Modified Wuhutang Alleviates Airway Inflammation and Down-regulates Expression of Muc5AC， STAT3， NF-κB， and NLRP3 in Lung Tissues of RSV-infected Asthmatic Mice / 中国实验方剂学杂志

ObjectiveTo explore effect of modified Wuhutang on airway inflammation and expression of mucin （Muc） 5AC， signal transducer and activator of transcription 3 （STAT3）， nuclear factor kappa B （NF-κB）， and NOD-like receptor thermal protein domain associated protein 3 （NLRP3） in respiratory syncytial virus （RSV）-infected asthmatic mice. MethodSeventy male BALB/c mice of 6-8 weeks old were randomized into normal control （CON）， asthma （ovalbumin， OVA）， RSV infection-induced asthma （OVA+RSV）， high-， medium-， and low-dose （4.08， 2.04， 1.02 g·kg-1·d-1， respectively） modified Wuhutang， and dexamethasone （Dxms， 0.1 g·kg-1d-1） groups （n=10）. The model of asthma was established by sensitization and atomization inhalation with OVA. The RSV infection-induced asthma model was established by three consecutive RSV nasal infusions （1.0 × 106 PFU·mL-1， 50 μL）. Wuhutang was administrated by gavage， and Dxms by intraperitoneal injection. The CON group was given the same amount of normal saline by gavage. The mice were anesthetized with 2.5% pentobarbital sodium 24 h after the last administration， and then the lung tissue was stained by hematoxylin-eosin （HE） and Van Gieson （VG） for observation of airway inflammation. The immunohistochemical assay was employed to detect the expression of Muc5AC. Western blot was employed to determine the protein levels of phosphorylated （p）-STAT3， STAT3， p-NF-κB， NF-κB， and NLRP3. ResultCompared with the CON group， the OVA group presented airway inflammatory cell infiltration， tissue hyperemia and edema， and collagen fiber deposition. The OVA+RSV group showed severer airway inflammatory cell infiltration and tissue hyperemia and edema than the OVA group. Compared with the OVA+RSV group， modified Wuhutang alleviated the airway inflammatory cell infiltration， tissue hyperemia and edema， and collagen fiber deposition， and the high-dose group had the best performance. Compared with the CON group， the OVA group and the OVA+RSV group showed increased expression level of Muc5AC （P<0.01）. Compared with the OVA+RSV group， modified Wuhutang reduced the expression level of Muc5AC， and the reduction was significant in the high-dose group （P<0.05）. Compared with the high-dose modified Wuhutang group， Dxms lowered the expression level of Muc5AC （P<0.05）. Compared with the CON group， the OVA and OVA+RSV groups showed up-regulated protein levels of p-STAT3， p-NF-κB， and NLRP3 （P<0.05， P<0.01）. Compared with the OVA+RSV group， modified Wuhutang down-regulated the protein levels of p-STAT3， p-NF-κB， and NLRP3 （P<0.01）. Compared with the high-dose modified Wuhutang group， the Dxms group showed up-regulated levels of p-STAT3， p-NF-κB proteins （P<0.01）. ConclusionModified Wuhutang can reduce airway inflammation and down-regulate the expression of Muc5AC， p-STAT3， p-NF-κB， and NLRP3 in RSV-infected asthmatic mice， which suggests that Wuhutang reduces airway inflammation in RSV-infected asthma by regulating the STAT3/NF-κB signaling pathway.

Clinical Randomized Controlled Observation of Modified Wuhutang in Treatment of Acute Asthma in Children / 中国实验方剂学杂志

ObjectiveTo explore the clinical effect of modified Wuhutang in the treatment of children with acute asthma. MethodA total of 130 children with acute asthma were randomly divided into an observation group and a control group， with 65 cases in each group. The observation group was treated with modified Wuhutang and the control group was treated with procaterol hydrochloride for one week. The scores of primary symptoms （wheezing， cough， shortness of breath， and chest tightness） and secondary symptoms （mental status， runny nose， dry mouth， tongue texture， tongue coating， stool， etc.）， lung functions， immunoglobulin E （IgE） expression， eosinophil （EOS） count， and serum inflammatory factors， including interleukin （IL）-5， IL-6， IL-8， and IL-1β in two groups before and after treatment were compared. ResultThe data of 126 children were statistically analyzed. As revealed by the results， compared with the conditions before treatment， the scores of primary symptoms and secondary symptoms， serum levels of IL， IgE expression， and EOS count were both reduced in two groups （P<0.05）， lung functions were increased in the two groups（P<0.05）. Compared with the control group after treatment， the observation group showed decreased scores of cough and secondary symptoms （P<0.05）， and insignificant decrease in IL-1β （P<0.05）. The improvement in lung functions， IgE expression， and EOS count in observation group was equivalent to that in control group. ConclusionModified Wuhutang for treatment of acute asthma in children （phlegm-heat obstructing lung syndrome） can significantly relieve the clinical symptoms， improve lung functions， and reduce IgE， IL-5， IL-6， IL-8， IL-1β expression levels and EOS count， and its overall clinical efficacy is superior or equivalent to that of tprocaterol hydrochloride.

Wuhutang Alleviates Respiratory Syncytial Virus-induced Asthma in Mice by Inhibiting STAT3 Protein and Regulating Dendritic Cell Autophagy / 中国实验方剂学杂志

Objective:To investigate the therapeutic mechanism of Wuhutang on respiratory syncytial virus （RSV）-induced asthma in mice and its influence on the expression of signal transducer and activator of transcription 3 （STAT3） in lung tissue. Method:One hundred female BALB/c mice of SPF grade were randomly divided into a normal group and an experimental group. After successful modeling via aerosol inhalation of RSV and ovalbumin （OAV）， the mice in the experimental group were further randomized into the following seven groups： model， positive control （dexamethasone， 1.82 mg·kg^-1）， STAT3 inhibitor （STATTIC， 3.75 mg·kg^-1）， STAT3 inducer （colivelin， 1.0 mg·kg^-1）， and low-， medium-， and high-dose （1.6， 3.2， and 6.4 g·kg^-1， respectively） Wuhutang groups. The corresponding drugs were administered for two weeks， followed by the detection of airway reactivity using a small animal ventilator， the pathological changes in lung tissue， mucus secretion by goblet cells and collagen deposition in airway were observed by hematoxylin-eosin （HE）， periodic acid-Schiff （PAS） and Masson staining， the serum levels of interleukin-6 （IL-6）， IL-10， and IL-17 were detected by enzyme-linked immunosorbent assay （ELISA）. The mRNA expression levels of TGF-<italic>β</italic>₁ and <italic>α</italic>-SMA in lung tissue were detected by fluorescence-based real-time polymerase chain reaction （Real-time PCR）， autophagosomes present in lung tissue were examined by transmission electron microscopy， the protein expression levels of ATG5 and SQSTM1 in dendritic cells （DCs） and STAT3 and p-STAT3 in lung tissue were detected by Western blot. Result:The airway reactivity of the model group was enhanced in contrast to that in the model group （<italic>P<</italic>0.01）， manifested as inflammatory cell infiltration around the lung tissue， excessive metaplasia of goblet cells， and extensive deposition of airway collagen， the expression levels of serum IL-6 and IL-17 were increased （<italic>P<</italic>0.01）， while that of IL-10 declined （<italic>P<</italic>0.01）， the mRNA expression levels of TGF-<italic>β</italic>₁ and <italic>α</italic>-SMA were elevated （<italic>P</italic><0.01）， the number of autophagosomes in the lung tissue increased. The protein expression levels of ATG5， STAT3， and p-STAT were up-regulated， while that of SQSTM1 was down-regulated （<italic>P<</italic>0.01）. Compared with the model group， Wuhutang and STATTIC significantly reduced the airway hyperresponsiveness of asthmatic mice （<italic>P<</italic>0.05， <italic>P<</italic>0.01）， alleviated RSV-induced pathological changes in lung tissue， reduced the contents of serum IL-6 and IL-17 （<italic>P<</italic>0.01）， increased serum IL-10 and ATG5 in DCs （<italic>P<</italic>0.01）， down-regulated the mRNA expression levels of TGF-<italic>β</italic>₁ and <italic>α</italic>-SMA as well as the protein expression levels of SQSTM1， STAT3 and p-STAT3 （<italic>P<</italic>0.05，<italic>P<</italic>0.01）， and elevated the number of autophagosomes. Conclusion:Wuhutang relieves airway inflammation， improves airway remodeling and reduces airway hyperresponsiveness in RSV-induced asthmatic mice by inhibiting STAT3 protein and up-regulating DC autophagy in lung tissue.

Clinical Effect of Addition and Subtraction Therapy of Wuhutang Combined with Qingjin Jianghuotang to Community Acquired Pneumonia in Children with Syndrome of Phlegm Heat Closing Lung / 中国实验方剂学杂志

Objective:To discuss clinical effect of addition and subtraction therapy of Wuhutang combined with Qingjin Jianghuotang to community acquired pneumonia (CAP) in children with syndrome of phlegm heat closing lung, and to study the influence to inflammatory factors. Method:One hundred and forty patients were randomly divided into control group (69 cases) and observation group (71 cases) by random number table. Patients in two group of chidren got comprehensive symptomatic treatment measures of anti-infection, antipyretic, expectorant, antiasthmatic and respiratory support of inflammatory factors. The control group was treated with Lingyang Qingfei granules.1 g/time,3 time/day. Patients in observation group added addition and subtraction therapy of Wuhutang combined with Qingjin Jianghuotang, 1 dose/day. The courses of treatment in two groups were 7 days. And temperature, time of antipyretic, time of complete antipyretic and rate of complete antipyretic at the 7th day after treatment were recorded. And release time and disappearance time of cough, expectoration, disappearance time of pulmonary rales and treatment failure were also recorded. And before and after treatment, scores of syndrome of phlegm heat closing lung were graded, and levels of serum high sensitive C-reactive protein (hs-CRP), procalcitonin (PCT), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were detected. Result:Analyzed by rank sum test, effect in observation group was better than that in control group (Z=2.106, P<0.05), and curative effect of traditional Chinese medicine (TCM) syndrome was also better than that in control group (Z=2.119, P<0.05). Time of antipyretic, time of complete antipyretic, release time and disappearance time of cough and expectoration and disappearance time of pulmonary rales were all shorter than those in control group (P<0.01). Rate of complete antipyretic at the 7th day after treatment in observation group was 96.92%（63/65） higher than 82.81%（53/64） in control group (χ2=7.085, P<0.05). Failure rate of treatment was 9.23%（6/65） lower than 23.44%（15/64） in control group (χ2=4.775, P<0.05). And major symptom, physical sign score, minor symptom score, the total score of syndrome of phlegm heat closing lung and levels of hs-CRP, PCT, TNF-α and IL-6 were all lower than those in control group (P<0.01). Conclusion:On the basis of comprehensive anti-infection treatment, addition and subtraction therapy of Wuhutang combined with Qingjin Jianghuotang can control the clinical symptoms, and the advantages of rapid onset, rapid symptom regression, short course of disease can be found, and it can also reduce the inflammatory reaction, control the progress of the disease. The complete antipyretic rate, disease efficacy and TCM syndrome efficacy are better.