ABSTRACT
Objective To investigate the preventive effect of Xuebijing injection on acute lung injury induced by cardiopulmonary bypass (CPB) and the underlying mechanism. Methods ① In vivo experiment: 30 Sprague-Dawley (SD) rats were randomly divided into sham group, CPB group, Xuebijing pretreatment group (XBJ+CPB group) with 10 rats in each group. CPB model was reproduced in rats; and CPB was not performed in sham group, but only through arteriovenous puncture. In the XBJ+CPB group, 4 mL/kg Xuebijing injection was injected intraperitoneally 2 hours before CPB, sham group and CPB group were injected with equal volume of normal saline at the same time. The blood from femoral artery was analyzed 4 hours after operation, and the oxygenation index (PaO2/FiO2) was calculated. Then the rats were sacrificed to collect bronchoalveolar lavage fluid (BALF), and the lung permeability index (PPI) was calculated. The lung tissues were harvested, and the wet/dry weight ratio (W/D) of lung tissue was measured. The index of quantitative evaluation of alveolar injury (IQA) was measured. The levels of interleukins (IL-1, IL-6) and tumor necrosis factor-α(TNF-α) in lung tissue and BALF were measured by enzyme-linked immunosorbent assay (ELISA). The content of malondialdehyde (MDA) and the activities of myeloperoxidase (MPO) and superoxide dismutase (SOD) in lung tissue were detected by biochemical method. The microRNA-17-5p (miR-17-5p) expression in lung tissue was determined by quantitative reverse transcription-polymerase chain reaction (RT-qPCR).② In vitro experiments: type Ⅱ alveolar epithelial cells (AECⅡ) were cultured in vitro, and they were randomly divided into control group (the cells were treated by preoperative serum of CPB in patients with ventricular septal defect), CPB group (the cells were treated by serum after CPB in patients), and XBJ+CPB group (Xuebijing injection 10 g/L+serum after CPB in patients). After 12 hours of culture in each group, the expression of miR-17-5p was detected by RT-qPCR. AECⅡ cells were transfected with miR-17-5p mimic, inhibitor or corresponding control oligonucleotide (negative control), respectively, to observe the effect of miR-17-5p on Xuebijing regulating CPB-induced apoptosis rate and caspase-3 activity. Results ① In vivo experiment: compared with the sham group, the PPI, lung W/D ratio, IQA, and IL-1, IL-6, TNF-α in lung tissue and BALF, as well as MDA content and MPO activity in lung tissue were significantly increased, PaO2/FiO2 and SOD activity in lung tissue were significantly decreased. The parameters of the XBJ+CPB group were significantly improved, suggesting that Xuebijing pretreatment could improve CPB-induced ALI in rats. The expression of miR-17-5p in lung tissue of the CPB group was significantly down-regulated as compared with sham group (2-ΔΔCt: 0.48±0.13 vs. 1.00±0.11, P < 0.05);while the expression of miR-17-5p in the XBJ group was significantly up-regulated as compared with the CPB group (2-ΔΔCt: 1.37±0.09 vs. 0.48±0.13, P < 0.05), indicating that the improvement of Xuebijing injection on lung injury after CPB might be related to miR-17-5p. ② In vitro experiment: the changes in miR-17-5p expression in each group of AECⅡ cells confirmed in vivo results. After transfection of miR-17-5p mimic, the apoptotic rate and caspase-3 activity of each group were significantly lower than those transfected with negative control, and the decrease was more significant in the XBJ+CPB group [apoptotic rate: (7.37±0.95)% vs. (12.60±1.90)%, caspase-3 (A value): 0.82±0.09 vs. 1.37±0.08, both P < 0.05]. After transfection of miR-17-5p inhibitor, the apoptotic rate and caspase-3 activity of each group were significantly more than those transfected with negative control [in the XBJ+CPB group: apoptotic rate was (16.30±1.86)% vs. (12.60±1.90)%, caspase-3 (A value) was 1.78±0.13 vs. 1.37±0.08, both P < 0.05]. This indicated that the apoptosis of AECⅡ cells cultured in serum after CPB was significantly reduced by miR-17-5p, and further reduced by the pretreatment with Xuebijing. Conclusions Xuebiing injection can reduce the inflammatory reaction and oxidative stress of lung tissue in rats with ALI induced by CPB, and improve oxygenation. The mechanism may be related to up-regulation of miR-17-5p expression in AECⅡ cells and inhibition of apoptosis of AECⅡ cells.
ABSTRACT
Objective To investigate the effect of Xuebijing injection on inflammatory factors in patients with Stanford B aortic dissection (AD) after endovascular repair and approach its clinical significance.Methods Sixty patients with Stanford type B AD for endovascular repair admitted to the Department of Cardiothoracic Surgery of the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine (TCM) from January 2014 to June 2016 were enrolled, and they were divided into a Xuebijing group and a control group according to the random number table method, 30 cases in each group. The patients of Xuebijing group received 100 mL Xuebijing injection+ 50 mL normal saline intravenous drip during operation, while the patients of control group received an equal volume of normal saline, 2 times a day in both groups for consecutive 3 days. Peripheral venous blood was collected before and after treatment for 1, 2, and 3 days, and the levels of C-reactive protein (CRP), tumor necrosis factor-α(TNF-α), interleukin-6 (IL-6) were measured by enzyme-linked immunosorbent assay (ELISA), and the prognosis of the two groups was observed.Results With the prolongation of time, the postoperative levels of CRP, TNF-α and IL-6 in the two groups showed a tendency of first increase and then decrease compared with the preoperative levels, the above indexes of the two groups were all significantly higher, CRP (mg/L) and TNF-α (ng/L) peaked on the 2nd day postoperatively (control group: CRP was 146.34±13.92 and TNF-α was 72.22±7.63, Xuebijing group: CRP was 114.92±9.89 and TNF-α was 53.44±6.80, allP < 0.05), however, IL-6 (ng/L) peaked on the 1st day postoperatively (control group: 146.08±10.29, Xuebijing group: 117.88±8.84), then decreased, all reached the valley on the 3rd day postoperatively (control group: CRP, TNF-α, IL-6 was 112.59±8.54, 43.73±4.10, 70.03±4.66 respectively, Xuebijing group CRP, TNF-α, IL-6 was 87.75±7.67, 39.43±4.63, 56.65±3.27, respectively), and at the same time point, the level of CRP, TNF-α, IL-6 in the Xuebijing group were significant lower than those in the control group, the change were the most significant on 3 days after operation [CRP (mg/L): 87.75±7.67 vs. 112.59±8.54, TNF-α (ng/L): 39.43±4.63 vs. 43.73±4.10, IL-6 (ng/L): 56.65±3.27 vs. 70.03±4.66, P < 0.05]. There were no serious complications such as renal failure, severe hypoxemia, infection, paraplegia, stent shift and so on, no hospital death occurred and all patients were discharged in rehabilitative condition. Conclusions Endovascular repair in patients with Stanford type BAD may activate an inflammatory response inducing the release of a large amount of inflammatory factors during the early postoperative period, Xuebijing injection can inhibit the inflammatory reaction and prevent the occurrence of postoperative complications.
ABSTRACT
Objective:To clone and characterize the 16S rRNA of six species in the bacteria infecting respiratory tract to make gene chip.Methods:The primers of the target gene were designed and synthesized,and then the aimed fragment of the 16s rRNA was amplified by PCR and cloned.Finally the recombinant plasmids were characterized.Results:(1)The 16s rRNA gene of six species of bacteria was amplified.It was found that the size of amplified product by PCR was 1 300 bp in E.coli,S.aureus,S.pneumoniae,K.pneumoniae and H.influenzae and that of 1 100 bp in P.aeruginosa.(2)The JM109 transferred by the recombinant plasmid pMD18-T grew in Ampr culture was white colonies.(3)The specific bands could be found by restriction endonuclease and PCR analysis. (4)The sequence of the six bacterial 16s rRNA showed the same as those in the GenBank.Conclusion:The 16s rRNA of six species of bacteria is successfully amplified and cloned into plasmid pMD18-T. It will provide the basis for making gene chip detecting the six species of bacteria infecting respiratory tract.