Clinical And Experimental Study Of Dachengqi Decoction Based On "Lung Being Relative To The Large Intestine " In Treating Lung Injury Caused By Sepsis

Objective:Based on the theory of "lung being relative to the large intestine",Dachengqi Decoction(DCQD)was used to treat patients with sepsis and lung injury.The relevant indicators and prognostic indicators of lung injury severity were monitored to observe the protective effect of DCQD on lung injury in patients with sepsis.To investigate the effect of DCQD on inflammatory cytokine production and AQP-1/AQP-5 protein expression in lipopolysaccharide(LPS)-induced ALI model and its potential mechanism.Method:1.From December 2017 to December 2018,the patients who were admitted to the Department of Acute and Critical Care Medicine of Jiangsu Provincial Hospital of Integrated Traditional Chinese and Western Medicine for the diagnosis of sepsis with lung injury reached the diagnostic criteria of ARDS,and conformity with the criteria of gasp syndrome and Yangming Fushi syndrome in TCM.All patients underwent endotracheal intubation or tracheotomy for mechanical ventilation.(The initial ventilation mode was volume controlled ventilation,tidal volume was 6-8 ml/kg,adjusted inhaled oxygen concentration(FiO2)and PEEP guaranteed more than 90%of Sp02).A total of 40 patients were enrolled in the study.The patients were randomly divided into DCQD group and control group(20 patients in the treatment group,7 females,13 males,with an average age of 65.35±10.22 years;20 patients in the control group).There were 11 females and 9 males with an average age of 62.00±12.81 years old.Both groups of patients were given no differential conventional treatment,including fluid resuscitation,vasoactive drugs,antibiotics,nutritional support,regulation of internal environment stability,and organ function support.Patients in the treatment group were given DCQD nasal feeding on a routine basis.After DCQD syrup,it was taken twice a day in the afternoon.The control group received an equal amount of warm water control.The therapeutic effect was observed after 1 day of treatment,and the total course of treatment was 5 days PO2/FiO2,P(Aa)O2 in blood gas analysis every 6 hours,recorded or calculated according to respiratory related parameters(static compliance=tidal volume/(platform pressure-positive end expiratory pressure)),data The mean daily value was used;the ultrasound extravascular lung water score(LUWS)was performed on days 1,3,and 5 after enrollment;the improvement of lung injury severity was compared between the two groups.At the 1st,3rd,and 5th day after enrollment,central venous blood TNF-a,IL-1,IL-6,and CRP were taken,and the improvement of inflammation index between the two groups was compared.The two groups of patients were assessed by the attending physician with the qualification of TCM practitioners before treatment and after 5 days treatment.According to the patients' clinical symptoms,signs,tongues and pulse conditions,the TCM syndrome scale were scored,and there was no statistical difference between the two groups.There were no significant differences in mechanical ventilation time,ICU time,hospital stay,and 28-day mortality between the groups.2.30 SD male rats were randomly divided into 3 groups(10 in each group),control group(Control group),LPS group and LPS+Dachengqi decoction group(DCQD group).First,intraperitoneal inj ection of LPS to prepare a rat model of acute lung injury with sepsis.The blood gas of the abdominal aorta was analyzed by arterial blood gas.The Pa02/Fi02 index of the rats in each group was compared.The lung tissue of the rats was obtained and compared.Lung weight/body weight ratio;real-time quantitative PCR was used to detect the expression of IL-6,IL-8 and TNF-a mRNA in lung tissue,and Western blotting was used to detect AQP-1 and AQP-5 protein in lung tissue.The expression of IL-6,IL-8,TNF-a mRNA,AQP-1 and AQP-5 in the lung tissues of each group were compared.3.The optimal drug concentration of DCQD on HULEC-5a cells was detected by CCK-8 assay,and the optimal LPS concentration of HULEC-5a cells was selected according to the degree of decrease of AQP-1 expression.HULEC-5a cells were cultured in vitro and randomly divided into 3 groups(control group,LPS group,LPS+DCQD group).The expression of AQP-1 and AQP-5 protein in HULEC-5a cell culture medium was detected by Western Blotting method.The levels of IL-6,IL-8 and TNF-a in each group were compared by sandwich ELISA.TLR4 protein expression was inhibited by siRNA,HULEC-5a cells were treated with TLR4 siRNA(50 nM)and control TLR4 for 36 hours,and each group(control group,LPS group,TLR4 siRNA+LPS group,DCQD+TLR4 siRNA+LPS group)HULEC-5a cells were detected.The levels of AQP-1,AQP-5,IL-6,IL-8,and TNF-a were studied to investigate the effect of DCQD on TLR4 signaling pathway in LPS-induced HULEC-5a cells.Western Blotting was used to detect IkB-a,p-IkB-a levels and p50/p65 nuclear translocation in each group(control group,LPS group,LPS+DCQD group),to study The effect of the NF-?B pathway of DCQD in LPS-induced HULEC-5a cells.Result:1.Clinical research part:The severity of lung injury related indicators:The ultrasound extravascular lung water score(LUWS)of both groups decreased after treatment,and the score improved after 3 days of treatment was not obvious,but showed a significant downward trend after 5 days.At the same time,the score of DCQD group was more obvious than that of the control group,and the difference was statistically significant(P=0.03).Cstat,P02/Fi02,P(Aa)02 and other indicators showed similar trends,and all improved significantly after the fifth day.The DCQD group index was more obvious than the control group,and the difference was statistically significant(P value was 0.03 7,0.004,0.028,respectively).Inflammatory index:TNF-a and IL-6 in both groups showed a significant decrease on the third day,and the decrease in the DCQD group was more significant.Compared with the control group,the difference was statistically significant(P=0.026,0.044,respectively).CRP and IL-1 did not change significantly on the third day,and there was no statistical difference between the two groups(P>0.05).After 5 days of treatment,TNF-a,IL-6,CRP,IL-1 decreased significantly,while the decrease in DCQD group was more significant.There were statistically significant differences between the two groups(P<0.001,<0.001,0.007,0.004,respectively).TCM symptom scale score:There was no significant difference in the scores of TCM syndrome scales between the two groups before treatment(P=0.947).After 5 days of treatment,the TCM syndrome scale scores decreased significantly,while the DCQD group decreased more significantly.There was a statistically significant difference between the groups(P=0.002).Prognostic indicators:In the prognostic indicators,the mechanical ventilation time,hospital stay,DCQD group was significantly shorter than the control group,the difference between the two groups was statistically significant(P=0.038,0.032,respectively).However,there was no significant difference between the two groups in the ICU time and 28-day mortality(P=0.268,1 respectively).2.Experimental research section:1)The PaO2/FiO2 index of rats with LPS induced by intraperitoneal injection of LPS decreased significantly,while the lung dry/wet weight ratio decreased,and the lung weight/body weight ratio increased;DCQD significantly decreased the Pa02/Fi02 index of LPS-induced rat,increases the lung dry/wet weight ratio and lowers the lung weight/body weight ratio.The mRNA levels of IL-6,IL-8 and TNF-a in lung tissue of LPS rats were significantly increased.DCQD could significantly inhibit the mRNA expression of IL-6,IL-8 and TNF-a in LPS-mediated lung tissues.The APQ-1 and APQ-5 proteins in LPS rats were significantly decreased.DCQD could increase the expression of APQ-1 and APQ-5 in LPS-mediated lung tissues.2)The expression of AQP-1 and AQP-5 in LPS-induced HULEC-5a cells decreased significantly,and the levels of IL-6,IL-8 and TNF-a increased significantly.DCQD can increase the expression of AQP-1 and AQP-5,decreased IL-6,IL-8 and TNF-a levels in HULEC-5a cells induced by LPS.LPS-inhibited AQP-1 and AQP-5 protein expression was significantly increased in TLR4 siRNA cells,whereas DQCD could not further increase AQP-1 and AQP-5 protein expression.LPS-induced phosphorylation and degradation of IkB-a in HULEC-5a cells was evident,while DCQD inhibited LPS-induced phosphorylation and degradation of IkB-a in HULEC-5a cells.LPS increased the nuclear translocation of p50/p65;while DCQD treatment significantly reduced the nuclear translocation of p50/p65 in LPS-induced control cells.Conclusion:DCQD can improve pulmonary vascular permeability and reduce lung inj ury in patients with sepsis,and has a clear lung protection.Its mechanism of action is related to inhibition of TLR4/NF-kB signaling pathway,increase of protein expression of AQP-1 and AQP-5,inhibition of inflammatory cytokine production,and reduction of pulmonary edema.