Effect of Shuanghuanglian Oral Solution on Liver Function in a Mouse Model of Non-alcoholic Steatohepatitis

[Objectives]To observe the effect of Shuanghuanglian oral solution on liver function in BABL/cJ mice in non-alcoholic steatohepatitis(NASH)model.[Methods]The BABL/cJ mice were randomly divided into three groups:a control group,a model group,and an experimental group.The experimental group was administered with 10%Shuanghuanglian oral solution at a dose of 0.1 mL/(10 g·d),while the control group and experimental group received an equivalent dosage of normal saline.All three groups were treated for a period of 28 d.The liver function of the mice in each group was examined after the treatment.[Results]The body mass,liver index,triacylglycerol(TG),total cholesterol(TC),aspartate aminotransferase(AST),and alanine aminotransferase(ALT)levels were all significantly reduced compared to the model group(P<0.05).[Conclusions]Shuanghuanglian oral solution has a beneficial effect on liver function in BABL/cJ mice.

Rapid and Sensitive Assay of the Main Components in Shuanghuanglian Preparations by UPLC-TOF-MS

1 Introduction As already known, complex prescription has been the outstanding feature of traditional Chinese medicine（TCM）. This fact therefore brings complicated formula with multicomponents from multimaterals, which produces some difficulties to detect the components and to control the quality. Shuanghuanglian preparations（SHLs）, including oral liquid, injection powder, capsule, and tablet, belong to a family of refined traditional Chinese patent medicine and a combined herbal remedy comprised of three herbs：

Outlining the keyword co-occurrence trends in Shuanghuanglian injection research: A bibliometric study using CiteSpace Ⅲ

Objective:To explore the evolvement and new trends in the use of Shuanghuanglian injection(SHLI).Methods:China National Knowledge Infrastructure(CNKI),PubMed,and Embase were extensively searched using the search terms“Shuanghuanglian injection”and“Shuanghuanglian fenzhen”to retrieve articles relevant to SHLI(1992e2020).Retrieved articles were further investigated by two authors to exclude those unrelated to SHLI.The bibliographical references of the included articles were exported as raw data and then treated using the CiteSpace tool to visualize the mapping of the SHLI research domain.Essential clusters and highly frequent keywords were quantified for further analysis.The clusters were automatically labeled by the algorithm of tf*idf for objective analysis.Basic bibliometric features,including article types and yearly trend in article numbers were also determined and discussed.Results:The modules of the keywords of interest presented clear boundaries with a high modularity score(Q=0.73).High-confidence clusters were identified,including bioactivity fingerprint(S=0.99),equal pupils(S=0.91),drug preparation department(S=0.87),difficulty in respiration(S=0.85),peristalsis(0.88),and Danshen powder injection(S=0.94).The characteristic keywords in terms of frequency and burstiness were Shuanghuanglian powder for injection(F=235,B=5.22),SHLI(F=112,B=11.39),and adverse drug reactions(ADRs;F=104,B=7.35).Conclusion:In the field of SHLI study,there are five major topic categories:bioactivity fingerprint;ADR mechanism and cause detection;proper preparation;clinical evidence accumulation;and efficacy in diseases with no effective treatment and combination usage.The trend for using modern methodologies from a science-based perspective to study SHLI will continue to exist.The causes of multi-factorial ADRs may be an important topic for future studies.

Inhibitory effects of Shuanghuanglian injection on nuclear factor-kappa B expression in mice with viral encephalitis in a time-and dose-dependent manner

Previous studies have confirmed that the anti-virus effects of Shuanghuanglian injection may be associated with nuclear factor-kappa B activity. This study observed nuclear factor-kappa B expression in mice with viral encephalitis, and showed significant decreases in nuclear factor-kappa B protein and mRNA levels following Shuanghuanglian injection. The inhibitory effect was more significant with prolonged intervention duration and increased treatment dose. These findings verify that Shuanghuanglian injection plays a therapeutic role in viral encephalitis by reducing expression of nuclear factor-kappa B in a time- and dose-dependent manner.

Shuanghuanglian injection downregulates nuclear factor-kappa B expression in mice with viral encephalitis

A mouse model of viral encephalitis was induced by intracranial injection of a Coxsackie virus B3 suspension. Quantitative real-time reverse transcription-PCR and western blot assay were applied to detect mRNA and protein expression of intelectin-2 and nuclear factor-kappa B in the viral encephalitis and control groups. Nuclear factor-kappa B and intelectin-2 mRNA and protein expression were significantly increased in mice with viral encephalitis. After intraperitoneal injection of Shuanghuanglian at a dose of 1.5 mg/kg for 5 successive days, intelectin-2 and nuclear factor-kappa B protein and mRNA expression were significantly decreased. To elucidate the relationship between intelectin-2 and nuclear factor-kappa B, mice with viral encephalitis were administered an intracerebral injection of 107 pfu recombinant lentivirus expressing intelectin shRNA. Both protein and mRNA levels of intelectin and nuclear factor-kappa B in brain tissue of mice were significantly decreased. Experimental findings suggest that Shuanghuanglian injection may downregulate nuclear factor-kappa B production via suppression of intelectin production, thus inhibiting inflammation associated with viral encephalitis.

Effect of Shuanghuanglian Propolis Oral Liquid on Immune Function of Broilers

The effects on immune function of broilers was studied by adding different doses of Shuanghuanglian propolis oral liquid in drinking water, to provide a theoretical basis for application and promotion of Shuanghuanglian propolis oral liquid. Ninety one-day-old broilers were randomly divided into three groups ： group I was designed as control; groups Ⅱ and Ⅲ were added with 0.5 and 1.0 mL/L Shuanghuanglian propolis oral liquid in drinking water, respectively. Compared with group I, all tested indicators in groups Ⅱ and Ⅲ were increased at varying degrees as follows ： bursa of fabricius index increased by 5.74% （P 〈 0.05 ） and 7.66% （P 〈 0.05 ） ; thymus index increased by 1.47% （P 〉 0.05 ） and 2.06% （P 〈 0.05 ） ; spleen index increased by 5.88% （P 〉 0.05 ） and 7.84% （ P 〈 0.05 ） ; Newcastle disease virus （NDV） antibody titer increased by 11.99% （P 〈 0.05 ） and 16.94% （P 〈 0.05 ） ; IL-2 content increased by 2.71% （P 〉 0.05） and 4.52% （P 〈 0.05 ） ; IL -6 content increased by 4.04% （P 〉 0.05 ） and 7.26% （ P 〈 0.05 ）, respectively. The results showed that adding 0.5 and 1.0 mL/L Shuanghuangllan propolis oral liquid in drinking water could improve the immune function of broiler and immune effect of ND vaccine.

Effects of Shuanghuanglian Propolis Oral Liquid on Growth Performance and Intestinal Microflora of Broilers

[ Objective] The paper was to investigate the effects of adding different levels of Shuanghuanglian propelis oral liquid in drinking water on growth performance and intestinal microflora of broilers. [ Method] Ninety one-day-old broilers were randomly divided into three groups （group I, group 1I and group III）： group I was the control group, and groups II and III were added with 0.5 and 1.0 mL/L Shuanghuanglian prepolis oral liquid in drinking water, respec- tively. [ Result] Compared with broilers in group I, the average daily gains of broilers in group II and group III increased significantly （ P 〈 0.05 ） ; the feed gain ratio of broilers in group III was significandy lower than that in group I （ P 〈 0.05 ） ; the number of intestinal LactobaciUus spp. in group II and group III in- creased significantly （ P 〈 0.05 ） ; the number of Salmonella spp. and Escherichia coli in group lII decreased significantly （ P 〈 0.05 ） ; the number of Bgrdobacterium spp. increased significantly （ P 〈 0.05 ）. [ Conclusion] Adding 0.5 and 1.0 mL/L Shuanghuanglian propolis oral liquid in drinking water could improve growth performance and intestinal micreflora of broilers, and the dose of 1.0 mL/L is recommended in clinical application.

Study on the Effect of Shuanghuanglian Powder Needle Combined with Cefoperazone and Sulbactam Sodium on Pseudomonas aeruginosa in Vitro

Objective: To explore the antibacterial activity of combined use of Shuanghuanglian and cefoperazone sulbactam sodium on resistant strains of Pseudomonas aeruginosa. Methods: The Pseudomonas aeruginosa strains which were sensitive and resistant to cefoperazone sulbactam sodium were selected to prepare different test bacterial solutions respectively;The experimental liquid of Shuanghuanglian and Cefoperazone Sulbactam Sodium were prepared separately and set as different test groups and control groups;The Drug Sensitivity Tests of Shuanghuanglian and cefoperazone sulbactam sodium at different concentration gradients which were used alone or used in combination were carried out for different strains with sensitivity and resistance, And use standard entry as a reference control. Result: The results of drug sensitivity test of Shuanghuanglian combined with Cefoperazone-Sulbactam sodium against the resistant strains of Pseudomonas aeruginosa were compared with the results of drug sensitivity test of the two separately used, and the difference was statistically significant (P 〈 0.05) [The drug sensitivity test results of Shuanghuanglian and cefoperazone sulbactam sodium to Pseudomonas aeruginosa resistant strains were statistically significant compared with the drug sensitivity test results of Shuanghuanglian and Cefoperazone Sulbactam Sodium used separately (P 〈 0.05)];There was a dependence between strains and concentration in the effect of the combination of the two drugs. Conclusion: The combination of Shuanghuanglian and cefoperazone sulbactam sodium has synergistic antibacterial or bactericidal effect on Pseudomonas aeruginosa resistant strains. .

Component Analysis of Shuanghuanglian Freeze-dried Powder and Its Effects on Human Hepatocyte Function when Combined with Cefradine

[Objectives]To detect the contents of components in Shuanghuanglian freeze-dried powder and to explore the effects of Shuanghuanglian freeze-dried powder and its components on human hepatocytes(HL-7702)alone or in combination with cefradine.[Methods]High performance liquid chromatography(HPLC)was used to detect the contents of baicalin,wogonin,chlorogenic acid and forsythin,the main components of Shuanghuanglian freeze-dried powder.HL-7702 cells were cultured with Shuanghuanglian freeze-dried powder and the main components of Shuanghuanglian freeze-dried powder alone or in combination with cefradine.Enzyme linked immunosorbent assay(ELISA)was used to detect the contents of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in the cell supernatant after culture,and HPLC was used to detect the expression level of adenosine diphosphate(ADP)and adenosine triphosphate(ATP);agarose gel electrophoresis was used to detect the expression of cyclooxygenase 2(COX-2)and heme oxygenase-1(HO-1)in HL-7702 cells.[Results]In Shuanghuanglian freeze-dried powder for injection,the content of baicalin was the highest,and the content of wogonin was the lowest.Compared with the control group,the expressions of AST and ALT in human hepatocytes(HL-7702)in high-dose baicalin group,forsythin group and wogonin group decreased(P<0.05,P<0.01),while the expression of ALT in chlorogenic acid+cefradine group(0.046 mg/mL)and forsythin+cefradine group(0.046 mg/mL)increased(P<0.05,P<0.01),and the expression of AST had no significant difference(P>0.05);the results in the low-dose group were similar to those in the high-dose group.Compared with the control group,ATP expression in chlorogenic acid group,chlorogenic acid+cefradine group(0.046 mg/mL)and forsythin+cefradine group(0.046 mg/mL)in the high-dose group decreased(P<0.05,P<0.01),and ADP expression was not significantly different(P>0.05);in the low-dose group,the expression of ATP and ADP increased in baicalin group(P<0.05),but decreased in wogonin group,baicalin+cefradine group(0.046 mg/mL)and wogonin group+cefradine group(0.046 mg/mL)(P<0.05,P<0.01).Compared with the control group,the expressions of COX-2 and HO-1 in HL-7702 cells in the cefradine group showed no significant difference(P>0.05).The expression of HO-1 and COX-2 in the different dose groups of Shuanghuanglian and the group combined with cefradine increased,and the difference was significant(P<0.05,P<0.01).[Conclusions]The components of Shuanghuanglian freeze-dried powder for injection had effects on hepatocytes,of which baicalin had a significant effect,and the effect of cefradine on hepatocytes was increased when used in combination with cefradine.

Efficacy and safety of atomized inhalation for Shuanghuanglian on chronic pharyngitis:a Meta-analysis of the randomized controlled trials

Objective: As a Chinese drugs preparation to clear heat and remove toxicity, Shuanghuanglian (SHL) has been widely used, but the clinical efficacy and safety of SHL on chronic pharyngitis remain unclear, especially the application of atomized inhalation of SHL need to be verified. The aim of this study was to evaluate its clinical efficacy and safety for chronic pharyngitis. Methods: From the Cochrane Library, Pubmed, EMbase, Wanfang Datebase, CNKI (China National Knowledge Infrastructure), VIP (Chinese Scientifc Journals Database), CBM (Chinese Biomedicine Database), We got the qualified randomized controlled trials(RCTs) and adopted RevMan5.3 software and the risk of bias tool according to the Cochrane to perform a metaanalysis. Results: Ten RCTs, a total of 950 cases (involving 472 cases in SHL group and 478 in control group) were included. The results showed that the clinical efficacy in SHL group was superior to control group [Chi2=5.61, 95%CI (4.29, 11.43), P<0.05], and four literatures reported no obvious adverse reactions in SHL group. Conclusion: Application of atomized inhalation of SHL may have a potential advantage in treating chronic pharyngitis. However, due to the limitation of the quality and sample size of those studies, the accuracy of the result should be treated with caution, higher standardized researches are required to justify the conclusion.

Study on pharmacodynamic mechanism of compound Shuanghuanglian in prevention and treatment of pneumonia based on network pharmacology and association analysis

Background:Platform for prediction and analysis of Chinese medicine against coronavirus disease 2019(TCMAntiCOVID-19 V1.0)was used to explore the active ingredients,key targets and mechanisms of the compound coronavirus disease 2019(COVID-19),It will provide a reference for the clinical application of this prescription as a broad-spectrum agent in the prevention and treatment of pneumonia.It will also win valuable time for finding symptomatic Chinese medicine prescriptions and vaccine research and development.Methods:Set Banxia Tianma Baizhu decoction as the negative control.Qingfei Paidu decoction as the positive control,and use TCMAntiCOVID-19 V1.0 platform to predict the potential efficacy of compound Shuanghuanglian.We used the quantitative evaluation algorithm of multi-target drugs for the network disturbance of disease,by comparing the changes of network topological characteristics before and after drug intervention,and using the disturbance rate to evaluate the drug’s dryness for disease prediction.Using batman-TCM,the credibility card value of the target sets 20,then the target of traditional Chinese medicine composition is predicted,the heterogeneous network of traditional Chinese medicine composition-drug target-disease target is established.The interaction of related network is realized by JavaScript Visualization is realized,and cycloscape is edited.The average connectivity,the average shortest path length,the centrality of connectivity and the centrality of network compactness are calculated by using R’s iGraph package,and nulldistribution is used as the overall distribution to correct the disturbance rate of drugs to the real network,so as to evaluate the stability of network topology and explore its mechanism.Results:The key targets of COVID-19 were Aif1,Ccl2,Cdc20,Cxcl13,Fcer1g,Ido1,Ifng,Il10,Il1rnIl6,Ncf4,Ptger4,Spi1,Tnf,Xcl1;after the intervention,the average connectivity,the average shortest path length,the network connectivity centrality and the network tightness centrality were(2.31e+1),(2.41e+0),(6.21e−1),(1.68e−2)respectively;the total scores of network stability evaluation of drug intervention diseases were 18.29,12.59 and 19.10 respectively.Conclusion:Compound Shuanghuanglian can effectively break the stability of the disease network of COVID-19,and the overall effect of prevention and treatment of COVID-19 is close to that of the positive control Qingfei Paidu decoction,which is significantly better than that of the negative control Banxia Tianma Baizhu decoction.That is because compound Shuanghuanglian mainly acts on Aif1,Ccl2,Cdc20,Cxcl13,Fcer1g and other key targets.Compound Shuanghuanglian plays important role of inhibiting the inflammatory response of patients with COVID-19 and alleviating lung injury,so as to achieve the purpose of treating COVID-19.At the same time,in the initial stage of COVID-19 and other sudden infectious pneumoniausing compound Shuanghuanglian can win valuable time before finding symptomatic Chinese medicine prescription and vaccine research.

Bioassay-based dissolution test of Shuanghuanglian tablet

It has been difficult to perform dissolution test on solid preparations of traditional Chinese medicines （TCMs）. TCMs are different from chemical drugs in that their chemical compositions are complicated. The measurement method based on chemical approach alone is incomplete. In order to solve this problem, in this study a bioassay-based dissolution test was developed. Microcalorimetry was used to obtain growth power-time curves and biothermodynamic parameters of Staphylococcus aureus inhibited by the solution of ShuangHuangLian （SHL） tablet, which was dissolved in phosphate buffer （pH 6.8） for different times. The results of the bioassay-based dissolution test of SHL tablet demonstrated that the bioassay method might be a promising alternative for its quality control.

Effects of Shuanghuanglian oral liquids on patients with COVID-19:a randomized,open-label,parallel-controlled,multicenter clinical trial

We conducted a randomized,open-label,parallel-controlled,multicenter trial on the use of Shuanghuanglian(SHL),a traditional Chinese patent medicine,in treating cases of COVID-19.A total of 176 patients received SHL by three doses(56 in low dose,61 in middle dose,and 59 in high dose)in addition to standard care.The control group was composed of 59 patients who received standard therapy alone.Treatment with SHL was not associated with a difference from standard care in the time to disease recovery.Patients with 14-day SHL treatment had significantly higher rate in negative conversion of SARS-CoV-2 in nucleic acid swab tests than the patients from the control group(93.4%vs.73.9%,P=0.006).Analysis of chest computed tomography images showed that treatment with high-dose SHL significantly promoted absorption of inflammatory focus of pneumonia,which was evaluated by density reduction of inflammatory focus from baseline,at day 7(mean difference(95%CI),−46.39(−86.83 to−5.94)HU;P=0.025)and day 14(mean difference(95%CI),−74.21(−133.35 to−15.08)HU;P=0.014).No serious adverse events occurred in the SHL groups.This study illustrated that SHL in combination with standard care was safe and partially effective for the treatment of COVID-19.

Baicalin and rutin are major constituents in Shuanghuanglian injection involving anaphylactoid reaction

OBJECTIVE: To identify the constituents in Shuanghuanglian injection(SHLI) that correlate with anaphylactoid reaction.METHODS: Chemical fingerprints of 10 batches SHLI samples were determined by High Performance Liquid Chromatography(HPLC), and further investigated by similarity analysis. Combined with optical microscopy, both anaphylactoid experiments and confirmatory assay were displayed in Rat basophil leukemia cells(RBL-2H3) to obtain the histamine release inducing by SHLI. The content of histamine was tested by Enzyme-Linked Immuno Sorbent As-say method. Partial least squares regression(PLSR)method and HPLC-DAD-ESI-MSntechnology were conducted to analyze constituents in SHLI involving anaphylactoid reaction.RESULTS: The results of spectrum and effect relationships showed that the eight constituents were positively correlated with anaphylactoid reaction.Among which, nearly 90% of them were identified as baicalin and rutin with PLSR and HPLC-DAD-ESI-MSn. This result was in accordance with confirmatory assay on RBL-2H3 cells.CONCLUSION: Baicalin and rutin from SHLI were the main constituents involving anaphylactoid reaction.

Determination of chlorogenic acid in traditional Chinese prescription Shuanghuanglian capsule using quantitative nuclear magnetic resonance spectroscopy in combination with solid phase extraction

The determination method of chlorogenic acid in traditional Chinese prescription Shuanghuanglian capsule was established by using quantitative nuclear magnetic resonance spectroscopy(q NMR) in combination with solid phase extraction(SPE). As the capsule’s main active component, chlorogenic acid comes from the extraction of Chinese herb medicine Flos Lonicerae. The chlorogenic acid in capsule was ultrasonically extracted at room temperature using pure water as solvent. The extracting solution was enriched and cleaned using HC-C18 SPE cartridge. The effect of ultrasonic extraction, sample pretreatment conditions via SPE and q NMR experimental conditions were investigated. The q NMR experiment conditions were selected using deuterated DMSO as solvent, calibrated 1,4-phthalaldehyde as internal standard, and P1(pulse width) = 14.4 μs, d1(pulse delay time) = 1 s, NS(number of scan) = 512. The 1 H NMR peaks of δ 6.138–6.182(H-8’, d, 1 H) of chlorogenic acid was chosen as the quantitative peaks. Method validation was performed, including precision(the intra-day RSD = 1.2% and the inter-day RSD = 1.5%), linearity(correlation coefficient r>0.9999), LOD(0.0017 mg/g) and LOQ(0.079 mg/g). The recovery of the SPE-q NMR was within the range of 100.2%–103.2%. The result showed that the method was stable, accurate and reliabile. Determined by the method, the chlorogenic acid in a real Shuanghuanglian capsule was within the range of 9.68–10.35 mg/g.

Intraprostatic Injection of Shuanghuanglian in Treatment of Chronic Prostatitis (Report of 30 Cases)

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ClinicalandExperimentalStudyonTreatmentofAcuteRespiratoryTractInfectionwithShuanghuanglianAerosol（双黄连气雾剂）

Two hundred and two cases of acute respiratory tract infection ( ARI) were treated withShuanghuanglian (SHL) aerosol. In these cases, the majority were virus infection and 64% of them wascaused by the respiratory syncytial virus (RSV) . The virostatic and bacteriostatic test were done in vitro bythe cell culture method and it was shown that SHL could inhibit the RSV, para-influenza virus I - IV and 23kinds of pathogenic bacteria such as Staphylococcus aureus etc. The bacteriostatic effect was positively cor-related to the SHL concentration. Experimental study showed that SHL could enhance the NK cell activity,promote the production of a-interferon and raise the rate of lymphocyte transformation. The controlled obser-vation on SHL preparation with various dosage-forms revealed that the SHL aerosol in treating early ARlshowed better results than that of injections and oral liquid symptomatologically ( P < 0 . 05 ) . The total effec-tive rate was 96% .

Clinical Observation on Shuanghuanglian Powder (双黄连粉剂) in Treating Viral Myocarditis of Children

Objective: To study immune function of children viral myocarditis and to evaluate the clinical effect of Shuanghuanglian Powder (SHLP) by injection. Methods: The 62 patients of viral myocarditis were divided into two groups randomly, the SHLP group (n=32) treated with conventional therapy plus SHLP and the conventional treatment group (n=30) with conventional therapy alone. Their serum antibody of Coxsackie virus group B (COXB-IgM), T-lymphocyte subsets including CD+3, CD+4, CD+8 and CD+4/CD+8 were determined with ELISA and indirect immunofluorescent assay. Results: COXB-IgM was positive in 39 of the 62 patients, which was significantly different with those of normal controls (P<0.001). Patients' serum level of CD+4 cells and CD+4/CD+8 ratio decreased while CD+8 increased. After treatment, the recovery of symptoms, signs and immune function in patients of the SHLP group were significantly better than those in patients treated with conventional treatment alone (P<0.01). Conclusion: Immunoregulatory disturbance is involved in children with viral myocarditis and SHLP is an effective drug in the treatment of children viral myocarditis

双黄连口服液质量一致性评价研究

目的:探索建立双黄连口服液(SHLO)质量一致性评价方法,并对常见市售8个厂家的产品进行质量分级。方法:建立SHLO中6个指标成分的高效液相色谱含量测定方法,从同厂家不同批次、不同厂家角度分析样品含量均一性;用3个质量一致性参数[批内一致性差异(PA)、批间一致性差异(PB)、指纹图谱相似率(PC)]表征不同厂家产品质量一致性水平;运用主成分分析(PCA)模型提取一致性区分因子(P),实现对8个厂家样品质量一致性分级。结果:建立的高效液相色谱含量测定方法简便且方法学验证合格;40批样品中绿原酸、黄芩苷、连翘苷的质量浓度分别为0.66~1.25、12.71~21.89、0.39~0.66 mg·mL^(–1),均符合《中华人民共和国药典》(以下简称《中国药典》)2020年版对该制剂的限度要求,非《中国药典》2020年版指标成分新绿原酸、连翘酯苷A、汉黄芩苷的质量浓度分别为0.75~1.44、0.07~0.94、0.001 3~1.930 0 mg·mL^(–1),且同厂家样品均一性较好、不同厂家样品存在一定差异;8个厂家样品PA为1.2%~7.7%、PB为16.6%~39.1%、PC为99.3%~99.9%,依据P可将8个生产厂家样品分为3类,其中厂家F、R产品的一致性较好。结论:建立了简便的SHLO多成分定量方法,结合3个一致性评价参数和PCA模型可对不同生产厂家样品质量进行区分,数据结果可为各生产厂家质量标准提升提供参考。

复方双黄连制剂对单核巨噬细胞(RAW264.7)Fc/C3b受体活性和分泌功能的影响

【目的】探究复方双黄连制剂对巨噬细胞吞噬能力和分泌功能的影响,为复方双黄连的深度开发及畜禽用药的合理选择提供科学依据。【方法】将金银花、黄芩、连翘、穿心莲干燥粉碎后分别制备浸膏,按照比例制备双黄连口服液(金银花∶黄芩∶连翘=1∶1∶2)、复方双黄连口服液(金银花∶黄芩∶连翘∶穿心莲=1∶1∶2∶2)及穿心莲口服液,调节pH为7.0,生药浓度为1 g/mL。3种药物作用于小鼠单核巨噬细胞RAW264.7,采用CCK-8法检测细胞活力,确定3种药物安全浓度,将药物最大安全浓度以二倍稀释法稀释为3个浓度;采用脂多糖(LPS)和氢化考的松琥珀酸钠(HCSS)作用于RAW264.7细胞模拟机体炎症和免疫抑制状态,3种药物作用于这2种状态的细胞和正常细胞,采用YC、EA玫瑰花环法及中性红吞噬法检测巨噬细胞Fc/C3b受体活性和吞噬能力,采用ELISA法检测巨噬细胞分泌能力。【结果】复方双黄连、双黄连、穿心莲的安全浓度分别为0.625~2.5、3.125~12.5和0.625~2.5 mg/mL。不同浓度复方双黄连、双黄连、穿心莲作用于巨噬细胞,细胞吞噬能力均显著高于空白对照组(P<0.05)。复方双黄连可降低LPS巨噬细胞RAW264.7活跃的吞噬能力,增强HCSS巨噬细胞RAW264.7吞噬能力;不同浓度复方双黄连可使活化的巨噬细胞Fc/C3b受体活性下降,低下的Fc/C3b受体活性增强;不同浓度复方双黄连具有促进巨噬细胞分泌一氧化氮(NO)、肿瘤坏死因子-α(TNF-α)、γ-干扰素(IFN-γ)及溶菌酶(LZM)的作用,抑制LPS巨噬细胞NO、TNF-α、IFN-γ及白细胞介素-6(IL-6)的分泌。【结论】复方双黄连可通过活化巨噬细胞Fc/C3b受体活性以增强机体免疫力和抗炎能力,同时对已活化的巨噬细胞分泌功能起到抑制作用,以减少由于免疫功能亢进造成的机体损伤。研究结果为中兽药复方双黄连的深度开发提供了参考依据。