Relationship between plasma SP-A expression level and disease stage in silicosis patients / 上海预防医学

ObjectiveTo investigate the relationship between plasma surfactant protein⁃A （SP⁃A） expression level and silicosis progression， and to provide early evidence for exploring whether SP⁃A can be used as a biomarker for clinical monitoring of silicosis disease progression. MethodsWe recruited 187 silicosis patients in Guangdong Province hospital for occupational disease prevention and treatment between November， 2019 and November，2020. Their peripheral venous blood samples were collected for the plasma isolation. The level of pulmonary SP⁃A was detected by enzyme-linked immunosorbent assay. ResultsThere was a statistically significant difference in the level of SP⁃A among the silicosis groups （P<0.05）， and the plasma SP-A level of the silicosis patients in stage Ⅲ was higher than that in stage Ⅰ and stage Ⅱ （P<0.05）. Smoking had effect on plasma SP⁃A levels， Age， working years and drinking had no effect on plasma SP⁃A levels. ConclusionThe expression level of SP⁃A in the plasma of silicosis patients is increased， which has a certain correlation with the disease stage， and plays a certain early warning role in the occurrence and development of silicosis， and may be a potential biomarker for the diagnosis and prognosis of silicosis.

The role of surfactant associated protein-A in silicosis / 中国职业医学

Objective: To investigate the role of surfactant associated protein-A (SP-A) in the development and progression of silicosis, and its mechanism. Methods: Homozygous and heterozygous mice of SP-A knockout of specific pathogen free (SPF) grade were selected for mating, and mice with SP-A-/- genotype were selected for subsequent experiments. SP-A wild-type (SP-A+/+) and SP-A-/- mice were divided into SP-A+/+ control group, SP-A-/- control group, SP-A+/+ silicosis group and SP-A-/- silicosis group with six mice in each group by random number table method. Mice in both silicosis groups were given 20.0 μL 250 g/L silica suspension by tracheal exposure, and mice in both control groups were injected with 0.9% sodium chloride solution at the same volume. On the 28th day after modeling, mice were sacrificed. Lung tissues were used for lung histopathology examination. The apoptosis of alveolar type Ⅱ epithelial cells of mice was detected by TUNEL method. The mRNA expression of B-lymphoblastoma 2 (Bcl-2), Bcl-2 associated X protein (Bax), cysteinyl aspartate specific proteinase (Caspase)-3 and Caspase-9 in lung tissues of mice was detected by quantitative real-time polymerase chain reaction. Results: The histopathological result of mice showed that thickened alveolar septum, scattered silicon nodule and collagen fiber formation were observed in the mice lungs of SP-A+/+ silicosis group, and a large number of inflammatory cells were observed in silicosis nodule, after exposure to silica dust. SP-A-/- silicosis group resulted in a more severe pulmonary inflammation and interstitial fibrosis compared to SP-A+/+ silicosis group. The apoptosis of alveolar type Ⅱ epithelial cells and the mRNA relative expression levels of Bax, Caspase-3 and Caspase-9 in lung tissues of mice in each silicosis groups were increased compared with their control groups (all P<0.05). The above four indexes of mice in SP-A-/- silicosis group were higher than those in SP-A+/+ silicosis group (all P<0.05). There was no significant difference in the expression of Bcl-2 mRNA in lung tissues of these four groups (P>0.05). Conclusion: Knockout of SP-A can aggravate inflammation and pulmonary fibrosis in silicosis model mice, and promote apoptosis of alveolar type Ⅱ epithelial cells. The mechanism may be related to the Bcl-2/Bax/Caspase-3 signaling pathway which affects the apoptosis of mitochondrial pathway.

Clinical significance of Clara cell secreted protein 16 and pulmonary surfactant protein-A in early diagnosis of neonatal acute respiratory distress syndrome / 中国小儿急救医学

Objective:To investigate the clinical significance of changes of serum Clara cell secretory protein(CC16) and pulmonary surfactant protein A(SP-A) in neonates with acute respiratory distress syndrome(ARDS).Methods:The data of 30 neonates with ARDS who needed mechanical ventilation in neonatal intensive care unit of Xi′an Children′s Hospital from January 2016 to November 2018 were collected as observation group, including 12 cases in mild group, 10 cases in moderate group and 8 cases in severe group.The data of healthy newborns during the same period were taken as control group.The serum levels of CC16 and SP-A were detected by ELISA.The serum levels of CC16 and SP-A among different groups were compared.Results:The levels of serum CC16 and SP-A in ARDS group were (59.35±3.67)mg/L and(75.38±6.27)mg/L respectively, (11.26±1.32)mg/L and(18.15±2.69)mg/L in healthy group.The difference was significant( P<0.05). And the differences of serum CC16 and SP-A levels among different degree ARDS groups were significant( P<0.05). The levels of serum CC16 in mild, moderate and severe subgroup were(38.27±16.01)mg/L, (51.25±15.63)mg/L, (84.76±13.12)mg/L and SP-A were(47.02±7.18)mg/L, (73.12±7.98)mg/L, (96.45±12.50)mg/L, which increased with disease severity. Conclusion:Serum CC16 and SP-A are increased and correlated with the severity of neonatal ARDS, which may be used as the index for evaluating the severity of neonatal ARDS in the future.

Role of surfactant proteins in viral keratitis / 国际眼科杂志(Guoji Yanke Zazhi)

@#Innate immunity plays an important role in viral keratitis. Recently, it has been found that surfactant proteins(SP)A and D in the innate immune system are essential in viral keratitis. SP can inhibit virus adhesion to host cells and further promote phagocytosis of virus through high affinity for virus ligands. In order to ensure the normal function of tissues in the early stage of virus infection, SP regulates immune cells to maintain a non-inflammatory state. However, when pathogen invasion increases, SP promoted inflammation and increased the immune cells to kill the pathogens. SP-A and SP-D could be expressed in cornea, conjunctiva. To play the role of anti-adenovirus, herpes simplex virus, cytomegalovirus and other major eye pathogenic viruses, SP-A and SP-D combine with the virus to prevent entry into cells, promote phagocytosis, and directly kill the virus. SP-A and SP-D may be used as clinical diagnostic tools for viral infection. In the future, recombinant SP is expected to be used as an important means for the treatment of viral keratitis. Here, we review the innate immune function of SP-A and SP-D in ocular viral infection.

Correlation between polymorphisms of surfactant protein A1 rs1059047 and rs1136450 and respiratory distress syndrome in Mongolian premature infants / 中华急诊医学杂志

Objective To study the correlation between polymorphisms of surfactant protein A1 rs1059047 and rs1136450 and respiratory distress syndrome (RDS) in Mongolian premature infants in Inner Mongolia.Methods Totally 50 Mongolian RDS premature infants in our ward were recruited as the case group (33 males and 17 females),and another 50 Mongolian non-RDS premature infants with same ethnicity,same sex and gestational age were served as the control group (29 males and 21 females).Single nucleotide polymorphisms of SP-A1 rs1059047 and,rs1136450 and allele haploids (6A,6A2,and 6A3) were detected by polymerase chain reaction-single strand conformation polymorphism gene detection technology and were compared between the case and control groups.Results Threegenotypes,CC,TT,CT were detected in the case and control groups at rs1059047,all ofwhich were mainly TT genotype.There was no significant difference in genotype frequency between the two groups (x2=1.429,P ＞ 0.05).Two genotypes,CG and GG,were detected in the case and control groups at rsl 136450,and CG was the dominant genotype.There was no significant difference in genotype frequency between the two groups (x2=1.624,P＞0.05).The distribution frequency of SP-A1 allele haploids (6A,6A2,6A3) in the case group was 36％,68％ and 42％,respectively,and 62％,46％ and 50％ in the control group,respectively.There was no significant differencein the frequency of allele haploid 6A3 between the two groups (x2=0.502,P＞0.05);but there was a significantly difference in the frequency of allele haploids (6A,6A2) between the two groups (x2=6.763,4.937,P＜0.05).Conclusions The alleles and allele fiequency of SP-A1 (rs1059047,rs1136450) were not associated with RDS in Mongolian premature infants.However,SP-A1 allele haploid 6A2 is the susceptible gene for RDS in Mongolian premature infants,and haploid 6A is the protective gene.

Diagnostic value of lung injury biomarkers in neonatal acute respiratory distress syndrome / 国际儿科学杂志

Objective@#To investigate the role of E-selectin, Clara cell secretory protein 16 (CC-16), and pulmonary surfactant protein A (SP-A) in the diagnosis of neonatal ARDS.@*Methods@#Full-term newborn with ARDS in 9 hospitals of Jiangsu Province from March 1st 2015 to February 29th 2016 were selected as observation group.According to the lung oxygenation of the neonates, they were divided into three groups: mild, moderate and severe.In addition, 60 normal full-term newborns were selected as control group.In the observation group, venous blood samples were taken on the 1st, 3rd and 7th day of diagnosis and the control group within 7 days after birth.The level of E-selectin, CC-16 and SP-A were detected by double antibody sandwich enzyme-linked immunosorbent assay.The changes of the level of E-selectin, CC-16 and SP-A at different time points and in neonatus with different severity of ARDS were compare with control group and correlatively analyzed.@*Results@#The observation group included 60 newborns who met the diagnostic criteria of ARDS with male 38, female 22, day age (7.3 ±3.3) hours, gestational age (39.5 ±1.7) weeks and birth weight (3280 ±577) g. The control group included 60 normal full-term newborns, with male 30, female 30, day age (6.9 ±4.2) hours, gestational age (39.4 ±1.5) weeks and birth weight (3329 ±593) g. There was no significant difference between two groups.The levels of E-selectin[1 d, 3 d, 7 d: (36.36 ±8.32)μg/L, (45.51 ±9.26)μg/L, (57.15 ±6.84)μg/L], CC-16[1 d, 3 d, 7 d: (25.24 ±8.63)mg/L, (48.33±10.83)mg/L, (18.84±10.11)mg/L]and SP-A [1 d, 3 d, 7 d: (58.38±10.31)mg/L, (53.29±11.31)mg/L, (25.99±6.66)mg/L]in the blood of the observation group increased on the first day and reached the peak on the third day, which were significantly higher than those in the control group [E-selectin, CC-16, SP-A: (15.52 ±6.24)μg/L, (11.26 ±5.18)mg/L, (24.30 ±5.27)mg/L] (P<0.05). The levels of E-selectin [mild, moderate, severe are(30.07±6.10)μg/L, (33.39 ±6.64)μg/L, (41.63 ±7.36)μg/L], CC-16 [mild, moderate, severe are(12.61 ±5.80)mg/L, (25.22 ±6.77)mg/L, (30.61 ±4.69)mg/L]and SP-A [mild, moderate, severe are(49.67 ±8.26)mg/L, (7.11 ±7.94)mg/L, (63.19 ±11.45)mg/L]increased gradually in the blood of ARDS neonates with different severity (P<0.05), especially in moderate and severe degree.There was a significant negative correlation between E-selectin (r=-0.629 8), CC-16 (r=-0.679 3), SP-A (r=-0.458 8) and PaO2/FiO2 (P<0.05).@*Conclusion@#The levels of E-selectin, CC-16 and SP-A in the blood of ARDS neonates increased significantly and were closely related to the severity of the disease, which may be a biomarker of neonatal lung injury.

Serum levels of SP-A and SP-D in stage Ⅰ coal workers' pneumoconiosis and their significance / 中国职业医学

OBJECTIVE: To explore the changes of serum levels and significance of surfactant protein( SP) A and SP-D in patients with stage Ⅰ coal workers' pneumoconiosis( CWP). METHODS: A random sampling method was used to select 88 cases of stage Ⅰ CWP patients as the CWP group,50 cases of healthy underground miners with similar dust exposure history as the dust exposure group and 38 cases of ground workers without dust exposure history as the control group. The serum levels of SP-A and SP-D in 3 groups were detected by enzyme-linked immunosorbent assays. RESULTS: The levels of serum SP-A and SP-D in the CWP group and the dust exposure group were higher than that of the control group( P <0. 05). The serum level of SP-D in the CWP group was higher than that of the dust exposure group( P < 0. 01). The serum level of SP-D in the smoking CWP subgroup was lower than that of the non-smoking CWP subgroup( P < 0. 05).CONCLUSION: The abnormal serum levels of SP-A and SP-D were related to the development of stage Ⅰ CWP. Smoking might affect the serum level of SP-D in stage Ⅰ CWP patients.

The role of erlotinib in the acute lung injury and the expression of surfactant protein A / 中华急诊医学杂志

Objective To investigate the role of erlotinib in the expression of surfactant protein A (SP-A) in LPS-induced acute lung injury (ALI) of mice model.Methods C57BL/6 mice were randomly (random number)divided into control group (n=6),ER group (n=6),LPS group (n=6),and ER+LPS group (n=6).In the LPS group,2 mg/kg LPS was instilled into trachea of mice to induce lung injury.In control group,normal saline was instilled into trachea of mice instead.In the ER+LPS group and ER group,100 mg/kg of edotinib was instilled into stomach of mice,and one hour later.2 mg/kg LPS was instilled into trachea of mice in ER+PLS group to induce lung injury.Twenty-four hours later,bronchoalveolar lavage fluid (BALF) and lung tissue of mice in four groups were collected.HE staining were used for evaluating pathological changes of lung injury.Lung wet/dry weight ratio,protein concentrations and total cell numbers in the BALF were measured to determine the degree of pulmonary edema.Immunohistochemical staining and Western Blot were used for testing the protein expression of SP-A,Data of multiple groups were analyzed by one way variance (ANOVA) and inter-group comparisons were made by the least significant difference (LSD) tests.Results There was no significant difference in lung injury score (LIS) between control group (0.056±0.008) and ER (0.064±0.037) group,The LIS in LPS group (0.846-±0.047) was higher than that in control group,however the LIS in ER+LPS group (0.279±0.020) was significant lower than that in LPS group (P ＜ 0.05).Lung wet/dry weight,SP-A concentration and total cell numbers in the bronchoalveolar lavage fluid revealed that the degree of pulmonary edema in LPS group was higher than that in control group,and this pulmonary edema was reversed by erlotinib treatment.Immunohistochemical staining and Western blot showed that the expression of SP-A in LPS group was decreased compared with control group,but it was recovered after erlotinib treatment (P ＜ 0.05).Conclusions Erlotinib could protect the LPS-induced ALI,and it may be related to the regulation of SP-A.

Clinical value of surfactant protein-A in exudate pleural effusion / 中南大学学报(医学版)

Objective:To evaluate the clinical value of surfactant protein-A (SP-A) in exudate pleural effusion (EPE).Methods:This clinical study was prospective,observational and cross-sectional.Two hundred and fifteen patients with pleural effusion were divided into the transudate pleural effusions (TPE) group and the EPE group.TPE patients served as the control group.The concentrations of pleural effusions SP-A (SP-Apl) and serum SP-A (SP-Ase) were measured by ELISA,and receiver operator characteristic (ROC) curve and multivarate Cox analysis of SP-A was analysed for its clinical value.Results:SP-Apl concentrations in the EPE group were significantly higher than that in the TPE group [(189.8±43.4) ng/mLvs (22.3±5.1) ng/mL,P＜0.01];SP-Ase concentrations in the EPE group were higher than that in the TPE group [(78.9±11.3) ng/mL vs (25.8±12.4) ng/mL,P＜0.05];SP-Apl concentrations were significantly higher than the concentrations of SP-Ase in the EPE group (P＜0.01).In EPE group,SP-Apl and SP-Ase concentration in the patients with primary lung adenocarcinomas were the highest.The cut off value of SP-Apl concentrations was more than 484.5 ng/mL,yielding a 85.4％ sensitivity and 95.2％ specificity for diagnosing primary lung adenocarcinomas,with an area under the curve (AUC) of 0.943 (95％ CI 0.852 to 0.934,P＜0.01);when SP-Ase concentration was more than 84.2 ng/mL,it yielded a 76.4％ sensitivity and 94.3％ specificity for diagnosing primary lung adenocarcinomas,with an AUC of 0.910 (95％ CI 0.921 to 0.953,P＜0.01).Conclusion:While SP-Apl concentration is more than 484.5 ng/mL and/or SP-Ase concentration is more than 84.2 ng/mL,it may be helpful for the diagnosis of primary lung adenocarcinomas with the usage ofpleural effusion.

Expression of miRNA-1247-3P in acute respiratory distress syndrome cell model and its function / 实用医学杂志

Objective To detect the expression pattern of microRNA in A549 cells treated by lipopolysaccharide, study the expression of miRNA-1247-3P in A549 cells under LPS treatment and explore the possible mechanism of miRNA-1247-3P in A549 cells under LPS treatment. Methods A549 cells were divided into experimental and control groups. Immunocytochemical method and RT-PCR were used to detect the changes of SP-A and SP-C. The expression of miRNAs were detected using miRNAs array in different groups. The key miR-1247-3P was collected to detect the changes of miR-1247-3P in all groups using quantitative real-time polymerase chain reaction. Results Compared with control group, the expressions of SP-A and SP-C were significantly decreased in the experimental groups (P < 0.05). MiRNA array showed that 31 miRNAs were up-regulated and 3 miRNAs were down-regulated. Compared with control group, the expression of miR-1247-3P was significantly increased in the experimental groups (P < 0.05). Conclusion The increased expression of miR-1247-3P may play an important role in the pathogenesis of ARDS.

Effects of hyperoxia solution on pulmonary surfactant and histological changes in rabbits with oleic acid-induced acute lung injury / 西安交通大学学报(医学版)

Objective To observe the effects of hyperoxia solution on pulmonary surfactant and histology of rabbits with oleic acid-induced acute lung injury.Methods Thirty healthy rabbits weighing 2.0 - 2.5 kg were randomly divided into three groups with 10 rabbits each:control group,hyperoxia solution treatment group,and saline treatment group.Blood samples were taken for blood-gas analysis before and at 30,60 and 120 min after oleic acid or normal saline administration.Two hours later,the animals were killed and pathologic changes of lung tissue were observed microscopically.The expression of SP-A was investigated using the immunohistochemical method and image analysis system.Results PaO2 was significantly higher in hyperoxia solution treatment group than in saline treatment group two hours after treatment (P < 0.001 ),but PaCO2 was significantly lower than that in saline treatment group (P <0.001).Optical microscopy showed that lung tissue damage was milder in hyperoxia solution treatment group than in saline treatment group.The content of surfactant A in hyperoxia solution treatment group was significantly greater than that in saline treatment group (P < 0.001 ).Conclusion Hyperoxia solution can lessen lung tissue injury and damage to pulmonary surfactant.

Distribution of surfactant protein A2 haplotype and its association with preterm respiratory distress syndrome susceptibility / 中华实用儿科临床杂志

Objective To study the distribution of surfactant protein A2 (SP-A2) haplotype and its association with preterm respiratory distress syndrome (RDS) susceptibility in a local Chinese cohort.Methods Using population base and case-control study cohorts,genotyping for four single nucleotide polymorphism (SNP) was performed,rs1059046,rs17886395,rs1965707,rs1965708,in 80 term infants,50 preterm infants with RDS (RDS group) and 50 preterm infants without RDS(control group) by using TaqMan (R) real-time polymerase chain reaction.Infants in RDS group and control group were matched according to gender and gestational age.Frequency of each haplotype was compared between preterm infants with RDS and without RDS,term infants and preterm infants without RDS.Results Most common haplotypes in term infants were 1A0,1A5,1A1.In each preterm infants groups with RDS and without RDS,1A0,1 A5,1 A1 were also the most common haplotypes.Among these three common haplotypes,frequency of 1A0 was lower in preterm infants,including RDS group and control group,than that in term infants.No significant difference was found between preterm groups with RDS and without RDS (P ＞ 0.05),neither in preterm infants with gestational age ≥32 or ＜ 32 weeks.Haplotype 1A0 frequency(0.542) in term infants was significantly higher than that in preterm infants ＜ 32 weeks without RDS (0.329) (x2 =6.06,P =0.01).Conclusions SP-A2 haplotype distribution in a local Chinese population shows ethnic characteristics to some extent.Only SP-A2 does not contribute to the susceptibility for preterm RDS.

The early diagnosis value of the serum surfactant protein a in critically ill acute lung injury of HFMD / 重庆医学

Objective To explore the early diagnosis clinical value of the serum surfactant protein-A (SP-A) against acute lung injury on HFMD (hand ,foot and mouth disease) in critically ill .Methods 60 cases of HFMD were selected in Xingtai People′s Hospital from August 2010 to December 2011 ,and they were divided into three groups .20 were ordinary cases ,28 were severe cases and 12 were critical cases(4 cases dead) .According to PaO2/FiO2 of ALI ,3 of critical cases (PaO2/FiO2 >300 mm Hg) were put into the non lung injury group and 9 (PaO2/FiO2 ≤300 mm Hg) were put into the lung injury group .Besides ,15 cases of healthy children were selected as the control group .The changes of the serum SP-A levels in these children were detected through ELISA methods after 24 h and 72 h .Results Contrasting the serum SP-A levels in the ordinary and severe groups separately with the ones in control group ,there was no statistical significance(P>0 .05) and so was contrasting the serum SP-A levels in the ordinary group with the ones in the severe group ,and the serum SP-A levels in the critical group after 24 h was significantly higher than the ordina-ry and severe groups (P0 .05) ,con-trasting with ones in the control group ;but the serum SP-A levels in the lung injury group after 24 h were significantly higher than ones in the control group and in the non lung injury group (P<0 .05) .Conclusion Detection of the serum SP A has clinical value of the early diagnosis of acute lung injury on HFMD in critically ill ,which is beneficial to guide the clinical treatment .Meanwhile , it can reduce the mortality rate and the sequela ,and help to diagnose the condition of acute lung injury and treat it .

Effect of hyperoxia on A549 cells suppressed with surfactant protein-A suppressed by small RNA interference-mediated gene silencing / 中华实用儿科临床杂志

Objective To explore the effect of hyperoxia on A549 cells suppressed with surfactant protein A (SP-A) suppressed by small interference RNA(SiRNA)-mediated gene silencing,and discuss the function of SP-A in hyperoxic lung injury.Methods A549 cells were gained by serial sub cultivation in vitro and randomly divided into 2 groups,silenced of SP-A group and the control group.A549 cells were transfected with synthetic SP-A sequence-specific SiRNA by Lipofectamine 2000,continuously exposed to hyperoxia(950 mL/L 02,50 mL/L CO2).After exposure to hyperoxia for 48 hours,72 hours and 96 hours,total protein and culture supernatant were gained.SP-A protein was detected by Western-blot,the capacity of proliferation was detected by methyl thiazolyl tetrazolium,and thiobarbituric acid colorimetric method was used to detect the malondialdehyde (MDA) in culture supernatant.Results Sequence-specific SiRNA targeted SP-A2 and significantly down-regulated its expression in A549 cells.Compared with the control group in hyperoxia,the expression of SP-A significantly decreased after 48 hours,72 hours in the silenced group (all P ＜ 0.05),and the capacity of proliferation in A549 cells silenced by SP-A were obviously decreased after 48 hours,72 hours and 96 hours(all P ＜0.05).But there was no significant difference in the MDA in culture supernatant between 2 groups(all P ＞ 0.05).Conclusions The capacity of resisting hyperoxia decreased in A549 cells silenced by SP-A,which indicates that SP-A can protect hyperoxic lung injury.

Innate immunity of surfactant protein A and D in urinary tract infection with uropathogenic Escherichia coli / 中华肾脏病杂志

Objective To investigate the role of surfactant protein (SP)-A and SP-D in urinary tract infection mouse model,and evaluate the effects of SP-A and SP-D absence on urinary tract infection.Methods SP-A and SP-D double knockout (SP-A/D KO) mice were made.SP-A/D KO and wild-type (WT) C57BL/6 female mice were used for this study.The expression of SP-A and SP-D in kidney was detected by immunohistochemistry (IHC).The levels of p-p38 and p38 protein in kidneys were measured by Western blotting.Uropathogenic Escherichia coli or buffer was delivered into the bladder of female mice.At 24 and 48 h after inoculation,CFU of Escherichia coli in the kidney and urine of the treated and control mice were measured.Histological,cellular and molecular analysis were performed by several methods of H/E staining,IHC and Western blotting.The effects of SP-A and SP-D on bacterial growth were studied in vitro.Results SP-A and SP-D in kidney were located in the proximal tubules and collecting tubules.Compared with WT mice,infected SP-A/D KO mice with UPEC had higher CFU in kidneys and urine at 24 h and 48 h,increased inflammatory cells infiltration in kidneys (P＜0.05).Compared with WT mice,SP-A/D KO mice had higher p38 MAPK phosphorylation levels in kidneys (P ＜ 0.05).Growth of Escherichia coli was greatly inhibited by both SP-A and SP-D (P＜0.05).Conclusions Both SP-A and SP-D are expressed in kidney.SP-A and SP-D can attenuate UTI induced by UPEC which may be through inhibiting bacterial growth and modulating renal inflammation.

Decreased Expression of Surfactant Protein Genes Is Associated with an Increased Expression of Forkhead Box M1 Gene in the Fetal Lung Tissues of Premature Rabbits

PURPOSE: Recently, Forkhead box M1 (FoxM1) was reported to be correlated with lung maturation and expression of surfactant proteins (SPs) in mice models. However, no study has been conducted in rabbit lungs despite their high homology with human lungs. Thus, we attempted to investigate serial changes in the expressions of FoxM1 and SP-A/B throughout lung maturation in rabbit fetuses. MATERIALS AND METHODS: Pregnant New Zealand White rabbits were grouped according to gestational age from 5 days before to 2 days after the day of expected full term delivery (F5, F4, F3, F2, F1, F0, P1, and P2). A total of 64 fetuses were enrolled after Cesarean sections. The expressions of mRNA and proteins of FoxM1 and SP-A/B in fetal lung tissue were tested by quantitative reverse-transcriptase real-time PCR and Western blot. Furthermore, their correlations were analyzed. RESULTS: The mRNA expression of SP-A/B showed an increasing tendency positively correlated with gestational age, while the expression of FoxM1 mRNA and protein decreased from F5 to F0. A significant negative correlation was found between the expression levels of FoxM1 and SP-A/B (SP-A: R=-0.517, p=0.001; SP-B: R=-0.615, p<0.001). CONCLUSION: Preterm rabbits demonstrated high expression of FoxM1 mRNA and protein in the lungs compared to full term rabbits. Also, the expression of SP-A/B was inversely related with serial changes in FoxM1 expression. This is the first report to suggest an association between FoxM1 and expression of SP-A/B and lung maturation in preterm rabbits.

Surfactant Protein A Polymorphism Is Associated with Susceptibility to Chronic Obstructive Pulmonary Disease in Chinese Uighur Population / 华中科技大学学报（医学）（英德文版）

This study investigated the correlation between surfactant protein-A (SP-A) polymorphism and the susceptibility of chronic obstructive pulmonary disease (COPD) in Xinjiang Uighurs.Genomic DNA was extracted from peripheral blood of 194 COPD smokers and 201 healthy smokers of Uighur who were hospitalized in or paid a visit to one of the four Xinjiang-based hospitals involved in the study,from March 2009 to December 2010.Single nucleotide polymorphisms (SNPs) were studied at aa62 (CCA/CCG rs 1136451) and aa219 (CGG/rGG,rs4253527) in SP-A.Genotypes were determined by using the TaqMan polymerase chain reaction (PCR).Our results showed that genotype frequencies were different between the COPD and normal smokers in aa62 (x2=6.852,P=0.033).There were also significant differences in allele genotype frequencies between the COPD and the control and allele G might decrease the risk COPD (x2=6.545,P=0.011; OR=0.663; 95％ CI:0.484-0.909).The result suggested that polymorphism of aa62 (CCA/CCG,rs1136451) of SP-A may be associated with the susceptibility to COPD in Xinjiang Uighurs.

Protective effect of anti-inflammatory peptides on LPS-induced acute lung injury of mouse / 中国小儿急救医学

Objective To explore the anti-inflammatory effect of antiflammin-2 (AF2) and recombinant peptide sequence 2(R2) on acute lung injury of mouse. To observe the expression of clara cell 16000 protein (CC16) and surfactant protein A (SP-A) in the lung of mouse inoculated with lipopolysaccharide (LPS) and the impact of AF2,R2,and glucocorticoids(hydrocortisone,HC) may have on the expression of the CC16 and SP-A in the lung of mice with acute lung injury. Methods Balb/c mice were inoculated with LPS (5 mg/kg) by intraperitoneal injection to set up ALI mice model. Mice weighed from 15 g to 16 g were grouped into control group, model group and treated groups respectively treated with AF2, R2 or HC. Mice in the control group were injected with physiological saline solution, while mice in the other four groups were inoculated with LPS to induce acute lung injury. Then animals in the treated groups were treated with AF2, R2 or HC each on a dose of 2 mg/kg also through intraperitoneal injection,while those of the control group and the model group, were given equivalent physiological saline solution as a placebo. The respiratory rate of all of these animals were recorded 6 hours after the injection. And at the time point of 12 hour,all the mice were sacrificed for a preparation of the whole lung tissue for the sake of a pathological investigation ,or for extractions of RNA for a semiquantitative analysis of the expression of CC16 and SP-A within the lungs. Results (1) An obvious attenuation of the respiratory rates of the three treated groups were observed when comparing with that of the mice in the model group without any anti-inflammatory treatment. (2) Remarkable extenuation of the extent of intra-alveolar and intersticial hemorrhage and infiltration of inflammatory cells were observed within the treated groups comparing with that of the model group. (3) An attenuate expressions of CC16 or SP-A were observed in the model group,while obvious uptrend of CC16 expression was observed in AF2 treated groups and increase of SP-A expressions were found in R2 and HC treated groups. Conclusion The anti-inflammatory effect of the peptide, AF-2 or R2, has been conformed on ALI mice model induced by LPS.

Effect of antioxidant on pulmonary surfactant in acute lung injury rats / 中国小儿急救医学

Objective To explore the changes of neutrophil elastase (NE) and surfactant protein A (SP-A) in acute lung injury(ALI) rats,and the effect of antioxidant. Methods Sixty healthy mature Wister rats were divided into 2 groups, the control group and treatment group. The rats in two groups all received peritoneal injection of E. coli to establish the ALI animal model. 30 minutes after injection of E. coli,the rats in treatment group were injected reduced glutathione from vena caudalis. The levels of NE in blood and expressions of SP-A in lung tissue were detected at 3,6 and 12 hours after injection of E. coli. Results ALI symptom appeared 3 hours after injection of E. coli in the control group, obvious after 6 hours, the rats vomi-ted pink secretion after 12 hours. Lung edema and bleeding were found by pathologic examination. No obvious symptom was found in treatment group after 3 hours, slight tachypnea after 6 hours, slight edema in pulmonary tissue after 12 hours. After administration of reduced glutathione,levels of NE at 3,6 and 12 hours in the treatment group were lower than those in the control group,and indicated statistical significance in 6 and 12 hours(P <0. 05) ;Levels of SP-A in 3,6 and 12 hours in the treatment group were higher than those in the control group, and indicated statistical significance in 3,6 and 12 hours (P < 0. 05). Conclusion Dysfunction of pulmonary surfactant is secondary in ALI, degradation of SP-A is the one of reasons, the application of reduced glutathione as antioxidant, could effectively suppress NE to decompose basosexine elastin of cells and destroy surface active protein, has protective effect on ALI.

Lipopolysaccharide-induced expression of surfactant protein A subtypes in human renal tubular epithelial cells / 中华肾脏病杂志

Objective To determine the surfactant protein A (SP-A) subtype distribution and expression in human renal tissue and cultured human renal tubular epithelial cells(HK-2), and to explore the influence of Lipopolysaccharide (LPS) on the expression of SP-A subtypes mRNA and SP-A protein. Methods lmmunohistochemical staining was performed using SP-A polyclonal antibodies. RT-PCR was performed with mRNA from HK-2 cells and normal human kidney.Restriction fragment length polymorphism(RFLP) and sequencing were used to evaluate the subtypes of SP-A. The relative content of SP-A mRNA in human kidney and human lung was compared by real-time PCR. Western blotting analysis for SP-A was performed on protein from renal tissue and cultured HK-2 cells SP-A protein in human urine and culture supernatant of HK-2 cells was measured by enzyme-linked immunosorbent assay (ELISA) and Western blotting respectively. HK-2cells were treated with LPS at various concentrations (0,0.1,1,2,5,10 mg/L) for 8 h and at 5mg/L for various time points (0,2,4,8,16,24 h). Expression of SP-A mRNA and protein was analyzed by RT-PCR and Western blotting. Results SP-A was localized in renal tubular epithelial cells of both proximal and distal convoluted tubules. SP-A1, SP-A2 mRNA and protein could be detected in normal HK-2 cells and human kidney. The significant secretion of SP-A [urine: (106.614172.772) nmol/L, n=30; culture supernatant: (85.533±58.622) nmol/L, n=10] was shown. The levels of SP-A1, SP-A2 mRNA and Sp-A protein in HK-2 cells were significantly decreased after treatment with LPS. Conclusions Human renal tubular epithelial cells can express both SP-A1 and SP-A2 genes which may play an important role in inflammation modulation of kidney.