Cardioprotective effect of Yiqi Huoxue granule through regulation of mitophagy after myocardial infarction in rats

Objective:To investigate the cardioprotective effect of Yiqi Huoxue granule(YQHXG)in the regulation of autophagy in rats induced with myocardial infarction(MI).Methods:An acute MI animal model was established by ligation of the left anterior descending branch of the coronary artery in Sprague-Dawley rats.Besides,20 rats received sham operation were classified into a control group.The remaining 59 rats were randomly divided into MI model group(n=19),YQHXG group(n=20),and perindopril group(n=20).Relevant indicators on days 7 and 28 were observed in each group.Left ventricular function was determined by echocardiography.The structure and morphology of mitochondria,and the number of autophagic vesicles,were observed by transmission electron microscopy.The mRNA and protein expression levels of LC3,FUNDC1,Beclin-1,and BNIP3 were examined in the tissue of the MI marginal area.Results:Compared with the MI model group,YQHXG showed obvious improvements in cardiac functions.Observing the microscopic morphology of the heart tissue,myocardial tissue damage attenuated,autophagic signs of autophagosomes and autolysosomes reduced,vacuolization in mitochondria mitigated,and mitochondria arranged in order.YQHXG could reduce the degree of tissue lesion after MI and regulate the expression of autophagy-related molecules at different stages.On Day 7,YQHXG significantly downregulated the expression of Fundc1,Becn1,Bnip3 mRNA and reduced the levels of FUNDC1,Beclin-1,BNIP3,and LC3 B proteins expression(all P<.001).On Day 28,YQHXG could upregulate the expression of Becn1,Fundc1 and Bnip3 mRNA and increased the levels of the corresponding proteins expression(all P<.001).Besides,it also increased LC3 B protein expression level(P=.0344).Conclusion:YQHXG regulated the expression of mitochondrial autophagy-related factors in myocardial tissue and mitochondrial autophagic activity at different stages to protect the heart following MI.

Effect of Qinggan Huayu granule on H22 liver ascitic tumor mice

Objective:To evaluate the therapeutic effect of Qinggan Huayu granule on mice with H22 liver cancer ascites tumor.Methods:A H22 liver cancer ascites mouse model was established by intraperitoneally injecting H22 liver cancer cells.Mice were randomly divided into the model group,the Ganfule group(1.35 g/kg),the fluorouracil group(50 mg/kg i.p),the Qinggan Huayu granule groups at low(0.67 g/kg),medium(1.34 g/kg),and high(2.68 g/kg)doses.Then the mice were administered continuously for 10 days and body weight and abdominal circumference were monitored every 3 days.On day 11,eight rats in each group were randomly selected for dissection to detect the amount of peritoneal water,peritoneal permeability and histopathological changes.The remaining mice were observed for survival.In addition,the vascular endothelial growth factor A(VEGFA)and vascular endothelial growth factor receptor 2(VEGFR2)were determined by Western blotting.Results:Compared with the model group,the weight growth of mice in the fluorouracil group and the medium-dose and high-dose Qinggan Huayu granule groups was slower(P<0.05).Moreover,the abdominal circumference of mice in each treatment group was increased slowly.There were significant differences in abdominal circumference between the fluorouracil group,the medium-dose group and the control group from day 6(P<0.05)while the abdominal circumference of the high dose group was significantly smaller than that of the control group from day 12(P<0.05).Moreover,compared with the model group,the amount of ascites in the medium-and high-dose Qinggan Huayu granule groups was decreased significantly(P<0.05).The optical density value of ascites supernatant in medium-and high-dose Qinggan Huayu granule group and the fluorouracil group decreased significantly(P<0.05)and the survival period of the medium-dose Qinggan Huayu granule group and the fluorouracil group was prolonged prominently(P<0.05).There was no significant difference in the low-dose Qinggan Huayu granule group and the Ganfule group.Peritoneal histopathological assay showed more complete peritoneal structure,less edema and less angiogenesis of the peritoneum in the fluorouracil group and the medium-and high-dose Qinggan Huayu granule group,which was better than that of the Ganfule group and the low-dose group.Compared with the model group,the expressions of VEGFA and VEGFR2 in the medium-dose Qinggan Huayu granule group decreased significantly(P<0.05,P<0.01).Conclusion:Qinggan Huayu granule can inhibit ascites production in the mice model with H22 liver cancer ascites tumor,prolong the survival of mice,and reduce peritoneal permeability and suppress the increase of peritoneal neovascularization.The mechanism may be related to the inhibition of VEGF/VEGFR pathway.

Effect of Yiqi Huoxue (益气活血) Herbs on Vascular Endothelial Cells and Platelets in Patients with Chronic Cor Pulmonale

Objective: To observe the clinical effect of Yiqi Huoxue (益气活血, YQHX) herbs in treating the patients with chronic cor pulmonale and to explore its mechanism by determining the relationship of oxida-tion/antioxidation system and how such herbs change on the function of endothelial cells and platelets. Methods: Fifty-eight patients were divided into two groups: conventional therapy group (control group, 28 patients) and convention plus YQHX herbs group (treated group, 30 patients). The control group received conventional management. The treated group were treated with YQHX 150 ml, twice a day, plus the conventional treatment, and the clinical efficacy was recorded. The lipid peroxidation (LPO), superoxide dismutase (SOD), ot-granule membrane protein (GMP140), partial pressure of arterial oxygen (PaO2), partial pressure of carbon dioxide in artery (PaCO2) and circulating endothelial cells (CEC) were measured respectively before and after treatment, and the relationship between various parameters were analyzed. The results were compared with those of 10 healthy subjects got at the same period. Results: (1) The effective rate and PaO2 of the treated group was higher than that of the control group and there were no difference in PaCO2 between the two groups. (2) The levels of LPO, GMP140, CEC in all the patients before therapy were significantly higher than those of the healthy group, and there were marked decrease in the levels of those after treatment (all P<0. 01). On the contrary, the levels of SOD in all the patients before therapy were markedly lower than those in the healthy subjects and increased after treatment, P<0. 01. (3) The increase of SOD in the treated group was significantly more obvious than that of the control group. In the treated group, the decrease of LPO, GMP140, CEC were markedly more obvious than those in the control group (all P<0. 01). (4) The number of CEC, as well as GMP140, was negatively correlated to PaO2(P<0.01) and SOD (P< 0. 01), which was positively correlated to LPO (P<0. 01). There was a positive correlation between CEC and GMP140 (P<0.05). Conclusion: YQHX herbs in treating chronic cor pulmonale proved to be effective by balancing the oxidation and antioxidation, protecting the pulmonary endothelial cells and activated platelets and helpful in treating respiratory failure.

基于UPLC-Q-TOF-MS法和网络药理学探讨草归除湿活血颗粒治疗高尿酸血症的机制

目的考察草归除湿活血颗粒剂的提取工艺,并探究其通过降尿酸治疗高尿酸症的机制。方法通过HPLC测定草归除湿活血颗粒水提液中各成分的含量,筛选最佳提取条件;利用UPLC-Q-TOF-MS分析其活性成分,再将得到的活性成分结合高尿酸血症的潜在靶点,进行PPI网络构建及GO、KEGG分析。结果最佳提取工艺为10倍水浸泡0.5 h,加热回流提取2次,每次1 h。UPLC-Q-TOF-MS得221个活性成分,网络药理学分析得到183个潜在交集靶点,富集分析得到PI3K-Akt信号通路、脂质和动脉粥样硬化、HIF-1等信号通路。结论草归除湿活血颗粒可能通过调节PI3K-Akt信号通路、脂质和动脉粥样硬化、HIF-1等信号通路来发挥降尿酸的作用。

基于Box-Behnken设计-响应面法优化调肝活血稳压颗粒提取工艺及其质量标准的研究

目的优化调肝活血稳压颗粒的提取工艺,并对其质量标准进行研究。方法本研究以浸膏得率及丹酚酸B的转移率的综合评分为指标,采用单因素考察结合Box-Behnken设计-响应面法优化调肝活血稳压颗粒的提取工艺,采用薄层色谱法对方中药材进行鉴别。结果确定最终提取工艺为:第一次加入10倍量水煎煮90 min,第二次加入6倍量水煎煮60 min。测定方法结果准确可靠。薄层色谱法检测出天麻、栀子、丹参等三味中药的特征性斑点。结论优化后的提取工艺操作简单,提取效果良好,结果稳定可靠。

Effect of Yifei Huoxue Granule (益肺活血颗粒) on the Proliferation of Rat Pulmonary Artery Smooth Muscle Cells upon Exposure to Chronic Hypoxic Conditions in Vitro

Objective： To investigate the inhibitory effect of Yifei Huoxue Granule （益肺活血颗粒, YFHXG） on the hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells （PASMCs） and its mechanism of decreasing pulmonary arterial pressure. Methods： Twenty male Sprague-Dawley （SD） rats were randomly divided into four groups： saline, and 0.66, 3.30 and 16.50 g/kg of YFHXG groups, the saline and different concentrations of YFHXG were given twice daily for 7 days, respectively. Serum-pharmacology method was used in the preparation of YFHXG serum. Tissue block anchorage was employed in the primary culture of rat PASMCs. The PASMCs were randomly divided into normoxia group, hypoxia group, and hypoxia＋YFHXG group （0.66, 3.30 and 16.50 g/kg doses of YFHXG-treated serum groups, exposed to hypoxic condition）. PASMCs in normoxia and hypoxia group were cultured with saline serum, hypoxia＋YFHXG groups were cultured with different concentrations of YFHXG serum. Cell viability was assessed with 3-（4,5-dimethylthiazol-2-yl）-2,5- diphenyltetrazolium bromide （MTT） assay. Cell cycle was analyzed using flow cytometry. In addition, hypoxia inducible factor-l-alpha （HIF-1α） protein expression was evaluated by immunocytochemistry analysis, the concentration of intracellular reactive oxygen species （ROS） and Ca2＋ were determined by laser scanning confocal microscopy （LSCM）. Results： MTT assay and flow cytometry showed that hypoxia could directly activate the proliferation of PASMCs, while YFHXG dose-dependently inhibited hypoxia-induced proliferation of rat PASMCs. Immunocytochemistry showed that hypoxia enhanced HIF-1α protein expression, and LSCM showed that hypoxia significantly increased intracellular ROS and Ca2., while YFHXG decreased the expression of HIF- 1α and attenuated the hypoxia-induced increase in intracellular concentration of ROS and Ca2＋. Conclusions： YFHXG could inhibit hypoxia-induced proliferation of rat PASMCs, which may decrease pulmonary arterial pressure and vascular remodeling. The anti-hypoxia effect of YFHXG may be explained by its regulation of HIF- 1 α expression and of the levels of intracellular ROS and Ca2＋.

Effectiveness of Bushen Huoxue Granule (补肾活血颗粒) on 5-Serotonin and Norepinephrine in the Brain of Parkinson's Disease Patients with Depressive State

Objective：To observe the clinical curative effect of Chinese medicine Bushen Huoxue Granule（补肾活血颗粒,BHG） on Parkinson＇s disease（PD） patients with depressive state.Methods：Sixty-two PD patients with depressive state were randomly assigned to two groups by using a random number table,31 in each group.Madopar was given to all as the conventional treatment.The fluoxetine hydrochloride dispersible tablet was given to the patients in the control group and BHG was given to those in the treatment group.The therapeutic course for all was 12 weeks.Before and after treatment,Hamilton depression rating scale（HAMD） was applied to judge the curative effect,and the changes of cerebral neurotransmitters levels in the brain of patients were detected by encephalofluctuograph technique.Results：The scores of HAMD in the two groups were decreased markedly after 12-week treatment.It was lower in the treatment group than that in the control group with significant difference（P〈0.01）.The contents of norepinephrine（NE） and 5-serotonin（5-HT） in the PD patients were obviously lower than normal value.There was no significant difference between the two groups before treatment（P〉0.05）.The contents of NE and 5-HT were all increased in the two groups after treatment（P〈0.05）,with significant differences between the two groups（P〈0.01）.Conclusion：BHG could increase the contents of NE and 5-HT in PD patients＇ brain to improve the depressive state of PD patients.

金花清感颗粒联合布地奈德、沙丁胺醇、异丙托溴铵三联雾化吸入疗法在儿童肺炎支原体肺炎中的应用观察

目的分析金花清感颗粒联合布地奈德、沙丁胺醇、异丙托溴铵三联雾化吸入治疗儿童肺炎支原体肺炎(MPP)的效果。方法前瞻性选取2022年2月—2023年2月常熟市第二人民医院收治的114例MPP患儿为研究对象,按照随机数字表法分为对照组、研究组,每组57例。对照组给予布地奈德、沙丁胺醇、异丙托溴铵三联雾化吸入治疗,研究组在对照组基础上给予金花清感颗粒,连续治疗10 d评估效果。比较两组症状改善情况、肺功能、临床疗效、气道重塑指标、炎症因子、T淋巴细胞亚群及药物不良反应发生情况。结果研究组发热消退时间、咳嗽消失时间、肺部啰音消失时间均短于对照组(P<0.05)。研究组治疗前后的达峰时间比、达峰容积比、呼吸频率的差值均高于对照组(P<0.05)。研究组总有效率高于对照组(P<0.05)。研究组治疗前后的气道壁厚度/外径比值、气道面积/总横截面积比值的差值均高于对照组(P<0.05)。研究组治疗前后的超敏C反应蛋白、白细胞介素-6、肿瘤坏死因子-α的差值均高于对照组(P<0.05)。研究组治疗前后的CD4^(+)、CD8^(+)、CD4^(+)/CD8^(+)的差值均高于对照组(P<0.05)。两组总不良反应发生率比较,差异无统计学意义(P>0.05)。结论金花清感颗粒联合布地奈德、沙丁胺醇、异丙托溴铵三联雾化吸入治疗儿童MPP疗效显著,可改善肺功能,抗气道重塑,抑制炎症反应,改善T淋巴细胞亚群,且安全性良好。

糖肾康调控NLRP3-Caspase-1-IL-18/IL-1β轴改善糖尿病肾病足细胞焦亡的机制研究

目的基于肾虚湿瘀理论探讨以益肾活血清泄为原则组方制备的糖肾康颗粒通过调控NOD样受体蛋白3(NOD-like receptor protein 3,NLRP3)-胱天蛋白酶-1(Caspase-1)-白介素(interleukin,IL)-18/IL-1β轴改善糖尿病肾病足细胞损伤的机制。方法动物实验:将18只db/db小鼠随机分为模型组、缬沙坦组、糖肾康组,每组6只。6只db/m小鼠作为对照组。糖肾康组给予1.17 g/(kg·d)灌胃,缬沙坦组予缬沙坦悬液10.4 mg/(kg·d)灌胃,对照组及模型组予等量生理盐水灌胃。疗程8周,观察db/db小鼠一般情况及糖代谢情况,通过肾脏组织苏木素—伊红、马松与过碘酸雪夫氏染色及透射电镜观察肾脏病理变化,通过生化分析或酶联免疫吸附测定法检测肾损伤指标包括尿白蛋白/肌酐、血肌酐、尿素氮、胱抑素C、β2微球蛋白、N-乙酰-β-D-氨基葡萄糖苷酶,Western blot及免疫荧光检测db/db小鼠肾脏肾病蛋白Nephrin表达情况,以及细胞焦亡炎性因子NLRP3、Caspase-1、IL-18及IL-1β的表达变化。细胞实验:采用30 mmol/L葡萄糖培养基诱导小鼠足细胞48小时建立足细胞损伤模型。制备糖肾康大鼠含药血清,采用噻唑蓝法筛选出糖肾康含药血清的最佳作用浓度。采用流式细胞仪检测糖肾康对高糖诱导的足细胞凋亡率的影响,收集各组细胞培养上清液,检测乳酸脱氢酶水平,Western blot法观察NLRP3、Caspase-1,酶联免疫吸附测定法检测IL-18及IL-1β的表达变化。结果糖肾康可改善db/db小鼠糖代谢,与模型组比较,糖肾康组及缬沙坦组的尿白蛋白/肌酐均明显降低(P<0.01),亦改善了N-乙酰-β-D-氨基葡萄糖苷酶(P<0.01)、β2微球蛋白(P<0.05,P<0.01)、血肌酐(P<0.01)、尿素氮(P<0.01)及胱抑素C(P<0.01)等肾损伤指标;糖肾康组Nephrin的表达明显增加(P<0.01),且优于缬沙坦组(P<0.05)。糖肾康可明显降低小鼠肾组织细胞焦亡炎性因子NLRP3(P<0.05)、Caspase-1(P<0.01)、IL-18(P<0.01)及IL-1β(P<0.01)的表达。糖肾康组可改善肾小球基底膜增厚,改善足细胞融合和内皮细胞窗孔结构和系膜基质堆积等糖尿病肾脏病理改变。噻唑蓝法筛选出糖肾康含药血清的最佳浓度为3.84 g/kg,此浓度下足细胞活力最强,凋亡率最低。糖肾康组可显著降低高糖诱导的足细胞LDH释放率(P<0.01),降低NLRP3(P<0.05)、Caspase-1(P<0.05)、IL-18(P<0.01)及IL-1β(P<0.01)的表达。结论糖肾康颗粒可能通过调控NLRP3-Caspase-1-IL-18/IL-1β通路抑制糖尿病肾病足细胞焦亡,减少炎症反应,保护足细胞,减轻糖尿病肾病肾损伤。

中成药治疗慢性阻塞性肺疾病稳定期的网状Meta分析

目的:采用网状Meta分析评价口服中成药治疗慢性阻塞性肺疾病(COPD)稳定期的有效性及安全性。方法:计算机检索国家知识基础设施数据库(CNKI)、中国学术期刊数据库(CSPD)、中文科技期刊数据库(CCD)、中国生物医学文献数据库(CBM)、PubMed、Embase、Web of Science、Cochrane library数据库中不同中成药治疗COPD稳定期的临床随机对照试验(RCT),检索时限为建库至2022年05月11日。采用R 4.1.1、Stata 16.0、Review manager 5.3软件进行数据分析。结果:共纳入70项RCT,包括6652例患者,涉及5种中成药。依据累计概率值排序,网状Meta分析结果显示,改善COPD自我评估测试(CAT)问卷评分方面,百令胶囊+常规治疗(>常规治疗;改善第一秒用力呼气容积占预计值百分比(FEV_(1)%)方面,百令胶囊+常规治疗>金水宝胶囊+常规治疗>补肺活血胶囊+常规治疗>玉屏风颗粒+常规治疗>常规治疗);改善六分钟步行距离(6MWD)方面,金水宝胶囊+常规治疗>补肺活血胶囊+常规治疗>百令胶囊+常规治疗>常规治疗;降低二氧化碳分压方面(PaCO_(2)),补肺活血胶囊+常规治疗>金水宝胶囊+常规治疗>百令胶囊+常规治疗>常规治疗;降低年急性发作次数方面,百令胶囊+常规治疗>常规治疗。5种中成药均未产生严重不良反应。结论:5种中成药治疗稳定期COPD均能提高临床综合疗效,但各有侧重,其中百令胶囊组综合疗效较好。

活血消异颗粒抑制子宫内膜异位症保守术后复发的双中心、随机对照研究

目的评价活血消异颗粒抑制子宫内膜异位症(EMs)保守术后复发的有效性与安全性,为中药抑制EMs术后复发提供依据。方法选取EMs术后气滞血瘀型患者72例作为研究对象,随机分为中药组和西药组,各36例,中药组予活血消异颗粒治疗,西药组予促性腺激素释放激素激动剂(GnRH-α)治疗。观察2组术后复发率、中医证候积分、糖类抗原125(CA125)、痛经评分、盆腔痛评分、妊娠率、血清性激素[雌二醇(E_(2))、促黄体生成素(LH)、卵泡刺激素(FSH)]水平及安全性指标变化情况。结果术后9个月、术后12个月,2组患者中医证候总积分均显著降低(P<0.05),中药组优于西药组(P<0.05);术后12个月,中药组中医临床总有效率优于西药组;术后2组患者血清CA125水平、痛经评分及盆腔痛严重程度均显著降低(P<0.05,P<0.01);治疗期间中药组总不良反应发生率低于西药组(P<0.01);中药组复发率稍低于西药组,但无统计学差异(P>0.05)。结论活血消异颗粒可明显改善EMs术后气滞血瘀证患者的中医证候,安全性较好,且与GnRH-α在预防EMs术后复发方面疗效相当,值得临床推广应用。

益气活血类方对心肌梗死恢复期病人心功能、中医证候的影响

目的:观察益气活血类方对心肌梗死恢复期病人心功能、中医证候的影响。方法:选取2021年5月—2022年12月首都医科大学附属北京中医医院及北京中医医院平谷医院门诊及住院治疗的心肌梗死恢复期病人144例作为研究对象,采用随机数字法将病人分为补阳还五颗粒组(31例)、脑心通胶囊组(49例)、养阴通脑颗粒组(31例)、对照组(33例)。补阳还五颗粒组给予标准化治疗+补阳还五颗粒+胶囊模拟剂,脑心通胶囊组给予标准化治疗+脑心通胶囊+颗粒模拟剂,养阴通脑颗粒组给予标准化治疗+养阴通脑颗粒+胶囊模拟剂,对照组给予标准化治疗+颗粒模拟剂+胶囊模拟剂。连续治疗3个月。比较各组左室射血分数(LVEF)、左心室舒张末期内径(LVEDD)、左心室收缩末期内径(LVESD)、N末端B型利钠肽原(NT-proBNP)水平、NT-proBNP变化率、中医证候积分及中医证候疗效。结果:治疗后,补阳还五颗粒组、脑心通胶囊组及养阴通脑颗粒组LVEF水平较治疗前升高,且优于对照组(P<0.05)。补阳还五颗粒组、脑心通胶囊组及养阴通脑颗粒组治疗前后LVEDD、LVESD水平比较差异无统计学意义(P>0.05)。治疗后,补阳还五颗粒组、脑心通胶囊组NT-proBNP水平均较治疗前下降(P<0.05)。以NT-proBNP下降30%为标准,补阳还五颗粒组NT-proBNP下降≥30%的病人较其他3组多(P<0.05)。治疗后,脑心通胶囊组及养阴通脑颗粒组胸痛积分均优于对照组(P<0.05);脑心通胶囊组出汗积分较治疗前下降(P<0.05);脑心通胶囊组面色紫暗积分及总分均优于对照组(P<0.05)。补阳还五颗粒能缓解病人神疲乏力的症状,脑心通胶囊能缓解病人胸闷、气短的症状且能更加全面地减轻病人临床症状。结论:补阳还五颗粒、脑心通胶囊、养阴通脑颗粒能够提高心肌梗死恢复期病人心功能水平,改善临床症状。

清肝利湿颗粒联合恩替卡韦治疗肝胆湿热型乙肝肝硬化的临床疗效观察

目的探讨清肝利湿颗粒联合恩替卡韦治疗肝胆湿热型乙型肝炎(以下简称“乙肝”)肝硬化的临床效果。方法选取2021年1月至2023年4月肇庆市中医院收治的100例肝胆湿热型乙肝肝硬化患者作为研究对象,按照随机抽签的方式分为对照组(50例)和研究组(50例)。对照组采用恩替卡韦治疗,研究组采用恩替卡韦与清肝利湿颗粒联合治疗。比较两组患者临床疗效、生化指标(丙氨酸转氨酶、天冬氨酸转氨酶、总胆红素)、影像学特征、乙肝病毒DNA转阴率以及不良反应发生情况。结果研究组治疗总有效率高于对照组,差异有统计学意义(P<0.05)。治疗后,研究组患者丙氨酸转氨酶、天冬氨酸转氨酶、总胆红素水平均低于对照组,差异有统计学意义(P<0.05)。治疗后,研究组患者门静脉内径、门静脉最大流速、脾静脉内径、脾静脉最大流速均低于对照组,差异有统计学意义(P<0.05)。研究组乙肝病毒DNA转阴率高于对照组,差异有统计学意义(P<0.05)。研究组发生4例不良反应,对照组发生6例不良反应,两组患者不良反应总发生率比较,差异无统计学意义(P>0.05)。结论清肝利湿颗粒联合恩替卡韦治疗肝胆湿热型乙肝肝硬化的效果显著,可有效抑制乙型肝炎病毒复制,改善患者肝功能,且安全性较高。

金花清感颗粒治疗新型冠状病毒感染的理论探析

金花清感颗粒研发于甲型H1N1流感病毒疫情盛行时期,其组方以经典名方银翘散和麻杏石甘汤为主体传承创新而成,主治病机为风温初起、卫气同病,其中知母为组方的点睛之笔,起到坚阴护卫之效。金花清感颗粒在新型冠状病毒感染疫情防治中均展现出很高的临床疗效,其在新型冠状病毒感染防治中主要针对的证型为:燥伏太阴、伏燥兼湿、卫气同病,进一步强调了伏燥病机在新型冠状病毒感染发病中的重要性,为今后的防治提供了新思路。

益气活血固肾颗粒中5种有效成分在大鼠体内的药动学及组织分布研究

目的探究益气活血固肾颗粒中巴马汀、药根碱、小檗碱、表小檗碱、芒柄花素在大鼠体内的药动学特征及组织分布规律,为进一步阐明该方的药效物质基础提供依据。方法建立HPLC-MS/MS法测定大鼠口服给药4.75 g/kg益气活血固肾颗粒后血浆及各组织中的药物浓度,色谱柱选择Kromasil C_(18)柱(3.5μm,2.1 mm×100 mm),流动相为乙腈-0.1%甲酸水溶液,流速为0.3 mL/min,进样量5μL,采用电喷雾电离源(ESI)分别在正、负离子多反应监测模式(MRM)下进行扫描,选择WinNonlin 3.3软件计算主要药动学参数。结果所建立的分析方法符合生物样品的检测要求,给予复方灌胃药液后,5种成分均可快速吸收入血,在30 min内血药浓度达到高峰,且药物在体内半衰期较短,消除速率快。各成分的达峰浓度Cmax、药时曲线下面积AUC_(0-t)分别为4.87~233.12μg/L、16.11~493.13 h/(μg·L)。组织分布结果显示,5种成分在体内分布迅速且广泛,具有不均衡的特征,在胃和小肠中分布较多,其次是肝、肾、肺等组织,而在脾脏、心脏中分布较少。结论该方法稳定、可靠,为复方益气活血固肾颗粒临床应用提供了参考。

柴黄清胰活血颗粒通过抑制NLRP3炎症小体活化减轻重症急性胰腺炎模型大鼠炎症损伤的机制研究

目的通过观察柴黄清胰活血颗粒对NLRP3炎症小体活化的调控作用,探讨其对重症急性胰腺炎(severe acute pancreatitis,SAP)模型大鼠胰腺组织的作用及机制。方法将64只SD大鼠随机分为假手术组、重症急性胰腺炎模型组、柴黄清胰活血颗粒组及MCC950(NLRP3抑制剂)组,各组再分为12 h和24 h两个亚组,每组8只。重症急性胰腺炎模型组、柴黄清胰活血颗粒组及MCC950组给予3.5%的牛磺胆酸钠(2 mL·kg^(-1))逆行胰胆管注射构建重症急性胰腺炎模型。MCC950组于模型制备完成后立即腹腔注射MCC950(1 mg·mL-1)。麻醉苏醒后,柴黄清胰活血颗粒组灌胃柴黄清胰活血颗粒溶液(0.35 g·mL-1),6 h一次。造模后12 h和24 h收集腹水、腹主动脉血及胰腺组织。采用全自动生化分析仪检测血清淀粉酶及脂肪酶活性;HE染色观察胰腺损伤情况;ELISA法检测血清IL-18、IL-1β的变化,IHC、qRT-PCR及Western Blot法检测NLRP3炎症小体活化相关基因及蛋白的表达。结果与重症急性胰腺炎组比较,柴黄清胰活血颗粒组及MCC950组的胰腺组织病理损伤明显减轻,病理评分明显降低(P<0.05),血清脂肪酶、淀粉酶、白细胞介素(IL)-18、IL-1β水平明显降低(P<0.05);与重症急性胰腺炎组比较,经柴黄清胰活血颗粒治疗或腹腔注射NLRP3抑制剂后,胰腺组织NLRP3、ASC、Caspase-1免疫组化阳性表达量及NLRP3、ASC、Caspase-1 mRNA表达水平和NLRP3、ASC、Pro-Caspase-1、Caspase-1蛋白表达水平明显降低(P<0.05)。结论柴黄清胰活血颗粒可通过抑制NLRP3炎症小体活化,降低胰腺组织NLRP3、ASC、Caspase-1 mRNA及蛋白的表达,降低下游炎症因子IL-18、IL-1β的释放,从而减轻重症急性胰腺炎模型大鼠胰腺炎症损伤。

基于网络药理学分析金花清感颗粒抗炎机制及验证

目的利用网络药理学和体外细胞试验,研究金花清感颗粒的抗炎活性及机制,为其临床应用提供理论依据。方法利用TCMSP、Pubchem数据库和相关文献筛选出金花清感颗粒的活性成分及靶标;通过NCBI、GenCards数据库筛选流行性感冒、头痛、发热、咳嗽的关联靶点,并与上述成分-靶点进行比对分析,筛选出核心靶点;采用Cytoscape 3.7.2软件生成药物-关键靶点网络图,最后利用Metascape数据库对核心靶点进行GO、KEGG富集分析。体外细胞试验采用脂多糖(LPS)诱导小鼠单核巨噬细胞RAW264.7炎症细胞模型,采用Griess法检测金花清感颗粒提取物对细胞一氧化氮(NO)分泌的影响;通过实时荧光定量PCR法(qRTPCR)和Western blotting分别测定金花清感颗粒提取物对RAW264.7细胞中关键靶标的RNA和蛋白水平的影响。结果数据库筛选后共获得金花清感颗粒活性成分278个,潜在作用靶点1316个,与疾病相关靶点1065个,主要涉及丝裂原活化蛋白激酶(MAPK)信号通路、核因子κB(NF-κB)信号通路及神经活性配体-受体相互作用信号通路等。体外细胞试验结果显示,与LPS组相比,200,400,800μg/ml的金花清感颗粒水、醇提物均显著降低LPS诱导的RAW264.7细胞中NO的分泌量(P<0.01);qRT-PCR检测发现金花清感颗粒提取物能抑制LPS诱导的RAW264.7细胞的诱导型一氧化氮合酶(iNOS),环氧合酶2(COX-2),白介素6(IL-6)mRNA水平(P<0.01);Western blotting检测发现金花清感颗粒提取物能抑制LPS诱导的RAW264.7细胞的iNOS蛋白的表达及NF-κB、MAPK信号通路中关键蛋白的活化。结论基于网络药理学研究显示金花清感颗粒具有多靶点、多通路协同作用发挥药效的特点,通过体外细胞试验进一步验证显示金花清感颗粒通过抑制炎症细胞模型iNOS表达和NF-κB与MAPK信号通路的激活干预炎症细胞因子释放,发挥抗炎活性,该结果为深入研究金花清感颗粒作用机制及临床应用提供理论依据。

清肝化湿活血汤加减联合茵栀黄颗粒治疗酒精性肝病临床观察

目的探讨清肝化湿活血汤加减联合茵栀黄颗粒治疗酒精性肝病的临床疗效。方法选取医院2021年3月至2023年3月收治的酒精性肝病患者200例,按随机数字表法分为观察组和对照组,各100例。两组患者均予酒精性肝病的基础治疗,对照组患者加用茵栀黄颗粒,观察组患者在对照组治疗基础上加用清肝化湿活血汤加减。两组患者均治疗12周。结果观察组总有效率为91.00%,显著高于对照组的79.00%(P<0.05)。治疗后,观察组患者肋痛胀满、痞块结坚、乏力纳呆、舌红苔黄的中医证候积分均显著低于对照组(P<0.05);观察组患者的丙氨酸氨基转移酶、天门冬氨酸氨基转移酶、碱性磷酸酶、总胆红素水平均显著低于对照组(P<0.05);观察组患者的肝门静脉血流动力学指标最大流速、平均流速、搏动指数均显著高于对照组(P<0.05),阻力指数显著低于对照组(P<0.05);观察组患者的血清炎性因子白细胞介素17A、前列腺素E_(2)、CXC趋化因子配体13水平均显著低于对照组(P<0.05);观察组患者的欧洲研究与治疗组织生活满意度测定量表(EORTCQLQ-C30)躯体功能、角色功能、社会功能、总体健康评分均显著高于对照组(P<0.05)。治疗期间,观察组和对照组不良反应发生率相当(12.00%比8.00%,P>0.05)。结论清肝化湿活血汤加减联合茵栀黄颗粒治疗酒精性肝病的临床疗效良好,可改善患者的中医证候积分和肝功能、肝门静脉血流动力学指标,降低炎性因子水平,且安全性良好。

柴黄清胰活血颗粒对重症急性胰腺炎模型大鼠肠道微生态变化的影响

目的探讨柴黄清胰活血颗粒调节重症急性胰腺炎(severe acute pancreatitis,SAP)模型大鼠肠道微生态变化和治疗重症急性胰腺炎的潜在机制。方法将48只大鼠随机分为假手术组、模型组、柴黄清胰活血颗粒组,每组16只;各组再随机分为12 h亚组和24 h亚组。以肠壁穿刺逆行胰胆管注射5%牛磺胆酸钠复制重症急性胰腺炎大鼠模型。假手术组、模型组予以生理盐水灌胃,柴黄清胰活血颗粒组予以1.2 g·kg^(-1)柴黄清胰活血颗粒溶液灌胃,每间隔6 h灌胃1次。术后12 h、24 h,每组各取8只大鼠采集腹主动脉血、胰腺及回肠组织备测。观察大鼠腹腔内腹水、胰腺及回肠组织情况;生化法检测血清淀粉酶(amylase,AMY)、脂肪酶(lipase,LPS);HE染色观察胰腺及回肠组织病理变化;Western Blot法检测回肠组织中Claudin-1蛋白;16S rDNA高通量测序检测回盲部内容物肠道菌群变化情况。结果与相应时点的假手术组比较,模型组大鼠胰腺广泛水肿坏死,血清淀粉酶、脂肪酶明显升高,胰腺组织、回肠组织病理评分升高,回肠组织Claudin-1蛋白表达量明显下降(均P<0.05);微生物群落的丰度差异性变大,群落组成的均匀度变低,菌群的丰富度无明显变化(P>0.05)而多样性减少(P<0.05),变形菌门为肠道优势菌,颤螺菌属、瘤胃球菌、双歧杆菌属及拟杆菌S24-7等菌相对丰度降低,志贺菌、Allobaculum菌等相对丰度增加;微生物群落的丰度差异性变小,群落组成的均匀度变高,菌群丰富度无明显变化(P>0.05),而多样性增加(P<0.05),以厚壁菌门、拟杆菌门为肠道优势菌,颤螺菌属、瘤胃球菌、双歧杆菌、拟杆菌S24-7等菌相对丰度增加,志贺菌、Allobaculum菌等相对丰度降低。经柴黄清胰活血颗粒干预后,回肠组织病理损伤改善,肠黏膜屏障Claudin-1蛋白表达量均较模型组增加(P<0.05);微生物群落的丰度差异性变小,群落组成的均匀度变高,较模型组部分有益菌增多,致病菌减少。结论柴黄清胰活血颗粒能减轻重症急性胰腺炎大鼠胰腺炎症损伤,其治疗作用可能与调节肠道微生态,改善肠黏膜屏障功能有关。

清肝活血方调控Caspase-4/Caspase-3/GSDME介导的细胞焦亡改善酒精性肝损伤

[目的]研究清肝活血方(QGHXR)通过调控半胱天冬酶(Caspase)-4/Caspase-3/GSDME细胞焦亡通路防治酒精性肝病(ALD)的作用机制。[方法]将18只C57BL/6小鼠随机分为对照组、模型组和QGHXR组复制ALD小鼠模型。检测血清肝功能、血脂水平;苏木精-伊红(HE)染色及油红O染色观察肝脏病理改变及肝脏脂质沉积状况;实时荧光定量逆转录聚合酶链反应(qRT-PCR)、蛋白免疫印迹(Western Blot)法及免疫荧光染色检测Caspase-4/Caspase-3/GSDME通路相关基因及蛋白表达情况。[结果]与空白对照组比较,模型组小鼠肝组织三酰甘油(TG)水平明显升高(P<0.01),血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)水平升高(P<0.05),血清白介素-1β(IL-1β)、白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)水平升高(P<0.05);肝脏组织出现病理改变及脂质沉积现象;肝组织IL-1B、IL-6、TNF-α、NLRP3 mRNA表达水平升高。与模型组比较,QGHXR组小鼠肝组织TG水平明显降低(P<0.01),血清ALT、AST、总胆红素(TBIL)、总胆汁酸(TBA)水平降低(P<0.05),血清TG、IL-6、TNF-α水平降低(P<0.05)以及血清高密度脂蛋白(HDL)水平升高(P<0.01);血清中血脂、肝脏病理改变及脂质沉积现象缓解;GsdmeN、Caspase-4、Caspase-3和C-Caspase-3基因及蛋白水平降低(P<0.05)。[结论]QGHXR可通过调节Caspase-4/Caspase-3/GSDME介导的细胞焦亡通路改善ALD小鼠肝损伤和脂质沉积。