复方苦参注射液胸腔灌注治疗乳腺癌致恶性胸腔积液的疗效及安全性观察

目的:探讨复方苦参注射液胸腔灌注治疗乳腺癌致恶性胸腔积液的临床疗效和安全性。方法:选择2019年9月~2023年2月在某院乳腺甲状腺外科就诊的31例乳腺癌致恶性胸腔积液患者为研究对象,采用随机数字表法分为观察组(n=16)和对照组(n=15)。两组均行胸腔闭式引流术并给予地塞米松磷酸钠注射液胸腔灌注,观察组在此基础上给予复方苦参注射液,对照组给予顺铂注射液。比较两组临床疗效、胸腔积液中肿瘤标记物[癌胚抗原(CEA)、糖类抗原153(CA153)、糖类抗原125(CA125)]含量以及不良反应发生情况。结果:观察组、对照组的疾病控制率分别为75.00%和86.67%(P>0.05),无统计学差异。两组治疗前后胸腔积液中肿瘤标记物CEA、CA153、CA125含量均较治疗前下降,两组肿瘤标记物下降情况无统计学差异(P>0.05)。但治疗过程中,观察组胸痛、乏力的发生率均低于对照组(P<0.05)。结论:复方苦参注射液胸腔灌注治疗乳腺癌致恶性胸腔积液的临床疗效与单用顺铂疗效相当,且治疗过程中胸痛及乏力的发生率更低。

Compound Kushen injection induces immediate hypersensitivity reaction through promoting the production of platelet-activating factor via de novo pathway

OBJECTIVE Compound Kushen injection(CKI)is a bis-herbal formulation extracted from Kushen(Radix Sophorae Flavescentis)and Baituling(Rhizoma Heterosmilacis Japonicae).Clinically,it is used as the adjuvant treatment of cancer.However,with the increased application,the cases of immediate hypersensitivity reactions(IHRs)also gradually rise.In this study,we investigated the underlying mechanism(s)and active constituent(s)for CKI-induced IHRs in experimental models.METHODS T helper 2(Th2)immunity-amplified mice were prepared by aluminum adjuvant.Anaphylactic shock was detected by measuring rectal thermometry in propranolol pretreated mice.For evaluating microvascular permeability,Evans blue extravasation assay was used.Platelet-activating factor(PAF),serum total IgE(tIgE)and mouse mast cell protease 1(MMCP1)were measured by ELISA.RESULTS The obtained results showed that CKI did not elevate serum tIgE and MMCP1 after consecutive immunization for five weeks,but could induce Evans blue extravasation(local)and cause obvious hypothermia(systemic)after a single injection.Further study showed that alkaloids in Kushen,especially matrine,were responsible for CKI-induced IHRs.Mechanism study showed that various PAF receptor antagonists could significantly counter CKI-induced IHRs locally or systemically.In cell system,CKI was able to promote PAF production in a non-cell-selective manner.In cell lysate,the effect of CKI on PAF production became stronger and could be abolished by blocking de novo pathway.CONCLUSION In conclusion,our study identifies,for the first time,that CKI is a PAF inducer.It causes non-immunologic IHRs,rather than IgE-dependent IHRs,by promoting PAF production through de novo pathway.Alkaloids in Kushen,especially matrine,are the prime culprits for IHRs.Our findings may provide a potential approach for preventing and treating CKI-induced IHRs.

苦参不同组织中生物碱类成分的LC-MS分析

目的 通过对苦参Sophora flavescens不同组织样品中生物碱类成分的LC-MS分析,探讨苦参中生物碱类成分的质谱裂解规律及其在不同样品中的异同。方法 首先利用UPLC-Q-Exactive-OrbitrapMS对5年生苦参根中的生物碱类成分进行全面的鉴定分析。再利用UPLC-QQQ-MS/MS技术,对苦参无菌幼苗不同组织部位(根、茎、叶)、苦参愈伤组织中的生物碱类成分进行定量分析,比较各样品中生物碱类成分的异同。结果 从5年生苦参根中共鉴定出47个生物碱类成分;利用UPLC-QQQ-MS/MS对其中17个主要的生物碱类成分进行了定量分析,发现不同苦参样品中生物碱类成分差异明显,其中苦参无菌幼苗各组织部位中生物碱的含量相对较高。结论 建立了一种苦参生物碱类成分快速定性定量分析方法,从5年生苦参根中快速鉴定出47种生物碱;通过对苦参不同组织样品中17个生物碱类成分的定量分析,为后续利用比较转录组测序技术,挖掘并鉴定苦参中生物碱类成分生物合成关键酶的研究奠定基础。

ICP-MS法测定复方苦参注射液中铜、砷、镉、汞、铅元素的残留量

目的:建立复方苦参注射液中铜、砷、镉、汞、铅5种元素残留量的测定方法。方法:样品经消解后采用电感耦合等离子体质谱法(ICP-MS法)测定,射频功率为1 550 W,载气(高纯氩气)流速为1.05L·min^(-1),等离子气体流速为15.0 L·min^(-1),蠕动泵转速0.2 r·s^(-1),采样深度10 mm,重复次数为3次。通过在线加入内标锗、铟、铋元素的方法来校正基体效应和干扰。结果:各元素在各自的检测质量浓度范围内线性关系良好(r>0.999 8),进样精密度RSD在0.8%～1.5%,重复性RSD在1.8%～6.4%,5种元素高、中、低3种浓度的加样回收率在78.9%～106.4%,样品在消解后48 h内各元素稳定性良好,铜、砷、镉、汞、铅检测下限分别为24.04、35.92、4.84、5.43、16.18 pg·mL^(-1),定量下限分别为80.13、119.72、16.13、18.10、53.94 pg·mL^(-1)。10批样品中铜含量范围为0～17.16 ng·mL^(-1),砷含量范围为6.07～11.04 ng·mL^(-1),镉含量范围为0.04～0.71 ng·mL^(-1),汞含量范围为0～1.14 ng·mL^(-1),铅含量范围为2.43～12.43 ng·mL^(-1),5种元素含量均在合格范围内。结论:本方法可用于复方苦参注射液中铜、砷、镉、汞、铅5种元素的含量测定。

复方苦参注射液UPLC指纹图谱检测及6种成分含量测定

目的建立复方苦参注射液的超高效液相色谱法(UPLC)指纹图谱,并同时测定其中6种指标成分的含量。方法采用ACQUITY UPLC CSH^(TM)C_(18)(2.1 mm×100 mm,1.7μm)色谱柱,以甲醇2 g·L^(-1)磷酸二氢钾为流动相梯度洗脱,流速0.3 mL·min^(-1),柱温30℃,检测波长211 nm。结果建立复方苦参注射液UPLC指纹图谱,确立了9个共有峰,10批样品相似度均大于0.99。甲基氧化偶氮甲醇樱草糖苷、苦参碱、槐果碱、槐定碱、氧化槐果碱、氧化苦参碱6种成分在相应浓度范围内与峰面积的线性关系良好,平均加样回收率分别为100.42%、100.90%、101.45%、103.85%、99.95%、100.46%,相对标准偏差(RSD)值分别为1.96%、0.67%、1.60%、1.15%、1.02%、1.20%。结论本研究建立的复方苦参注射液的UPLC指纹图谱及多成分含量测定方法,操作简单,重复性好,稳定可靠,为复方苦参注射液的质量标准研究提供更加全面的依据。

HPLC法同时测定复方苦参注射液中7个成分含量及指纹图谱研究

目的:建立HPLC法同时测定复方苦参注射液中7个成分(甲基氧化偶氮甲醇樱草糖苷、苦参碱、槐果碱、槐定碱、氧化槐果碱、氧化苦参碱、番石榴酸)的含量及指纹图谱研究,为其质量控制提供依据。方法:采用Waters Xselect CSH^(TM)C_(18)色谱柱(250 mm×4.6 mm, 5μm),以甲醇-0.2%磷酸二氢钾为流动相,梯度洗脱,流速0.6 mL·min^(-1),柱温30℃,检测波长211 nm。结果:在建立的液相色谱条件下,测定的甲基氧化偶氮甲醇樱草糖苷、苦参碱、槐果碱、槐定碱、氧化槐果碱、氧化苦参碱、番石榴酸7个成分的平均加样回收率分别为95.7%、101.3%、100.8%、101.7%、102.6%、102.5%、99.5%,RSD分别为2.0%、0.72%、0.90%、0.74%、1.4%、0.96%、1.8%;10批次样品中7个成分的含量分别为0.53~0.73、2.66~4.22、0.75~1.11、0.70~0.89、2.20~2.84、7.62~9.95、1.63~2.20 mg·mL^(-1)。在相同液相色谱条件下建立的指纹图谱测定方法确立了9个共有峰,生成对照图谱,10批样品的相似度测定结果在0.99以上。结论:本研究同时对复方苦参注射液的主要成分进行定量分析和指纹图谱进行定性分析,含量测定准确,指纹峰特征性强,该方法稳定可靠,可为复方苦参注射液的质量控制与评价提供参考。

HPLC-ELSD检测复方苦参注射液中大分子物质

目的建立高效液相凝胶色谱法(HPLC-ELSD)对复方苦参注射液中大分子物质进行检测。方法采用TSKgel G2000SWXL(7.8 mm×300 mm,5μm)色谱柱,20 mmol·L^-1醋酸铵水溶液为流动相,柱温25℃,流速0.8 mL·min^-1,蒸发光检测器参数(漂移管温度55℃、雾化温度55℃、氮气流速1.8 L·min^-1),建立大分子物质检测方法,并对11批复方苦参注射液进行大分子物质检测。结果11批复方苦参注射液中均未检出大分子物质,说明经过生产过程的逐步除杂,大分子物质在复方苦参注射液成品中已去除。结论此方法简便、快捷,可用于复方苦参注射液中大分子物质的检测,可为复方苦参注射液质量标准提高提供依据。

Mechanisms of Compound Kushen Injection for the treatment of bladder cancer based on bioinformatics and network pharmacology with experimental validation

Bladder cancer is the most common malignancy of the urinary system.Compound Kushen Injection(CKI)is a Chinese medicinal preparation that has been widely used in the treatment of various types of cancers in the past two decades.However,the pharmacological effect of CKI on bladder cancer is not still completely understood.In the current study,network pharmacology com-bined with bioinformatics was used to elucidate the therapeutic mechanism and potential targets of CKI in bladder cancer.The mech-anism by which CKI was effective against bladder cancer was further verified in vitro using human bladder cancer cell line T24.Net-work pharmacology analysis identified 35 active compounds and 268 target genes of CKI.Bioinformatics data indicated 5500 differen-tially expressed genes associated with bladder cancer.Common genes of CKI and bladder cancer suggested that CKI exerted anti-blad-der cancer effects by regulating genes such as MMP-9,JUN,EGFR,and ERK1.Functional enrichment analysis indicated that CKI ex-erted therapeutic effects on bladder cancer by regulating certain biological processes,including cell proliferation,cell migration,and cell apoptosis.In addition,Kyoto Encyclopedia of Genes and Genomes enrichment analysis implicated pathways related to cancer,bladder cancer,and the PI3K-Akt signaling pathway.Consistently,cell experiments indicated that CKI inhibited the proliferation and migration of T24 cells,and induced their apoptosis.Moreover,RT-qPCR and Western blot results demonstrated that CKI was likely to treat bladder cancer by down-regulating the gene and protein expression of MMP-9,JUN,EGFR,and ERK1.CKI inhibited the prolif-eration and migration,and induced the apoptosis of T24 bladder cancer cells through multiple biological pathways and targets.CKI also exhibited significant effects on the regulation of key genes and proteins associated with bladder cancer.Overall,our findings provide solid evidence and deepen current understanding of the therapeutic effects of CKI for bladder cancer,and further support its clinical use.