Ruyi pills relieve endometriosisassociated dysmenorrhea?Protocol for a randomized placebocontrolled trial

Objective:To provide high-quality clinical evidence of the efficacy of Tibetan medicine Honghua Ruyi(HHRY)pills for endometriosis-associated dysmenorrhea.Methods:This study constitutes a multicenter,randomized,double-blind,placebo-controlled trial encompassing a three-menstrual cycle intervention followed by a three-menstrual cycle follow-up period.A total of 164 eligible females with endometriosis-associated dysmenorrhea were randomly divided into HHRY pills and placebo groups in a 1:1 ratio.The primary outcome included dysmenorrhea symptoms assessed using Visual Analog Scale(VAS)scores and quality of life,whereas the secondary outcome measures included the maximum VAS for non-menstrual pelvic pain,duration of pain episodes(in days),frequency and quantity of the consumption of ibuprofen sustained-release capsules(or other non-steroidal anti-inflammatory drugs),and days off work/study for staff/student due to dysmenorrhea,ovarian cyst,and/or pelvic nodule size.The safety was monitored throughout the treatment period.All the analyses were based on the intention-to-treat principle.For continuous outcomes,simple or multiple linear regressions were used to estimate the differences between the HHRY pills and placebo groups,with categorical data expressed as the number and percentage of occurrences.Differences were compared using the chi-square test or Fisher's exact test.The predefined analysis was adjusted for concomitant treatment,a variable considered to be associated with outcomes but unaffected by treatment allocation.Estimates of treatment effects were reported with 95%confidence intervals.Two-tailed P values≤.05 were considered statistically significant.Conclusion:Positive results from this trial,upon completion would provide robust evidence for the efficacy and safety of HHRY pills in treating dysmenorrhea in patients with endometriosis.

The Effect of Ginkgo Biloba Leaf Dropping Pill Combined with Butylphthalide Capsule on Cognitive Dysfunction in Patients after Acute Ischemic Stroke and Its Impact on Serum Cytokines

Objective: To investigate the therapeutic effect of Ginkgo biloba extract dropping pills combined with butylphthalide capsules on cognitive dysfunction in patients after acute ischemic stroke (AIS) and its impact on serum cytokines CRP, IL-6, and Hcy. Methods: This study selected 76 patients with cognitive dysfunction after ischemic stroke who were hospitalized at Zhuji People’s Hospital from January 2023 to January 2024. The patients were divided into two groups. The control group was treated with butylphthalide capsules, while the combination group received Ginkgo biloba extract dropping pills in addition to the treatment given to the control group. The neurological function, cognitive function, activities of daily living, and levels of serum cytokines CRP, IL-6, and Hcy were compared between the two groups after 1 month and 3 months of treatment. Results: The NIHSS scores, MMSE scores, ADL scores, and levels of CRP, IL-6, and Hcy in both groups showed statistically significant differences compared to before treatment (P Conclusion: The combination of Ginkgo biloba extract dropping pills and butylphthalide capsules has a better therapeutic effect on cognitive dysfunction in patients after ischemic stroke. It can improve the neurological function and cognitive function of patients, enhance their ability to perform daily activities, and reduce inflammatory responses.

Exploring the molecular mechanism of Suoquan pill in the treatment of diabetic kidney disease based on network pharmacology,molecular docking,in vitro experiment

Background:Diabetic kidney disease(DKD)is a microvascular complication of diabetes mellitus and is the main cause of end-stage renal failure.Suoquan pills(SQP)has a variety of pharmacological activities and multiple therapeutic effects,and it is used clinically as a basic formula for the treatment of DKD.Methods:Public databases were used to identify SQP compounds and the potential targets of SQP and DKD.A drug-component-therapeutic target network was constructed.Protein-protein interaction network analysis,Gene Ontology functional analysis,and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were used to analyse the potential molecular mechanisms of SQP based on common targets of drugs and diseases.Molecular docking simulations were conducted to confirm the binding abity of the core compounds to key targets.The efficacy and predicted molecular mechanisms of SQP were validated using cell counting kit-8 assay,flow cytometry,and western blotting with HK-2 cells as a model.Results:Network pharmacology analysis showed that 26 compounds and 207 potential targets of SQP were involved in the treatment of DKD;boldine,denudatin B,pinocembrin,kaempferoid,and quercetin were considered core compounds,and epidermal growth factor receptor(EGFR)and proto-oncogene,non-receptor tyrosine kinase(SRC)were considered key targets.Gene Ontology enrichment analysis indicated that protein phosphorylation and negative regulation of apoptotic processes are important biological processes in the treatment of DKD by SQP.Molecular docking confirmed the excellent binding abilities of boldine,denudatin B,kaempferide,and quercetin to EGFR and SRC.The results of in vitro experiments showed that treatment with an ethanolic extract of SQP significantly protected HK-2 cells from high glucose-induced cell damage.In addition,the SQP ethanol extract inhibited the phosphorylation of EGFR and SRC,suppressed the apoptosis rate,and regulated apoptosis-related proteins in HK-2 cells under high glucose stress.Conclusion:This study systematically and intuitively illustrated the possible pharmacological mechanisms of SQP against DKD through multiple components,targets,and signalling pathways,especially the inhibition of EGFR and SRC phosphorylation and apoptosis.

Integrated network pharmacology and metabolomics to explore the mechanisms of Shenzao dripping pill against chronic myocardial ischemia

Background:Shenzao dripping pill(SZDP)is empirically prescribed for treating cardiac diseases.Nevertheless,there is a lack of comprehensive knowledge regarding the underlying mechanisms contributing to its therapeutic effects.The objective of this study is to investigate the underlying mechanism of SZDP against chronic myocardial ischemia(CMI)in a rat model.Methods:In this study,we utilized electrocardiographic and echocardiographic detection along with pathological tissue analysis to evaluate the efficacy of SZDP.The integration of network pharmacology and metabolomics was conducted to investigate the mechanisms.Molecular docking and molecular dynamics simulations were used to validate the binding energy between the compounds of SZDP and the associated targets.Results:The results showed that SZDP was able to improve T wave voltage,reverse CMI abnormalities in ejection fraction and fractional shortening,and restore histopathological heart damage.Metabolomics results indicated that disturbances of metabolic profile in CMI rats were partly corrected after SZDP administration,mainly affecting purine metabolism.13-Docosenamide may be the potential metabolic biomarker of the therapeutic application of SZDP for CMI.Integrating network pharmacology and metabolomics,thiopurine S-methyltransferase(TPMT),xanthine dehydrogenase/oxidase(XDH),bifunctional purine biosynthesis protein ATIC(ATIC),and cytochrome p4501A1(CYP1A1)were identified as possible targets of SZDP to exert therapeutic effects by enhancing the metabolic levels of L-Tryptophan,Deoxyribose 1-phosphate and Phosphoribosyl formamidocarboxamide.Conclusion:SZDP has a therapeutic effect on CMI by regulating metabolite levels,acting on the targets of TMPT,XDH,ATIC,and CYP1A1,and reducing cardiomyocyte injury and myocardial fibrosis.

Clinical efficacy of Baijin pills in the treatment of generalized tonicclonic seizure epilepsy with cognitive impairment

BACKGROUND The generalized tonic-clonic seizure(GTCS)is the most usual variety of epileptic seizure.It is mainly characterized by strong body muscle rigidity,loss of consciousness,a disorder of plant neurofunction,and significant damage to cognitive function.The effect of antiepileptic drugs on cognition should also be considered.At present,there is no effective treatment for patients with epilepsy,but traditional Chinese medicine has shown a significant effect on chronic disease with fewer harmful side effects and should,therefore,be considered for the therapy means of epilepsy with cognitive dysfunction.AIM To investigate the clinical efficacy of Baijin pills for treating GTCS patients with cognitive impairment.METHODS This prospective study enrolled patients diagnosed with GTCS between January 2020 and December 2023 and separate them into two groups(experimental and control)using random number table method.The control group was treated with sodium valproate,and the experimental group was Baijin pills and sodium valproate for three months.The frequency and duration of each seizure,the Montreal Cognitive Assessment Scale(MoCA),and the Quality of Life Rating Scale(QOLIE-31)were recorded before and after treatment.RESULTS There were 85 patients included(42 in the control group and 43 in the experimental group).After treatment,the seizure frequency in the experimental group was significantly reduced(P<0.05),and seizure duration was shortened(P<0.01).The total MoCA score in the experimental group significantly increased compared to before treatment(P<0.01),and the sub-item scores,except naming and abstract generalization ability,significantly increased(P<0.05),whereas the total MoCA score in the control group significantly decreased after treatment(P<0.05).The QOLIE-31 score of the experimental group increased significantly after treatment compared to before treatment(P<0.01).CONCLUSION Baijin pills have a good clinical effect on epilepsy with cognitive dysfunction.

Effect of dietary with Zhibai dihuang pills and gonadotropinreleasing-hormone-analogue on girls with precocious and rapidly progressive puberty

BACKGROUND At present,the clinical mechanisms underlying precocious puberty remain unclear,making effective intervention for children experiencing this condition and rapidly progressive puberty essential.AIM To explore the effects of Zhibai dihuang pills and gonadotropin-releasing hormone analogue(GnRHa)on growth and ovarian function in girls with precocious puberty.METHODS The clinical data of 84 adolescent girls with precocious puberty and rapidly progressive puberty from February 2017 to August 2023 were retrospectively analyzed.Girls were divided into a control group and an observation group,with 42 cases in each group.The control group received diet intervention combined with GnRHa treatment,while the observation group received diet intervention combined with Zhibai dihuang pills+GnRHa treatment.Outcomes such as clinical efficacy,growth indicators,ovarian function,and adverse reactions were compared between the two groups.RESULTS The observation group showed superior clinical efficacy compared to the control group(P<0.05).Prior to the intervention,no significant differences were found in growth or ovarian function between the groups(P>0.05).Post-intervention,the observation group exhibited significantly lower rates in growth,height,and bone age,along with reduced levels of progesterone,testosterone,estradiol,prolactin,luteinizing hormone,and follicle-stimulating hormone compared to the control group(P<0.05).The incidence of adverse reactions was similar across both groups(P>0.05).CONCLUSION Combining Zhibai dihuang pills with GnRHa and dietary intervention effectively improves growth,enhances ovarian function,and minimizes adverse reactions in adolescent girls with precocious and rapidly progressive puberty.

Application of Guizhi Fuling Pill in Gynecology

This paper reviews the research progress of Guizhi Fuling Pill in the clinical application of gynecology,such as the treatment of uterine leiomyoma,ovarian cyst,infertility and dysmenorrhea,in order to provide further research ideas for clinical researchers.

Determination of Plasma Concentration of Cinnamic Acid by High-Performance Liquid Chromatography and Its Pharmacokinetics in Rats after Oral Administration of Zi-Shen Pill

Aim To develop a simple and sensitive high-performance liquid chromatographicmethod for determination of plasma concentration of cinnamic acid and pharmacokinetic study in ratsafter a single oral dose of traditional Chinese medicinal preparation Zi-Shen pill. Method Plasmasamples were acidified with hydrochloric acid and extracted with ethyl acetate . Cinnamic acid wasdetermined by HPLC using a G_(18) column. A mobile phase ofmethanbl-acetonitrile-water-triethyl-amine (7:22:73 = 0.2, V/V), with the pH adjusted to 4.0 withphosphoric acid, and with a UV detector set at 340 nm. Results The standard curve was linear overthe range of 1.92- 192.0 μg·mL^(-1). The LLOQ was 1.92 μg·mL^(-1) . The RSDs of within-day andbetween-day precision were < 8%. The mean recovery was 82.0% . Conclusion After validation, themethpd has been used to investigate the pharmacokinetic profiles of the traditional Chinesemedicinal preparation Zi-Shen pill.

RP-HPLC Determination of Pinoresinol Diglucopyranoside in Qing′e Pill Extract

Aim To develop and determine pinoresinol diglucopyranoside in Qing'e Pill, atraditional Chinese compound preparation containing Eucommia ulmoides Oliv. as the principal drug,by a reverse-phase high-performance liquid chromatographic method (RP-HPLC) . Methods The extract ofQing'e Pill was refluxed with 75% ethanol, purified on an AB-8 macroporous adsorption resin columnand then injected into HPLC system. The HPLC assay was performed on an ODS analytical column with amixture of methanol-acetonitrile-water (24:3:78, V/V/V) as the mobile phase at a flow-rate of 1.0mL·min^(-1), and a UV detector set at 227 nm. Results Good linearity between peak area andconcentration was found in the range of 5.5 - 170 μg·mL^(-1) for pinoresinol diglucopyranoside ( r> 0.9998) . The average recovery was 99.3%. The intra-day assay RSD and the inter-day assay RSDwere 1.3% and 2.8%, respectively (n = 5). The content of pinoresinol diglucopyranoside in Qing'ePill was determined to be 0.446 +- 0.012 mg·g^(-1) (n = 10). Conclusion The RP-HPLC method wasproved to be sensitive, specific, accurate and precise for the determination of pinoresinoldiglucopyranoside in Qing' e Pill.

INVESTIGATION ON DRYING CHARACTERISTIC OF MEDICAL PILL USING MICROWAVE-CONVECTIVE DRYING

The combined microwave-convective drying is a novel drying method and has many advantages, such as swift drying, equable drying and sterilization. This paper studies the drying characteristic of medical pill in microwave-convective drying process. The influential parameters of pill drying characteristics including pill size, quality, air velocity, temperature and the input power of microwave, are experimentally investigated. Results show that the drying process is influenced by the intrinsic heat of microwave power, and the environment and the resistance of mass transfer. It has the advantage of the microwave-convective drying in saving energy, reducing labor intensity and sterilizing. The method with the high efficiency is suitable for pharmacy industry.

通脉开窍丸治疗血管性痴呆模型大鼠海马区神经元的铁死亡变化

背景:研究表明铁死亡与血管性痴呆存在密切联系,通脉开窍丸对于改善血管性痴呆患者的认知功能有一定疗效,但其作用机制不明确。目的:基于核因子E2相关因子2(nuclear factor erythroid-2 related factor 2,Nrf2)/血红素氧合酶1(heme oxygenase-1,HO-1)/谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)信号通路调控铁死亡探讨通脉开窍丸对血管性痴呆干预作用以及分子机制。方法:84只雄性SD大鼠,其中12只大鼠用作假手术组,其余大鼠用改良2-VO法制备成血管性痴呆的模型,成模后随机分为模型组、通脉开窍丸高、中、低剂量(27.6,13.8,6.9 g/kg)组、联合组[通脉开窍丸高剂量+ML385(20 mg/kg)]、盐酸多奈哌齐组(0.45 mg/kg),灌胃给药1次/d,联合组同时腹腔注射Nrf2抑制剂ML385,1次/d,连续4周。采用Morris水迷宫检测大鼠的学习记忆能力;苏木精-伊红染色观察各组大鼠海马组织中神经元病理学变化;比色法试剂盒检测大鼠血清中还原型谷胱甘肽、Fe^(2+)、丙二醛的浓度;普鲁士蓝染色法检测大鼠海马组织中铁沉积情况;透射电镜观察大鼠海马组织中神经元线粒体超微结构变化;蛋白免疫印迹法检测大鼠海马神经元Nrf2、HO-1、GPX4、XCT、铁蛋白重链1(ferritin heavy chain 1,FTH1)蛋白的表达。结果与结论:(1)与假手术相比,模型组大鼠逃避潜伏期时间明显延长(P<0.05),穿越平台次数明显减少(P<0.05);海马组织松散,神经元细胞核深染,染色质固缩甚至裂解;CA1区铁离子聚集;线粒体萎缩变小,线粒体嵴溶解消失,线粒体膜密度增厚;血清中Fe^(2+)、丙二醛水平上升,还原型谷胱甘肽水平下降(P<0.05);海马组织GPX4、HO-1、XCT、Nrf2、FTH1蛋白表达显著降低(P<0.05)。(2)与模型组相比,通脉开窍丸各剂量组和盐酸多奈哌齐组大鼠平均逃避潜伏期均明显缩短(P<0.05),穿越平台次数增加(P<0.05);海马神经元恢复明显,CA1区神经元铁离子聚集明显减少,线粒体结构和功能好转;血清Fe^(2+)、丙二醛水平显著降低(P<0.05),血清还原型谷胱甘肽浓度及海马组织中GPX4,HO-1,XCT,Nrf2,FTH1蛋白表达显著升高(P<0.05)。(3)与通脉开窍丸高剂量组相比,盐酸多奈哌齐组治疗效果差异无显著性意义(P>0.05),联合组大鼠水迷宫逃避潜伏期时间延长(P<0.05),穿越平台次数减少(P<0.05),大鼠CA1区神经元病理改变情况不明显,铁沉淀增加,血清中丙二醛、Fe^(2+)浓度增加(P<0.05),还原型谷胱甘肽浓度减少(P<0.05),海马组织神经元线粒体萎缩变小,且Nrf2、XCT、HO-1、GPX4、FTH1蛋白的表达减少(P<0.05)。在一定范围内,通脉开窍丸剂量越高效果越好,且高剂量治疗效果不亚于盐酸多奈哌齐。(4)结果说明,通脉开窍丸可以减轻大鼠海马组织神经元病理改变,改善血管性痴呆大鼠的认知功能,其作用机制可能与Nrf2/HO-1/GPX4信号通路的激活抑制铁死亡有关。

PillGrade 3D起球自动评级检测系统的探讨

本文介绍了起球自动评级检测系统的原理及评价过程。评定结果与主观评定方法相比,可以避免因试样的花型、颜色不同而引起的评级差异,使评定结果更加准确、客观。

西瓜嗜酸菌pilL菌毛基因的序列分析

在西瓜嗜酸菌标准菌株AAC00-1全基因组序列中选取了1个菌毛基因pilL序列,利用这个基因设计引物,选取国内外比较有代表性的7株西瓜嗜酸菌菌株进行PCR扩增,并对PCR产物进行了测序,从生物信息学的角度对这7个菌株pilL基因的序列特征、同源性、分子聚类、信号肽、疏水性、跨膜结构和二级蛋白结构进行了预测分析。结果表明:Aac5、pslbtw37、W1和pslbtw22 pilL基因之间的核苷酸氨基酸特征相近,同源性高,AAC00-1、AAC203-50和Fc376之间亦如此。预测发现7个编码蛋白表现为亲水性,都无跨膜结构,Aac5、pslbtw37和W1 pilL基因都存在一个类型为SPI的信号肽,其分泌蛋白所到达的亚细胞位点为s(即分泌到周质空间)。

Effect of Fufang Danshen Pill on Bone Marrow Stem Mobilization when Myocardial Scathe

Objective：To investigate the effect of Fufang Danshen pill on bone marrow stem mobilization during myocardial scathe. Methods：Rat models with expansionary myocardial disease were established by Pituitrin and Furazolidone. Experimental rats were divided into the contrast group, the myocardial scathe group （MS group）, the myocardial scathe and Fufang Dansben pill group （ MS ＋ FD group） and the myocardial scathe and fluvastatin group （ MS ＋ FT group）. The ratio of CD34^＋ cells was examined at the 1^st, 3^nl and 6^th weekend. Index of heart structure and function including LVESD, LVEDD. LYEF, LVEDP and dp/dtmax were evaluated at the 6^th weekend. The HW/BW index was calculated. Results：In the MS group, the index of HW/BW, LVESD, LVEDD and LVEDP were obviously increased （P 〈 0.01 ） and index of dp/ dtmax and LVEF were obviously decreased （P 〈 0.05 ）. The ratio of CD34^＋ cells was significantly improved at the 1^at weekend and then reduced slowly with no difference from that of the contrast group at the 6th weekend. Compared the MS ＋ FD group and the MS ＋ FT group with the MS group, the index of HW/BW, LYESD, LYEDD and LYEDP of were signifi cantly decreased （ P 〈 0.05 ） and index of dp/dtmax and LVEF were increased （P 〈 0.01 ）. The ratio of CD34^＋ cells was significantly higher at the 1^st, 3^nl and 6^th weekend, but had no statistic meaning at 3^nl and 6^th weekend （P 〉 0.05 ）. Conclusion：Pituitrin and Furazolidone can be used to establish rat models with expansionary myocardial disease. There has bone marrow stem mobilization during the early period of myocardial scathe. Fufang Danshen pill has effect on improving bone marrow stem mobilization, lightening the expansionary degree of heart and protecting the heart function. The effect of Fufang Danshen pill is as same as that of fluvastatin.

二仙丸加减方含药血清抑制巨噬细胞焦亡的物质基础和作用机制

背景:课题组前期研究发现二仙丸加减方可以缓解胶原诱导性关节炎大鼠炎症反应,但其作用机制尚需进一步验证。目的:分析二仙丸加减方入血成分,观察二仙丸加减方含药血清对J774A.1巨噬细胞焦亡的影响。方法:①二仙丸加减方入血成分分析:采用超高效液相色谱-高分辨质谱(UHPLC-HRMS)对二仙丸加减方及入血成分进行检测和鉴定。②二仙丸加减方含药血清对J774A.1巨噬细胞焦亡的影响:采用分子对接技术对二仙丸加减方入血成分倍半萜类化合物与NLRP3进行初步验证。将J774A.1巨噬细胞随机分为空白对照组、脂多糖+三磷酸腺苷组及脂多糖+三磷酸腺苷+二仙丸加减方含药血清低(2.5%)、中(5%)、高(10%)剂量组。根据试剂盒说明书检测各组细胞上清液中乳酸脱氢酶释放情况;ELISA检测各组细胞上清液中白细胞介素1β、白细胞介素18水平;Hoechst/PI染色法检测各组细胞膜受损情况;Western blot检测各组细胞中NLRP3、Caspase-1、GSDMD及GSDMD-N蛋白表达水平。结果与结论:①共鉴定出二仙丸加减方药效成分32个,入血成分21个,其中入血成分主要包括多种倍半萜类化合物;②分子对接结果显示3-O-Acetyl-13-deoxyphomenone、Incensol oxide、Atractylenolide Ⅲ、Rupestonic acid、3,7-Dihydroxy-9,11-eremophiladien-8-one与NLRP3之间结合活性较好;③与空白对照组相比,脂多糖+三磷酸腺苷组细胞上清中乳酸脱氢酶活性及白细胞介素1β、白细胞介素18水平均显著升高(P<0.001),Hoechst/PI染色可见PI阳性细胞数量显著增加。脂多糖+三磷酸腺苷+二仙丸加减方含药血清各组上述指标均显示不同程度降低;④与空白对照组相比,脂多糖+三磷酸腺苷组NLRP3、Caspase-1、GSDMD及GSDMD-N蛋白表达显著升高(P<0.05);与脂多糖+三磷酸腺苷组相比,脂多糖+三磷酸腺苷+二仙丸加减方含药血清各组细胞中NLRP3、Caspase-1、GSDMD及GSDMD-N蛋白表达均有不同程度降低(P<0.05),且具有一定的剂量依赖性。结果表明:二仙丸加减方含药血清可能通过调控NLRP3/Caspase-1/GSDMD通路抑制J774A.1巨噬细胞焦亡。

Effects of Fufang Biejia Ruangan Pills on hepatic fibrosis in vivo and in vitro

AIM:To explore the protective effect and the relevant mechanisms of Fufang Biejia Ruangan Pills(FFBJRGP)on hepatic fibrosis in vivo and in vitro.METHODS:Hepatic fibrosis was induced by carbon tetrachloride composite factors.Adult Wistar rats were randomly divided into four groups:normal control group;hepatic fibrosis model group;FFBJRGP-treated group at a daily dose of 0.55 g/kg;and colchicinetreated group at a daily dose of 0.1 g/kg.The effects of FFBJRGP on liver function,serum levels of hyaluronic acid(HA),typeⅣcollagen(CⅣ),typeⅢprocollagen(PCⅢ),laminin(LN),histopathology,and expression of transforming growth factor(TGF-β1)and Smad3 in hepatic fibrosis were evaluated in vivo.The effects of FFBJRGP on survival rate,hydroxyproline content and cell cycle distribution were further detected in vitro.RESULTS:Compared with the hepatic fibrosis model group,rats treated with FFBJRGP showed a reduction in hepatic collagen deposition and improvement in hepatic lesions.Compared with those of the model group,the activities of alanine aminotransferase(62.0±23.7 U/L)and aspartate aminotransferase(98.8±40.0 U/L)in the FFBJRGP-treated group were decreased(50.02±3.7 U/L and 57.2±30.0 U/L,respectively,P<0.01).Compared with those in the model group,the levels of PCⅢ(35.73±17.90 g/mL),HA(563.82±335.54 ng/mL),LN(89.57±7.59 ng/mL)and CⅣ(29.20±6.17ng/mL)were decreased to 30.18±9.41,456.18±410.83,85.46±7.51 and 28.02±9.45 ng/mL,respectively.Reverse-transcriptase polymerase chain reaction and Western blotting also revealed that expression of TGF-β1 and Smad3 were down-regulated in vivo.Cell proliferation was inhibited,the level of hydroxyproline was decreased compared with the control group(P<0.01),and the cell cycle was redistributed when exposed to FFBJRGP in vitro.CONCLUSION:FFBJRGP inhibits hepatic fibrosis in vivo and in vitro,which is probably associated with downregulation of fibrogenic signal transduction of the TGF-β-Smad pathway.

Study on prevention effect of Zishen Yutai pill combined with progesterone for threatened abortion in rats

Objective:To observe preventive and therapeutic effects and the mechanism of actions on Zishen Yutai pill combined with progesterone on threatened abortion in rats.Methods:After pregnancy,50 SPF female SD rats were selected and divided into control group,model group,progesterone group,Zishen Yutai pill group,and progesterone plus Zishen Yutai pill group(combination group),with 10 rats in each group.The rats of control group and abortion model group were lavaged with 2 mL/kg normal saline on pregnancy day 1 for continuous 10 days.Rats in Zishen Yutai pill group were given 1.575 g/kg/d of Zishen Yutai pill for intragastric administration for continuous 10 days.Rats in progesterone group were given intramuscular injection treatment of 0.1 mL/d progestin,continuous for 10 days.Rats in combination group were given injection therapy of aqueous solution of Zishen Yutai pill for continuous 10 days,and other treatments were the same as previous two groups.Abortion model were established then and live births,numbers of abortion and average rate of abortion were compared between the five groups.Peripheral blood was collected to detect the estradiol(E2) and progestational hormone(P),and obtain ratio of Th1/Th2 cytokines(IL-2,INF-γ,IL-4,IL-10).Results:Significant more live births of rats were found in the control group compared with other four groups(P<0.05).The numbers of live births of the rats in abortion model group were significantly less than that of progesterone group,Zishen Yutai pill group and joint group(P<0.05).The numbers of live births of rats in joint group were significant more than that of progesterone group and Zishen Yutai pill group.The serum E2 level of P of rats in the control group,progesterone group,Zishen Yutai pill group and joint group were significant higher than that of abortion model group(P<0.05).Serum levels of E2 and P of rats in the control group,progesterone group,Zishen Yutai pill group and joint group were not significant different(P>0.05) but these levels in the control group and Zishen Yutai pill group were significant lower than that of progesterone group and joint group(P<0.05).The maternal-fetal interface IL-4/IL-2,IL-10 / IL-2 of model group were balanced deviating to Th1,while the IL-4/IL-2,IL-10/IL-2 of progesterone group,Zishen Yutai pill group and joint group were balanced deviating to Th2,and joint group' deviation was better than progesterone group and Zishen Yutai pill group(P<0.05).Conclusions:Zishen Yutai pill combined with progesterone has a significant control effect for threatened abortion,which can obviously increase contents of maternal serums E2 and P,and regulate the Thl/Th2 balance with a remarkable effect.

Mechanism of action of Zhuyu Annao pill in mice with cerebral intrahemorrhage based on TLR4

Objective:To explore the protective effect and possible mechanism of action of Zhuyu Annao pill in mice with intracerebral hemorrhage(ICH).Methods:Sixty mice were divided into the control group,hemorrhage group,drug-treated group(after hemorrhage),TLR4-knockout hemorrhage group and TLR4-knockout hemorrhage + drug-treated group(after hemorrhage) with 12 in each group.Model of autologous ICH was established in all groups.After drilling and 12 h of fasting,models in the control group hemorrhage group and TLR4-knockout hemorrhage group were all drenched with 10 mL/kg distilled water by intragastric administration.Models in the drug-treated group and TLR4-knockout hemorrhage + drugtreated group were drenched with 6.25 g/kg of Zhuyu Annao pill.All groups were treated for 7 d.Longa scoring method was used to measure the neurological defect scores and determine the brain water contents of all groups;ELISA was employed to detect the inflammatory factor interleukin(IL)-6,tumor necrosis factor- α(TNF- α) and IL-1β in brain tissues;and Western blot was applied to test the expression quantities of apoptotic protein Bax and anti-apoptotic protein Bcl-2 in brain tissues.Results:At day 3 and7,compared with the hemorrhage group,the neurological defect scores of the drug-treated group,TLR4-knockout hemorrhage group and TLR4-knockout hemorrhage + drug-treated group decreased significantly(P<0.05) Compared with the hemorrhage group,the brain water contents of the drug-treated group,TLR4-knockout hemorrhage group and TLR4-knockout hemorrhage + drug-treated group reduced significantly(P<0.05) Compared with the hemorrhage group,the inflammatory factor IL-6,TNF-α and IL-1β of the drug-treated group,TLR4-knockout hemorrhage group and TLR4-knockout hemorrhage + drug-treated group decreased significantly(P<0.05).Compared with the hemorrhage group,the expression of apoptotic protein Bax of the drug-treated group,TLR4-knockout hemorrhage group and TLR4-knockout hemorrhage+ drug-treated group decreased significantly and the expression of anti-apoptotic protein Bcl-2 increased significantly(P<0.05).Conclusions:Zhuyu Annao pill can alleviateencephaledema for mice with ICH and reduce inflammatory responsesandnerve cell apoptosis.TLR4 can mediate inflammatory injury induced by ICH.Thus,Zhuyu Annao pill can play a protective role for brains by decreasing the expression of TLR4.