Integrated serum pharmacochemistry and investigation of the anti-influenza A virus pneumonia effect of Qingjin Huatan decoction.

ETHNOPHARMACOLOGICAL RELEVANCE: Qingjin Huatan Decoction (QJHTT) consists of 11 herbal medicines: Scutellaria baicalensis Georgi, Gardenia jasminoides J. Ellis, Platycodon grandiflorus (Jacq.) A. DC., Ophiopogon japonicus (Thunb.) Ker Gawl., Morus alba L., Fritillaria thunbergii Miq., Anemarrhena asphodeloides Bunge, Trichosanthes kirilowii Maxim., Citrus reticulata Blanco, Poria cocos (Schw.) Wolf, and Glycyrrhiza uralensis Fisch. As a traditional Chinese medicinal formula, QJHTT has been used for more than 400 years in China. It has shown promising results in treating influenza A virus (IAV) pneumonia. AIM OF THE STUDY: To elusive the specific pharmacological constituents and mechanisms underlying its anti-IAV pneumonia effects. MATERIALS AND METHODS: The components in QJHTT were analyzed through the use of a serum pharmacology-based ultra high-performance liquid chromatography Q- Exactive Orbitrap mass spectrometry (UHPLC-Q Exactive Orbitrap-MS) method. Simultaneously, the dynamic changes in IAV-infected mouse lung viral load, lung index, and expression of lung inflammation factors were monitored by qRT-PCR. RESULTS: We successfully identified 152 chemical components within QJHTT, along with 59 absorbed chemical prototype constituents found in the serum of mice treated with QJHTT. 43.45% of these chemical components and 43.10% of the prototype constituents were derived from the monarch drugs, namely Huangqin and Zhizi, aligning perfectly with traditional Chinese medicine theory. Notably, our analysis led to the discovery of 14 compounds within QJHTT for the first time, three of which were absorbed into the bloodstream. Simultaneously, we observed that QJHTT not only reduced the viral load but also modulated the expression of inflammation factors in the lung tissue including TNF-α, IL-1ß, IL-4, IL-6, IFN-γ, and IL17A. A time-effect analysis further revealed that QJHTT intervention effectively suppressed the peak of inflammatory responses, demonstrating a robust anti-IAV pneumonia effect. CONCLUSIONS: We comprehensively analyzed the pharmacological material basis of QJHTT by a highly sensitive and high-resolution UHPLC-Q Exactive Orbitrap-MS method, and demonstrated its efficacy in combating IAV pneumonia by reducing lung viral load and inflammatory factors. This study has significant importance for elucidating the pharmacological basis and pharmacological mechanism of QJHTT in combating IAV pneumonia.

Qingjin Huatan decoction protects mice against influenza a virus pneumonia via the chemokine signaling pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: Qingjin Huatan Decoction (QJHTT) consists of 11 herbal medicines: Scutellaria baicalensis Georgi, Gardenia jasminoides J.Ellis, Platycodon grandiflorus (Jacq.) A.DC., Ophiopogon japonicus (Thunb.) Ker Gawl., Morus alba L., Fritillaria thunbergii Miq., Anemarrhena asphodeloides Bunge, Trichosanthes kirilowii Maxim., Citrus reticulata Blanco, Poria cocos (Schw.) Wolf, and Glycyrrhiza uralensis Fisch. As a traditional compound Chinese medicinal formula, QJHTT has been used for more than 400 years in China. Historically, it was used to treat respiratory diseases and had shown beneficial clinical results for diseases related to lung inflammation. AIM OF THE STUDY: To investigate the therapeutic effect of QJHTT on influenza A virus (IAV) pneumonia in mice and explore its possible mechanism of action. MATERIALS AND METHODS: The components in QJHTT were analyzed by UPLC-Q-TOF-MS and some antiviral active components reported in the literature were determined and quantified by HPLC. The protective effects of QJHTT were investigated using lethal and sublethal doses (2 LD50 or 0.8 LD50 viral suspension, separately) of H1N1-infected mice. Mortality and lung lesions in H1N1-infected mice were used to evaluate the efficacy of QJHTT. The potential mechanism of QJHTT in the treatment of viral pneumonia was determined at the gene level by RNA sequencing and validated by qRT-PCR. Following this, the changes in protein levels of JAK2/STAT3 were analyzed since it is a key downstream target of the chemokine signaling pathways. Preliminary elucidation of the mechanism of QJHTT to protect mice against IAV pneumonia through this pathway was conducted. RESULTS: In this study, 12 antiviral active constituents including baicalin, geniposide, and mangiferin were identified from QJHTT. In vivo treatment of QJHTT reduced the virus titers of lung tissue significantly and improved the survival rate, lung index, and pulmonary histopathological changes; additionally, a reduction in the serum levels of TNF-α, IL-1ß, IL-6, and IFN-γ inflammatory factors in H1N1-infected mice was observed. RNA-seq analysis and qRT-PCR showed that QJHTT primarily reversed the activities CCL2, CCL7, CCR1, and other chemokines and their reception-related genes, suggesting that QJHTT may produce disease-resistant pneumonia by inhibiting the downstream JAK2/STAT3 pathway. Western blot analysis confirmed that QJHTT effectively reduced the protein levels of JAK2, STAT3, and related phosphorylated products in the lung tissue of H1N1-infected mice. CONCLUSIONS: Our results indicated that QJHTT alleviated IAV pneumonia in mice by regulating related chemokines and their receptor-related genes in lung tissue, thereby inhibiting JAK2/STAT3 pathway. This could pave way for the design of novel therapeutic strategies to treat viral pneumonia.

[Study on critical quality attributes of Qingjin Huatan Decoction based on serum pharmacochemistry].

Qingjin Huatan Decoction is a classic prescription with the effects of clearing heat, moistening lung, resolving phlegm, and relieving cough. In order to explore the critical quality attributes of Qingjin Huatan Decoction, we identified the blood components of Qingjin Huatan Decoction by ultra-performance liquid chromatography quadrupole time of flight mass spectrometry(UPLC-Q-TOF-MS) under the following conditions, chromatographic column: Acquity UPLC BEH C_(18) column(2.1 mm×100 mm, 1.7 µm); mobile phase: 0.1% formic acid acetonitrile(A)-0.1% formic acid in water(B); gradient elution; flow rate: 0.2 mL·min~(-1); column temperature: 30 ℃; injection volume: 5 µL. The electrospray ionization(ESI) source was used to collect data in both positive and negative ion modes under the following conditions, capillary voltage: 3 kV for the positive ion mode and 2 kV for the negative ion mode; ion source temperature: 110 ℃; cone voltage: 30 V; cone gas flow rate: 50 L·h~(-1); nitrogen degassing temperature: 350 ℃; degassing volume flow rate: 800 L·h~(-1); scanning range: m/z 50-2 000. In this experiment, a total of 66 related components of Qingjin Huatan Decoction were identified, including 22 prototype components and 44 metabolites. The results of this study preliminarily revealed the pharmacodynamic material basis of Qingjin Huatan Decoction in vivo, which has provided an experimental basis for the determination of quality markers of Qingjin Huatan Decoction and the development of new drugs.

Study on critical quality attributes of Qingjin Huatan Decoction based on serum pharmacochemistry / 中国中药杂志

Qingjin Huatan Decoction is a classic prescription with the effects of clearing heat, moistening lung, resolving phlegm, and relieving cough. In order to explore the critical quality attributes of Qingjin Huatan Decoction, we identified the blood components of Qingjin Huatan Decoction by ultra-performance liquid chromatography quadrupole time of flight mass spectrometry(UPLC-Q-TOF-MS) under the following conditions, chromatographic column: Acquity UPLC BEH C_(18) column(2.1 mm×100 mm, 1.7 μm); mobile phase: 0.1% formic acid acetonitrile(A)-0.1% formic acid in water(B); gradient elution; flow rate: 0.2 mL·min~(-1); column temperature: 30 ℃; injection volume: 5 μL. The electrospray ionization(ESI) source was used to collect data in both positive and negative ion modes under the following conditions, capillary voltage: 3 kV for the positive ion mode and 2 kV for the negative ion mode; ion source temperature: 110 ℃; cone voltage: 30 V; cone gas flow rate: 50 L·h~(-1); nitrogen degassing temperature: 350 ℃; degassing volume flow rate: 800 L·h~(-1); scanning range: m/z 50-2 000. In this experiment, a total of 66 related components of Qingjin Huatan Decoction were identified, including 22 prototype components and 44 metabolites. The results of this study preliminarily revealed the pharmacodynamic material basis of Qingjin Huatan Decoction in vivo, which has provided an experimental basis for the determination of quality markers of Qingjin Huatan Decoction and the development of new drugs.

[Meta-analysis and trial sequential analysis of Qingjin Huatan Decoction for treating community-acquired pneumonia in elderly].

To systematically evaluate the clinical efficacy and safety of modified Qingjin Huatan Decoction in the treatment of community-acquired pneumonia in the elderly, and provide evidence-based reference for the clinical application of this prescription. Randomized controlled trials of Qingjin Huatan Decoction in the treatment of community-acquired pneumonia in the elderly were collected by searching PubMed, EMbase, Cochrane Library, CNKI, China Biomedical Literature database, VIP database and WanFang database. Outcome indicators included clinical effective rate, inflammation index, symptom improvement time, chest radiograph improvement time, hospitalization time and adverse reactions. RevMan 5.3 and Stata/IC 15.1 software were used for Meta-analysis; TSA 0.9.5.10 Beta software was used for trial sequential analysis, and GRADE profiler 3.6 was used for grade evidence quality evaluation. Thirteen studies were included finally, including 1 058 patients, 536 patients in the experimental group and 522 patients in the control group.Meta-analysis showed that, the clinical effective rate of the experimental group was significantly higher than that of the control group(RR=1.16, 95%CI[1.10, 1.21], P<0.000 01); fever time(MD=-1.32, 95%CI[-1.93,-0.71], P<0.000 1), cough time(MD=-1.95, 95%CI[-2.69,-1.21), P<0.000 01), time to rale disappearance(MD=-1.55, 95%CI[-2.37,-0.73], P=0.000 2), time to chest radiograph improvement(MD=-1.72, 95%CI[-2.98,-0.46], P=0.007), and hospitalization time(MD=-3.16, 95%CI[-4.58,-1.74], P<0.000 01) in the experimental group were significantly shorter than those in the control group. The improvement in CRP(WMD=-3.44,95%CI[-4.50,-2.38],P<0.001), WBC(WMD=-2.04,95%CI[-3.31,-0.78],P<0.01), IL-6(WMD=-4.27,95%CI[-4.62,-3.92],P<0.001), and TNF-α(WMD=-0.47,95%CI[-0.55,-0.39], P<0.001) of the experimental group was significantly better than that of the control group. There was no significant difference in PCT improvement between the two groups(WMD=-0.63, 95%CI[-1.65, 0.40], P=0.23). No serious adverse reactions occurred in both groups. Sequential analysis of the trial showed that in the studies with cumulative inclusion of clinical effective rate, the data passed the traditional threshold and TSA threshold, further confirming its clinical efficacy. GRADE evaluation showed that the evidence level was low to extremely low. Western medicine treatment combined with Qingjin Huatan Decoction may improve clinical efficiency in the treatment of community-acquired pneumonia in the elderly, improve clinical symptoms faster, reduce CRP, WBC and other inflammatory indicators, and shorten hospital stay. The level of evidence obtained in this study is low, which needs to be further verified by high-quality multi-center, randomized controlled trials.

Meta-analysis and trial sequential analysis of Qingjin Huatan Decoction for treating community-acquired pneumonia in elderly / 中国中药杂志

To systematically evaluate the clinical efficacy and safety of modified Qingjin Huatan Decoction in the treatment of community-acquired pneumonia in the elderly, and provide evidence-based reference for the clinical application of this prescription. Randomized controlled trials of Qingjin Huatan Decoction in the treatment of community-acquired pneumonia in the elderly were collected by searching PubMed, EMbase, Cochrane Library, CNKI, China Biomedical Literature database, VIP database and WanFang database. Outcome indicators included clinical effective rate, inflammation index, symptom improvement time, chest radiograph improvement time, hospitalization time and adverse reactions. RevMan 5.3 and Stata/IC 15.1 software were used for Meta-analysis; TSA 0.9.5.10 Beta software was used for trial sequential analysis, and GRADE profiler 3.6 was used for grade evidence quality evaluation. Thirteen studies were included finally, including 1 058 patients, 536 patients in the experimental group and 522 patients in the control group.Meta-analysis showed that, the clinical effective rate of the experimental group was significantly higher than that of the control group(RR=1.16, 95%CI[1.10, 1.21], P<0.000 01); fever time(MD=-1.32, 95%CI[-1.93,-0.71], P<0.000 1), cough time(MD=-1.95, 95%CI[-2.69,-1.21), P<0.000 01), time to rale disappearance(MD=-1.55, 95%CI[-2.37,-0.73], P=0.000 2), time to chest radiograph improvement(MD=-1.72, 95%CI[-2.98,-0.46], P=0.007), and hospitalization time(MD=-3.16, 95%CI[-4.58,-1.74], P<0.000 01) in the experimental group were significantly shorter than those in the control group. The improvement in CRP(WMD=-3.44,95%CI[-4.50,-2.38],P<0.001), WBC(WMD=-2.04,95%CI[-3.31,-0.78],P<0.01), IL-6(WMD=-4.27,95%CI[-4.62,-3.92],P<0.001), and TNF-α(WMD=-0.47,95%CI[-0.55,-0.39], P<0.001) of the experimental group was significantly better than that of the control group. There was no significant difference in PCT improvement between the two groups(WMD=-0.63, 95%CI[-1.65, 0.40], P=0.23). No serious adverse reactions occurred in both groups. Sequential analysis of the trial showed that in the studies with cumulative inclusion of clinical effective rate, the data passed the traditional threshold and TSA threshold, further confirming its clinical efficacy. GRADE evaluation showed that the evidence level was low to extremely low. Western medicine treatment combined with Qingjin Huatan Decoction may improve clinical efficiency in the treatment of community-acquired pneumonia in the elderly, improve clinical symptoms faster, reduce CRP, WBC and other inflammatory indicators, and shorten hospital stay. The level of evidence obtained in this study is low, which needs to be further verified by high-quality multi-center, randomized controlled trials.

Clinical Observation of Qingjin Huatan Decoction for Patients with AECOPD Phlegm-heat Obstructing Lung / 中国药房

OBJECTIVE:To observe the clinical efficacy and safety of Qingjin huatan decoction for patients with AECOPD phlegm-heat obstructing lung. METHODS:120 patients with AECOPD phlegm-heat obstructing lung were divided into observation group and control group according to random number table,with 60 cases in each group. Control group received routine therapy as low-flow oxygen therapy,anti-infective,bronchiectasis,expectorant,etc.;observation group additionally received Qingjin huatan decoction(decocted by 400 ml pure water),tid,on the basis of control group. The levels of IL-8,IL-17,IL-23 and TNF-α were observed in 2 groups before and 7 d after treatment. Clinical efficacies and the occurrence of ADR were compared between 2 groups. RESULTS:Before treatment,there was no statistical significance in the levels of IL-8,IL-17,IL-23 and TNF-α between 2 groups (P>0.05). After treatment,the levels of IL-8,IL-17,IL-23 and TNF-α in observation group were significantly decreased and lower than in control group,with statistical significance (P0.05). Excellent rate of observation group was 95.00%,which was significantly higher than 78.95% of control group,with statistical significance(P0.05). CONCLUSIONS:Qingjin huatan decoction shows signifi-cant therapeutic efficacy in the treatment of AECOPD phlegm-heat obstructing lung,and can improve inflammatory factors with good safety.

Effects of Qingjin Huatan Decoction on EGFR/MAPK Signaling Pathway of Airway Mucus Hypersecretion Rats with Chronic Obstructive Pulmonary Disease / 中国中医药信息杂志

Objective To observe the effects of Qingjin Huatan Decoction on EGFR/MAPK signaling pathway of airway mucus hypersecretion rats with chronic obstructive pulmonary disease (COPD). Methods Intratracheal instillation of LPS combined with smudging method was used to establish COPD airway mucus hypersecretion rat models. Experimental rats were randomly divided into blank group, model group, Qingjin Huatan Decoction group and clarithromycin group. The blank group was normally fed, while the other three groups were given NS, Qingjin Huatan Decoction, and clarithromycin respectively for gavage, once a day for 30 days. All rats were killed on the 31st day, and pathological changes of lung tissue and mucous glands hyperplasia were observed by HE staining method. The gene expressions of EGFR and MUC5AC in lung tissue were detected by RT-PCR method. The protein expressions of P-EGFR, P-ERK, P-JNK, P-p38 and MUC5AC in pulmonary tissue and airway epithelium were detected by immunohistochemical method. Results Compared with the blank group, mucous glands hyperplasia on airway epithelium, protein expressions of P-EGFR, P-ERK, P-JNK, P-p38 and MUC5AC on airway epithelium significantly increased in the model group (P<0.01); gene expression of MUC5AC of lung tissue increased (P<0.05). Compared with the model group, mucous glands hyperplasia on airway epithelium, P-p38, P-ERK and MUC5AC protein expression on airway epithelium in Qingjin Huatan Decoction group significantly decreased (P<0.05, P<0.01); the protein expression of P-JNK increased significantly (P<0.01). EGFR and MUC5AC mRNA in lung tissue in Qingjin Huatan Decoction group decreased significantly (P<0.01). Conclusion Qingjin Huatan Decoction can reduce airway mucus hepersecrection of COPD by inhibiting ERK and p38 signal pathway on EGFR downstream.

Clinical study on pulmonary complications after thoracotomy in elderly patients treated with atomization inhalation ofQingjin-Huatandecoction / 国际中医中药杂志

Objective The study was designed to investigate the effect of atomization inhalation of Qingjin-Huatandecoction on prevention of elderly patients’ postoperative pulmonary complications after thoracotomy.MethodsA total of 60 elderly patients with thoracotomy were randomized into 2 groups. Both groups of patients were treated with oxygen uptake, anti-infection, antispasmodic, analgesic, and nutritional support. Based on the conventional treatment, the control group was added with atomization inhalation of mucosolvan and dexamethasone, while the treatment group with atomization inhalation ofQingjin-Huatan decoction. The volume of phlegm, serum CRP, TNF-α level and absolute neutrophil count (ANC), arterial blood gasanalysis, and records of the pulmonary complications were meausred.Results The pH value at Day 3 (7.39 ± 0.06vs. 7.36 ± 0.05,t=2.120), the PaO2at Day 3 (86.51 ± 3.96 mmHgvs. 84.07 ± 3.51 mmHg,t=2.520) and Day 5 (92.27 ± 2.94 mmHgvs. 90.40 ± 3.08 mmHg,t=2.400) of treatment group were significantly higher than those of the control group; the PaCO2at Day 3 (44.57 ± 2.53 mmHgvs. 46.39 ± 3.35 mmHg,t=2.390) and  Day 5 (40.77 ± 2.48 mmHgvs. 42.42 ± 3.33 mmHg,t=2.130) of treatment group were significantly lower than those of the control group. The serum CRP at Day 3(8.26 ± 2.38 mg/Lvs. 9.76 ± 2.37 mg/L,t=2.440) and Day 5 (6.94 ± 1.76 mg/Lvs. 8.15 ± 2.38 mg/L,t=2.250), the ANC at Day 3 (8.54 ± 1.85)×109/Lvs. (9.58 ± 1.94)×109/L, t=2.140) and Day 5 (6.54 ± 1.85)×109/Lvs. (7.58 ± 1.95)×109/L,t=2.160), the serum TNF-a at Day 5 (7.54 ± 1.38 mg/Lvs. 8.43 ± 1.36 mg/L,t=2.540) of treatment group were significantly lower than those of the control group. The sputum volume at Day 3 (50.74 ± 18.46 ml/dvs. 59.92 ± 15.25 ml/d,t=2.099) and Day 5 (39.64 ± 18.21 ml/dvs. 48.99 ± 13.55 ml/d,t=2.256) of treatment group was lower than that of the control group. At Day 5, the pneumonia (3.33%vs. 20.00%,t=4.043) and the atelectasis (6.67%vs. 26.77%,t=4.320) of treatment group were significantly lower than those of the control group(P<0.05).ConclusionAtomization inhalation ofQingjin-Huatandecoction can improve pulmonary function of postoperative pulmonary complications of elderly patients after thoracotomy, take control of inflammation and the occurrence of postoperative pulmonary complications.

Effects of Qingjin Huatan Decoction on Expressions of NE, EGFR, and MUC5AC mRNA in ;Lung Tissue of COPD Rats / 中国中医药信息杂志

Objective To investigate the action mechanism of Qingjin Huatan Decoction on regulating COPD rats with mucus hypersecretion in airway. Methods Fifty 6-8 week old Wistar male rats were randomly divided into healthy group, model group, Qingjin Huatan Decoction group, clarithromycin group, 10 rats in each group. Except for healthy group, all the other groups were used the combination method of injecting LPS and smudging to establish COPD models. The last three groups were fed with normal saline, Qingjin Huatan Decoction, clarithromycin respectively for 30 days. On the 31st day of the experiment, the rats were put to death to take lung tissue. 6 rats from each group were chosen randomly, and the protein expressions of NE, EGFR and MUC5AC mRNA in lung tissue were detected by real-time fluorescent quantitative PCR. Results The expressions of NE, MUC5AC mRNA in lung tissue of COPD rats were higher than the healthy group. Compared with model group, the expressions of NE mRNA and MUC5AC mRNA in Qingjin Huatan Decoction group and clarithromycin group were markedly lower, while the expressions of NE, MUC5AC mRNA in Qingjin Huatan Decoction group were significantly lower than those in clarithromycin group. The expression of EGFR mRNA in Qingjin Huatan Decoction group was lower than that in model group. Conclusion Qingjin Huatan Decoction can inhibit mucus hypersecretion in airway through NE/EGFR/MUC5AC signal transduction pathway.