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.

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.

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.

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.

双黄连注射剂中黄芩素致敏家兔后血清中特异性抗体的测定

目的利用酶联免疫吸附测定法(ELISA)法检测双黄连注射剂中黄芩素致敏家兔后血清中特异性抗体的效价。方法将正常家兔血清与双黄连注射剂中疑似小分子半抗原物质黄芩素在体外一起孵育从而制备所需完全抗原,随后,利用这些完全抗原免疫家兔以期获得特异性抗体。运用ELISA法检测所得抗血清的抗体效价。通过间接竞争ELISA法,进一步测定血清特异性并绘制相关浓度抑制曲线。此外,采用兔免疫球蛋白E(IgE)ELISA试剂盒检测各组家兔抗血清中相对应的IgE抗体含量。结果与阴性对照组比较黄芩素的抗体效价(OD)值是其2倍,表明其具有潜在的致敏性。通过整理数据,绘制抑制率曲线得到回归方程I=0.138lgC+0.402,R^(2)=0.959,显示出良好的拟合度。半数抑制浓度IC_(50)=5.178 mg·mL^(-1)。此外,本实验结果明显表明外源性IgE型抗体已经在家兔体内产生,从而进一步证实了黄芩素的致敏性。结论在本实验所模拟家兔致敏全过程结果中表明黄芩素具有明显的致敏性。这一发现对于双黄连注射剂的安全使用具有重要的指导意义,也为相关药物的研发和生产提供了有价值的参考。

双黄连注射液不良反应样品筛查方法研究

目的:建立双黄连注射液不良反应样品筛查方法,为产品质量控制、临床安全用药提供参考。方法:采用鞣质检测和细胞响应谱法对双黄连注射液正常与不良反应样品进行筛查,以改良干酪素法进行鞣质检测,另选取肥大细胞RBL-2H3作为模式生物,使用xCELLigence实时细胞分析系统对双黄连注射液诱导的RBL-2H3脱颗粒效应进行了实时监测。结果:鞣质检测无法有效区分正常样品与不良反应的样品,细胞响应谱法可准确分辨不良反应样品。结论:基于肥大细胞脱颗粒效应的细胞响应谱方法操作简便、高通量、准确可靠,为双黄连注射液质量控制和临床安全用药提供参考。

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.

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

Ｉｎｔｒａｐｒｏｓｔａｔｉｃ Ｉｎｊｅｃｔｉｏｎ ｏｆ Ｓｈｕａｎｇｈｕａｎｇｌｉａｎ ｉｎ Ｔｒｅａｔｍｅｎｔ ｏｆ Ｃｈｒｏｎｉｃ Ｐｒｏｓｔａｔｉｔｉｓ （Ｒｅｐｏｒｔ ｏｆ ３０ Ｃａｓｅｓ）ＺＨＡＮＧＸｉａｎ－ｔａｏ，ＺＨＡＮＧＹｉｎｇ－ｅ...

双黄连粉针剂抗病毒作用

目的 明确双黄连粉针剂体内外抗病毒谱及作用机制。方法 采用组织细胞培养法、染料摄入法 ,检测双黄连粉针剂对不同种病毒的作用 ,改变给药时间、途径 ,探讨双黄连粉针剂抗病毒作用环节。建立病毒性心肌炎、胰腺炎、流感动物模型 ,通过组织病理学检查法 ,观察双黄连粉针剂对感染动物的保护作用。结果 双黄连粉针剂具有明显抗流感病毒、呼吸道合胞病毒、腺病毒 型、单纯疱疹病毒 及 型、柯萨奇病毒 B3 ,A1 6、新型肠道病毒71型的作用。对脊髓灰质炎病毒 型、埃可病毒 6型、麻疹病毒、水泡性口炎病毒有一定的抑制作用。并能显著抑制肺炎、心肌炎、胰腺炎的发生 ,疗效与清开灵相似 ,优于穿琥宁粉针剂、鱼腥草、莪术油注射液。结论 双黄连粉针剂是一个较广谱的抗病毒针剂 ,抗病毒作用是多途径的。

4种中药注射剂皮肤过敏试验方法与意义的探讨

黄芪多糖粉针剂皮下法皮肤过敏试验阳性率12．3％，清开灵粉针剂3．0％，清开灵注射液7．6％，双黄连注射液6．3％。青霉素过敏反应率为0．7％～10％。不同剂型（粉针与液剂），不同浓度等均可能会影响皮试阳性率。皮试阳性不用药。皮试阴性用药，用药后仍有过敏反应发生。黄芪多糖粉针剂皮试阴性，用药后过敏反应发生率为2．1％。清开灵为0．4％（1／224），双黄连注射液为0．9％～2．6％。双黄连注射液在人群中的致敏率为8．6％（31／360包括皮试阳性数加过敏反应数）。清开灵粉针剂组致敏率为4．5％（6／132），清开灵注射液组致敏率9．1％（12／132）。黄芪多糖粉针剂组致敏率15．4％（62／402）。合计致敏率10．8％（111／1026）。青霉素皮下法试验阳性主要表现以皮丘为主，伴有周围红晕，而中药注射液皮肤以红晕为主，皮丘多不明显。皮试对减少黄芪多糖粉针剂、清开灵粉针剂、双黄连注射液用药后过敏反应有一定意义。可作为临床减少不良反应的措施之一。

双黄连气雾剂与注射剂的药效学及临床应用研究

药效学实验表明：双黄连两种剂型均能增强人外周血ＮＫ细胞活性，具有促进淋巴细胞生成α－干扰素的能力，同时具有广谱抑菌、抑病毒作用。以注射剂生物利用度为１００％，气雾剂绝对生物利用度为９６．８８％（黄芩甙）和９６．６９％（绿原酸），临账观察表明：气雾剂与注射剂的显效率分别为８４％和８３％，总有效率分别为９２％和９３％。

BN大鼠和豚鼠评价双黄连注射液的过敏反应

目的为摸索适合评价中药过敏反应的动物,采用BN大鼠和豚鼠评价双黄连注射液过敏的反应。方法BN大鼠和豚鼠分为ip或iv临床等倍剂量组(分别为360及300mg.kg-1)、iv临床2倍剂量组(分别为720及600mg.kg-1)。以双黄连注射液为致敏原,分别对2种动物进行致敏,隔日1次,共3次。同时设生理盐水对照组,观察各组过敏反应症状;采用ELISA法测量血清和组织中组胺含量,HE染色法光镜下观察肺组织和气管病理改变。结果双黄连注射液可以造成BN大鼠及豚鼠发生过敏反应,过敏反应发生率BN大鼠组明显高于豚鼠组,分别为62.86%和36.11%。双黄连注射液可使BN大鼠和豚鼠血清、肺和气管中组胺含量均明显升高,与各相应的对照组比较均具有显著性差异,同时,从数据上看,BN大鼠各组的组胺释放率均高于豚鼠各组。结论双黄连注射液可造成BN大鼠及豚鼠发生过敏反应。BN大鼠的敏感性可能高于豚鼠,但还需进一步研究。

双黄连注射剂与西药配伍研究的系统评价

目的探讨研究双黄连注射剂与西药注射剂和配伍宜忌,为双黄连注射剂的临床合理使用提供依据。方法全面收集双黄连注射剂(水针和粉针)与西药注射剂配伍的实验室和临床研究,提取有关双黄连剂型与用量、配伍药物及用量、输液类型、配制液理化检测指标以及药物代谢、药效指标和不良反应等数据,按照配伍药物的类别进行分类,描述性分析其物理性状、化学成分、药物代谢和安全性等,并据此评价配伍的宜忌。结果共纳入60篇文献,其中实验研究38篇,1999～2002年间发表者占总数的50%。所有研究共涉及西药注射剂53个,并以与β内酰胺类、氨基甙类及喹诺酮类抗生素配伍的研究为多,共发现31个配伍禁忌。本系统评价结果还显示,现有实验研究指标选择片面,在今后的研究设计中应予注意。结论双黄连注射剂与西药针剂存在广泛配伍禁忌,在没有研究清楚前,不可盲目配伍使用。现有中药注射剂与其他药物配伍的基础研究还很薄弱,企业应加强组织研究,主动收集上市后研究的数据,不断完善说明书。

注射用双黄连的免疫毒性研究

目的观察绿原酸和注射用双黄连的致敏性。方法用豚鼠全身主动过敏反应试验观察注射用双黄连过敏原性;用大鼠、小鼠、豚鼠等同种、异种动物间的被动皮肤过敏反应试验,比较绿原酸和注射用双黄连过敏原性。结果注射用双黄连致敏性高于绿原酸。结论推测注射用双黄连作为一个复方引起不良反应的可能性大于绿原酸。

双黄连粉针在5种输液中的稳定性考察

本实验应用系数倍率法测定双黄连粉针与５种输液及青霉素配伍时黄芩苷、绿原酸、连翘苷的含量，并考察ｐＨ值变化。实验结果表明双黄连粉针可与其中４种输液配伍，亦可与青霉素配伍，不宜与含钾盐的复方葡萄糖注射液配伍。

双黄连粉针中黄芩甙的药动学

目的：测定双黄连粉针中主要有效成分黄芩甙的体内药代动力学参数。方法：采用高压液相色谱法，流动相０１ｍｏｌ／Ｌ磷酸二氢钾∶乙酸∶四氢呋喃（１０００∶１００∶１９０），色谱柱ＥＲＣＯＤＳ１１６１（６ｍｍ×１００ｍｍ），检测电压＋６５０ｍＶ，柱温５５℃，流速０８ｍｌ／ｍｉｎ。结果：双黄连粉针中黄芩甙在大白鼠体内药代动力学参数是Ｖｃ＝０．０９Ｌ，Ｋｅ＝３．７４／ｈ，Ｋ１２＝２．５６／ｈ，Ｋ２１＝１．０４／ｈ，ｔ＝０．２２ｈ，α＝６．７７ｎｇ／（ｍｌ·ｈ），β＝０．４５ｎｇ／（ｍｌ·ｈ），ＡＵＣ＝１１．２（μｇ·ｈ）／ｍｌ。结论：双黄连粉中黄苓甙在大白鼠体内呈二室模型分布。

双黄连注射剂不良反应文献评价

目的了解双黄连注射剂不良反应(ADR)在不同年龄、系统和严重程度的分布、构成及影响因素,为双黄连注射剂临床合理应用和上市后再评价提供参考。方法计算机检索中国期刊全文数据库(CNKI,1979.1～2009.9)、中国科技期刊全文数据库(VIP,1989.1～2009.9)和中国生物医学文献数据库(CBM,1978.1～2009.9),按纳入与排除标准选择文献、提取资料,并按不同类别(年龄、过敏史、涉及系统、联合用药等)计算出双黄连注射剂ADR总数、构成比,根据临床研究中可获得的用药人数与ADR病例数估算ADR发生率。结果①共纳入452篇文献,合计2799例ADR病例,近20年发表文献以病例报告为主(84.51%),根据69篇文献报告的31165例用药人数和1013例ADR数,计算出ADR总发生率为3.25%。②ADR病例中男女比例为1.13?1,18岁以下未成年人占32.71%。③ADR病例药物过敏史以青霉素最多(占13.38%),其次为磺胺、头孢菌素、喹诺酮类等。④ADR病例原患疾病中呼吸系统占91.75%,其次为消化系统(5.17%)、泌尿系统(1.11%)等。⑤药物联用以青霉素最多,发生ADR的风险是单用的3.14倍[95%CI(2.58,3.81)]。⑥ADR涉及全身多个系统和器官,构成比由高到低依次为皮肤、消化系统、全身反应、呼吸系统、神经系统、心血管系统、局部反应、泌尿系统、血液系统及其它。⑦按WHO药物ADR严重程度分级,Ⅰ、Ⅱ、Ⅲ、Ⅳ级分别占6.36%、5.48%、45.62%、2.12%;52.42%的ADR病例未描述具体临床表现与转归。⑧12例死亡病例除1例因疼痛诱发心肌梗死致死外,其余均因过敏性休克致死,2例曾合并用药,ADR出现最快者1分钟内。⑨双黄连粉针剂ADR发生率(2.25%)低于注射液(4.14%),其差异有统计学意义。结论双黄连注射剂ADR临床表现以皮肤过敏和消化道反应为主,联合用药会增加ADR风险,粉针剂比注射液安全性高。建议加强对双黄连注射剂从生产到临床应用过程的监管与监测,严格规范ADR报告,分级风险评估与管理ADR。

双黄连注射剂与输液配制后微粒变化的系统评价

目的探讨双黄连注射剂与输液配制后微粒变化的情况,为合理选择溶媒提供参考。方法全面收集双黄连注射剂与输液配伍的相关研究,评价纳入研究的报告质量,并提取相关数据进行系统分析。结果共检出338个条目,筛选后纳入24个研究。文献质量评价结果显示,纳入研究的质量不高,结果差异较大。总体上双黄连注射剂与输液配制后微粒增多,且不同厂家、同一厂家的不同批次及不同输液间存在差异较大。溶媒pH值对微粒影响较大,与浓度和放置时间也有一定关系。双黄连注射剂与生理盐水配制后微粒少于与5%GS、10%GS及其他溶媒配制。结论双黄连注射剂与生理盐水配制后微粒最少,一般情况下可为首选溶媒。临床合理用药相关因素值得进一步研究,同时要注意研究方案的科学性。

双黄连粉针剂中黄芩苷的高效毛细管电泳-电导法测定

建立了双黄连粉针剂中黄芩苷含量测定的高效毛细管电泳电导法 ;以未涂层融硅毛细管 (75μm×50cm)为分离柱 ,Tris-H3BO3 为电泳介质 (15mmol/LTris ,1.5mmol/LH3BO3) ,在18kV、pH8.5的碱性环境下 ,柱端电导检测双黄连粉针剂中的黄芩苷含量 ;重点探讨了缓冲溶液的种类、浓度、分离电压、进样时间对检测的影响 ;该法的线性范围为10～720mg/L,检出限为2.5mg/L;结果表明该法简便、快速、结果准确 ,适用于含黄芩苷的制剂含量的测定。

双黄连注射剂过敏症动物模型的研究

目的:探索双黄连注射剂中引起Ⅰ型过敏反应的致敏性物质,建立双黄连注射剂过敏症动物模型。方法:将双黄连注射剂与正常兔血清体外孵育制备抗原,免疫新西兰纯种兔,制备抗双黄连注射剂中过敏性物质的抗血清,并采用ELISA法检测其效价及抗体特异性。结果:双抗夹心ELISA检测抗血清效价,稀释至1:128倍仍为阳性,间接竞争ELISA测定血清特异性并绘制抑制曲线,得到回归方程I=13.362lgC+50.503,相关系数r=0.966,半数抑制浓度IC50=0.92mg/mL。结论:此法初步建立了双黄连注射剂过敏症动物模型,对双黄连注射剂中致敏物质的研究有一定意义。