Erxian decoction potentially prevents postmenopausal osteoporosis by modulating miR-335: a study based on bioinformatics analysis and preliminary clinical case validation

Background:miRNAs are closely related to bone metabolism.Studies have shown that Erxian decoction can improve bone metabolism,possibly achieving this regulatory effect through miRNA targets.Netinfer was used to predict the miRNA targets of Erxian decoction for the treatment of postmenopausal osteoporosis,and the results were validated by clinical trials.Methods:In this study,we identified possible targets of Erxian decoction in osteoporosis by means of network pharmacological analysis and bioinformatic prediction.Fifteen cases of postmenopausal osteoporosis with kidney Yin and Yang deficiency(In traditional Chinese medicine,kidney Yin nourishes and moistens the tissues of the internal organs of the body,while kidney Yang promotes and warms the tissues of the internal organs of the body.)were treated with Erxian decoction for four weeks,and serum bone metabolism indices(P1NP,osteocalcin,andβ-CTX)and miRNA-335-5p expression were measured before and after treatment.Results:The constructed miRNA postmenopausal osteoporosis related gene network of the effective compound of the Erxian decoction has 296 points and 981 edges.The 39 postmenopausal osteoporosis related genes regulated by miRNA-335-5p were enriched in ossification,while the signaling pathways were enriched in rheumatoid arthritis,the Toll signaling pathway,the HIF-1 signaling pathway,and the MAPK signaling pathway.After taking Erxian decoction,the expression of the serum bone formation index(P1NP,osteocalcin)and miRNA-335-5p gene expression levels increased significantly.The alterations in P1NP and osteocalcin were correlated with the changes in miRNA-335-5p.Conclusion:Circulating miRNA-335-5p may serve as an important target of Erxian decoction in the treatment of postmenopausal women.The effect of Erxian decoction on bone formation is significant,but the underlying mechanism requires further investigation.

益气活血解毒汤对动脉硬化患者血清MMP-9、CRP及血脂的影响

目的观察益气活血解毒汤对动脉粥样硬化(AS)患者治疗前后血清基质金属蛋白酶-9(MMP-9)、C反应蛋白(CRP)及血脂的变化,探讨其对AS稳定性的影响及相关机制。方法患者随机分为两组,对照组20例,采用常规西药治疗,治疗组20例,在对照组治疗的基础上加服益气活血解毒汤,疗程均为3周。治疗前后分别测定两组MMP-9、CRP、甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)以及高密度脂蛋白胆固醇(HDL-C)。结果对照组治疗后MMP-9、CRP、TG、TC、LDL-C及HDL-C变化不明显,治疗组治疗后HDL-C增高,MMP-9、TC、LDL-C、CRP明显下降。治疗组TC、LDL-C、CRP、MMP-9和HDL-C的改善优于对照组。结论益气活血解毒汤可稳定AS斑块,其机制可能为调脂、抗炎、抑制基质金属蛋白酶。

Huangqi Decoction,a compound Chinese herbal medicine,inhibits the proliferation and activation of hepatic stellate cells by regulating the long noncoding RNA-C18orf26-1/microRNA-663a/transforming growth factor-βaxis

Objective Huangqi Decoction(HQD),a classical traditional Chinese medicine formula,has been used as a valid treatment for alleviating liver fibrosis;however,the underlying molecular mechanism is still unknown.Although our previous studies showed that microRNA-663a(miR-663a)suppresses the proliferation and activation of hepatic stellate cells(HSCs)and the transforming growth factor-β/small mothers against decapentaplegic(TGF-β/Smad)pathway,whether long noncoding RNAs(lncRNAs)are involved in HSC activation via the miR-663a/TGF-β/Smad signaling pathway has not yet reported.The present study aimed to investigate the roles of lncRNA lnc-C18orf26-1 in the activation of HSCs and the mechanism by which HQD inhibits hepatic fibrosis.Methods The expression levels of lnc-C18orf26-1,miR-663a and related genes were measured by quantitative reverse transcription-polymerase chain reaction.HSCs were transfected with the miR-663a mimic or inhibitor and lnc-C18orf26-1 small interfering RNAs.The water-soluble tetrazolium salt-1 assay was used to assess the proliferation rate of HSCs.Changes in lncRNA expression were evaluated in miR-663a-overexpressing HSCs by using microarray to identify miR-663a-regulated lncRNAs.RNA hybrid was used to predict the potential miR-663a binding sites on lncRNAs.Luciferase reporter assays further confirmed the interaction between miR-663a and the lncRNA.The expression levels of collagen α-2(I)chain(COL1A2),α-smooth muscle actin(α-SMA)and TGF-β/Smad signaling pathway-related proteins were determined using Western blotting.Results Lnc-C18orf26-1 was upregulated in TGF-β1-activated HSCs and competitively bound to miR-663a.Knockdown of lnc-C18orf26-1 inhibited HSC proliferation and activation,downregulated TGF-β1-stimulatedα-SMA and COL1A2 expression,and inhibited the TGF-β1/Smad signaling pathway.HQD suppressed the proliferation and activation of HSCs.HQD increased miR-663a expression and decreased lnc-C18orf26-1 expression in HSCs.Further studies showed that HQD inhibited the expression of COL1A2,α-SMA,TGF-β1,TGF-βtype I receptor(TGF-βRI)and phosphorylated Smad2(p-Smad2)in HSCs,and these effects were reversed by miR-663a inhibitor treatment.Conclusion Our study identified lnc-C18orf26-1 and miR-663a as promising therapeutic targets for hepatic fibrosis.HQD inhibits HSC proliferation and activation at least partially by regulating the lnc-C18orf26-1/miR-663a/TGF-β1/TGF-βRI/p-Smad2 axis.

益气活血解毒汤对心肌缺血大鼠TNF-α、IL-6及心肌组织的影响

目的观察益气活血解毒汤对实验性心肌缺血大鼠肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)及心肌组织的影响,探讨其抗心肌缺血的作用机理。方法将70只Wistar大鼠随机分为空白对照组,模型组,益气活血解毒汤低、中、高剂量组及复方丹参滴丸组,硝酸异山梨酯组,每组10只。采用腹腔注射异丙肾上腺素(ISO)制造心肌缺血模型,记录各组大鼠心电图,造模后30 min处死动物,测定TNF-α、IL-6,取心尖部全层心肌观察心肌损伤情况。结果益气活血解毒汤可以降低心肌缺血大鼠J点总偏移值;降低血清TNF-α、IL-6水平,减轻心肌缺血导致的病理损伤,以高剂量效果最为明显。结论益气活血解毒汤对ISO造成的心肌缺血大鼠具有保护作用,其作用机制可能与其抑制炎症反应,降低TNF-α、IL-6的水平有关。

益气活血解毒汤对实验性动脉粥样硬化家兔血脂、NO、ET-1、MDA及SOD的影响

目的观察益气活血解毒汤对实验性动脉粥样硬化(AS)家兔血脂、一氧化氮(NO)、内皮素(ET-1)、脂质过氧化产物(MDA)及超氧化物歧化酶(SOD)的影响。方法24只雄性日本大耳白兔随机分为空白对照组、模型对照组、益气活血解毒汤组、辛伐他汀组,高脂饲料喂饲复制AS模型;10周后测定各组家兔上述指标。结果与模型对照组相比,益气活血解毒汤组TG、TC、LDL、MDA、ET-1含量明显下降,NO、HDL含量明显升高,SOD活性升高。结论益气活血解毒汤有明显的抗家兔AS形成的作用,可能与其降低血脂、改善内皮细胞功能、减轻过氧化损伤等因素有关。

Chaihushugan decoction exerts antiepileptic effects by increasing hippocampal glutamate metabolism in pentylenetetrazole-kindled rats

OBJECTIVE: To investigate the antiepileptic effects of Chaihushugan decoction(CHSGD) in rats with pentylenetetrazole(PTZ)-induced seizures and to discuss the impact of CHSGD on glutamate metabolism, a hypothesized underlying mechanism of seizure reduction.METHODS: Fifty Wistar rats were divided randomly into either control(n = 10) or experimental(n = 40)groups. Rats in the control group were administered physiological saline intraperitoneally. A subconvulsive dose of PTZ(35 mg/kg) was administered intraperitoneally to rats in the experimental group to induce seizures. The fully PTZ-kindled rats were then randomly divided into five subgroups(n = 8 each) based on the following treatment categories: physiological saline, VPA(200 mg/kg), CHSGD(2.5 g/kg), CHSGD(5 g/kg), or CHSGD(10 g/kg),administered orally once per day, respectively. On day 28 following initiation of drug treatment, seizures were monitored. The rats were then sacrificed, and hippocampal dissections were performed for subsequent studies.RESULTS: CHSGD significantly prolonged the latency of myoclonic, clonic, and tonic seizures, while decreasing overall seizure rates in the kindled rats.The measured concentrations of 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxy-d-glucose(2-NBDG) and glutamate were significantly lower in the hippocampi of kindled rats in groups treated with CHSGD compared with those treated with PTZ alone. In addition, CHSGD was found to up-regulate both the expression of glutamate transporter-1(GLT-1) protein and the activity of glutamine synthetase(GS) in the hippocampi of kindled rats.CONCLUSION: These results suggest that CHSGD has antiepileptic effects on PTZ-induced seizures.The results further suggest an increase in glutamate metabolism at the synaptic cleft is a putative underlying mechanism of seizure reduction.

Fuzheng Kang'ai decoction(扶正抗癌方) inhibits cell proliferation,migration and invasion by modulating mir-21-5p/human phosphatase and tensin homology deleted on chromosome ten in lung cancer cells

OBJECTIVE:To elucidate the potential molecular mechanism by which Fuzheng Kang’ai decoction(扶正抗癌方,FZKA)inhibits proliferation,migration,and invasion of lung cancer cells.METHODS:Varying FZKA concentrations were used to manage lung cancer cells(A549 and PC9).We employed:cell counting kit-8(CCK-8)and plate clone formation assays to examine the cell viability;flow cytometry(FCM)to analyze the cycle arrest;transwell and woundhealing assays to assess the cell invasion and migration,respectively.Further,a quantitative real-time polymerase chain reaction(q RT-PCR)assay was adopted to evaluate the miR-21-5p expression.For protein expression analysis,we employed the Western blot technique.Recombinant miR-21-5p overexpression adenovirus vector harboring GFP was constructed and transfected into A549 and PC9,after which we explored the effect of FZKA on miR-21-5p overexpression.RESULTS:Notably,treatment with FZKA inhibited viability,clone-formation ability,invasion,and migration of lung cancer cells.Mechanistically,FZKA markedly suppressed miR-21-5p expression but elevated the human phosphatase and tensin homology deleted on chromosome ten(PTEN)protein level in both A549 and PC9 cells.Over-expression of miR-21-5p lowered PTEN protein expression.Besides,overexpressed miR-21-5p levels with adenovirus antagonized FZKA-upregulated PTEN protein expression.CONCLUSION:The present study demonstrates how FZKA modulates cell biological behaviors,for instance,it impedes the proliferation by upregulating PTEN expression with miR-21-5p as the target.These findings unveil the potential novel molecular mechanisms from the micro RNA aspect by which FZKA suppresses the growth of human lung cancer cells.

益气活血解毒汤对实验性动脉粥样硬化家兔C反应蛋白及血脂水平的影响

目的利用高脂喂饲诱发动脉粥样硬化模型,探讨中药复方益气活血解毒汤对兔动脉粥样硬化的干预作用及其机制。方法24只日本雄性大耳白兔随机分为4组,每组6只,正常对照组、模型对照组、益气活血解毒汤组、辛伐他汀组。其中益气活血解毒汤组及辛伐他汀组边制模边给药,而前两组以等量蒸馏水灌胃。10周后,测定各组空腹血清三酰甘油(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)以及C反应蛋白(CRP)含量。结果模型对照组CRP、TG、TC及LDL-C较正常对照组有不同程度升高,HDL-C下降(P<0.05或P<0.01)。益气活血解毒汤组及辛伐他汀组CRP、TG、TC及LDL-C均明显低于模型对照组,HDL-C升高明显(P<0.01),而与正常对照组比较无统计学意义(P>0.05)。益气活血解毒汤组HDL-C与辛伐他汀组比较有所升高,差异有统计学意义(P<0.05)。结论益气活血解毒汤对动脉粥样硬化有明显的预防和治疗作用。降低CRP和调节血脂可能是其作用机制之一。

益气活血解毒汤对实验性动脉粥样硬化家兔血脂、前列环素及血栓素A_2的影响

目的通过高脂饲料喂饲诱发动脉粥样硬化家兔模型,探讨中药复方益气活血解毒汤对家兔动脉粥样硬化的干预作用及其作用机制。方法24只日本雄性大耳白兔随机分为4组,分别为空白对照组、模型对照组、益气活血解毒汤组及辛伐他汀组。10周后测定各组血清总胆固醇(TC)、三酰甘油(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、血栓素B2(TXB2)、6-酮-前列腺素(6-keto-PGF1α)的含量。结果模型对照组TC,TG,LDL-C,TXB2较空白对照组有不同程度升高(P<0.05或P<0.01),6-keto-PGF1α及HDL-C显著下降(P<0.05)。益气活血解毒汤组及辛伐他汀组TC,TG,LDL-C及TXB2均低于模型对照组,6-keto-PGF1α及HDL-C升高明显(P<0.01),益气活血解毒汤组HDL-C与辛伐他汀组比较有所升高(P<0.05)。结论益气活血解毒汤对家兔动脉粥样硬化有明显的预防和治疗作用,调节血脂、维持PGI2/TXA2的平衡可能是其作用机制之一。

益气活血解毒方抑制家兔经皮血管腔内成形术后再狭窄的实验研究

目的:探讨益气活血解毒方对经皮血管腔内成形术(PTA)后再狭窄的防治作用。方法:采用电刺激兔颈总动脉加高脂饲料8周后,体表多普勒超声评价颈动脉粥样斑块狭窄,再行颈动脉PTA,形成再狭窄。造模后动物随机分为模型组、西药组(普伐他汀和巴米尔)、中药组(益气活血解毒方),另设正常组和假手术组作为对照,含药饲料治疗4周。分别在不同时间点检测血脂、血清C-反应蛋白(CRP)、转化生长因子(TGF-β)。药后4周颈总动脉取材,常规HE、弹力+VG和Masson染色,平滑肌肌动蛋白(α-SM-actin)和巨噬细胞RAM11免疫组化分析,所有病理切片经显微图像分析系统分析。结果:益气活血解毒方突出表现为对增生内膜中胶原的抑制作用,减少狭窄局部胶原面积比达50%;能减少血清中TGF-β的含量,降低血脂;而对血清CRP、增生内膜中巨噬细胞RAM11的表达无明显抑制作用。结论:益气活血解毒方通过减少增生内膜中胶原的堆积、降低血脂和TGF-β的含量,防治PTA后再狭窄。

益气活血解毒汤治疗冠心病稳定型心绞痛疗效观察

目的观察益气活血解毒汤配合常规西药治疗稳定型心绞痛临床疗效。方法将40例冠心病稳定型心绞痛患者按入选顺序分为研究组与对照组各20例,研究组在常规西医治疗基础上加服益气活血解毒汤治疗,对照组仅予常规西医治疗;比较两组心绞痛发作的次数、持续时间、疼痛程度及中医证候、静息心电图等的变化。结果治疗后两组心绞痛再发次数、持续时间、中医证候、心电图等较治疗前均有明显改善,而研究组改善优于对照组。结论益气活血解毒汤治疗冠心病稳定型心绞痛疗效满意。

益气活血解毒汤对实验性动脉粥样硬化家兔CRP水平及主动脉斑块的影响

目的观察中药复方益气活血解毒汤对实验性动脉粥样硬化(AS)家兔C反应蛋白(CRP)及主动脉斑块的干预作用及其机制。方法24只日本雄性大耳白兔随机分为正常对照组、模型对照组、益气活血解毒汤组、辛伐他汀组。采用高脂饲料喂饲建立兔AS模型,其中益气活血解毒汤组及辛伐他汀组造模同时灌胃相应药液,而前两组以等量蒸馏水灌胃。10周后测定各组家兔斑块面积及血清CRP的含量。结果模型对照组CRP较正常对照组升高,益气活血解毒汤组及辛伐他汀组CRP明显低于模型组,与正常对照组相近。模型对照组斑块面积较空白对照组明显升高,益气活血解毒汤组及辛伐他汀组斑块面积均低于模型对照组,益气活血解毒汤组斑块面积与辛伐他汀组比较有所降低。结论益气活血解毒汤对AS有明显的预防和治疗作用,其可减少斑块的面积,降低血清CRP含量,从而起到稳定AS斑块的作用。

益气活血解毒汤对实验性动脉粥样硬化家兔炎症细胞因子和血脂的影响

目的:观察中药复方益气活血解毒汤对实验性动脉粥样硬化(AS)家兔血脂及炎症细胞因子水平的影响,探讨该方防治AS的机制。方法:将24只日本雄性大耳白兔随机分为正常对照组、模型组、益气活血解毒汤组和辛伐他汀组,每组6只。采用高脂饲料喂饲建立兔AS模型,其中益气活血解毒汤组及辛伐他汀组在制模的同时分别给药,而正常对照组和模型组以等量蒸馏水灌胃。10周后测定各组空腹血清甘油三酯(TG)、总胆固醇(TC)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、C-反应蛋白(CRP)、白细胞介素-6(IL-6)及肿瘤坏死因子-α(TNF-α)含量。结果:模型组TG、TC、LDL-C、CRP、IL-6及TNF-α均较正常对照组有不同程度的升高,HDL-C明显下降,差异均有显著性(P均<0.01)。益气活血解毒汤组及辛伐他汀组TG、TC、LDL-C、CRP、IL-6及TNF-α均明显低于模型组,HDL-C则明显升高,差异均有显著性(P均<0.01),而与正常对照组比较差异无显著性(P均>0.05)。益气活血解毒汤组HDL-C较辛伐他汀组有所升高,差异有显著性(P<0.05)。结论:益气活血解毒汤对AS有明显的预防和治疗作用;调节血脂和降低炎症细胞因子水平可能是其作用机制之一。

益气活血解毒汤治疗老年稳定型心绞痛气虚血瘀证临床观察

目的观察益气活血解毒汤对老年冠心病稳定型心绞痛气虚血瘀证患者中医证候及生活质量的影响。方法将69例患者随机分为治疗组与对照组,均予西医常规治疗,治疗组加服益气活血解毒汤;比较两组中医证候积分以及总有效率,并采用西雅图心绞痛问卷量表对患者躯体活动受限程度、心绞痛稳定状态及发作情况、治疗满意程度和疾病认识程度进行测评。结果治疗组中医证候、总有效率以及生活质量的改善均优于对照组。结论益气活血解毒汤配合西医常规治疗对老年冠心病稳定型心绞痛的疗效优于单纯西医常规治疗。

益气活血解毒汤治疗慢性阻塞性肺疾病急性发作期疗效观察

目的观察中药益气活血解毒汤治疗慢性阻塞性肺疾病急性发作期的临床疗效。方法将78例慢性阻塞性肺疾病急性发作期患者随机分为治疗组43例与对照组35例,治疗组在西医常规治疗的基础上给予益气活血解毒汤剂,对照组予以西医常规治疗;比较两组临床疗效及血气分析、血常规、C超敏反应蛋白、降钙素原、肺功能等指标的变化。结果治疗组临床疗效及多项指标改善均优于对照组。结论益气活血解毒汤联合西药对慢性阻塞性肺疾病急性加重期有良好的治疗作用。

益气活血解毒汤治疗气虚血瘀型心绞痛的临床观察

目的观察益气活血解毒汤治疗气虚血瘀型心绞痛的临床价值。方法选取我院在2012年3月-2014年3月收治的患有气虚血瘀型心绞痛的140例患者,随机分成治疗组和对照组,对照组给予常规治疗,治疗组在此基础上给予益气活血解毒汤的治疗。结果治疗组的治疗总有效率为98.5%,对照组的治疗总有效率为74.2%,差异对比具有统计学意义（P〈0.05）。结论采用益气活血解毒汤治疗气虚血瘀型心绞痛,效果较好。

益气活血解毒汤联合西药治疗气虚血瘀型蛇串疮临床观察

目的:通过观察西医常规疗法联合益气活血解毒汤治疗气虚血瘀型蛇串疮的有效性及安全性,为益气活血解毒汤治疗气虚血瘀型蛇串疮提供一定的依据。方法:将符合纳入标准的64例气虚血瘀型蛇串疮患者,随机分为治疗组和对照组各32例。对照组给予西医常规疗法基础治疗,治疗组在对照组基础上联合益气活血解毒汤治疗,两组均治疗10 d后观察临床疗效。结果:治疗组总有效率优于对照组(P<0.05),且治疗组在改善临床症状方面优于对照组(P<0.05)。结论:西医常规疗法联合益气活血解毒汤治疗气虚血瘀型蛇串疮安全有效,值得临床推广。