Morphological Effects of Cigarette Smoking on Respiratory System and Related Functions Based on AQPs Expression

SUMMARY: A great deal of attention of air pollution on respiratory health is increasing, particularly in relation to haze days. It is that exposure to cigarette smoke augments the toxicity of common air contaminants, thereby increasing the complexity of respiratory diseases. Although there are various mechanisms involved to respiratory diseases caused or worsen by cigarette smoking, in which the role of AQPs in the lung with regard to fluid homeostasis still remains elusive. In this paper, we copied the rat models based on smoke generator, and investigated the morphological changes of mucosa and related functions depending on the balance of lining liquid of alveoli via AQPs expression. Compared with normal group, weak labelling of AQP1 and AQP5 protein abundance were clearly detected in the corresponding part of smoke exposure groups compared with normal group. Hence, it is suggested that the contribution of AQPs in the lung is diminished, thereby causing perturbed balancing between resorptive and secretory fluid homeostasis under cigarette smoking.

Cada vez se presta más atención a la contaminación del aire en la salud respiratoria, particularmente, en relación con los días de neblina. En consecuencia la exposición al humo del cigarrillo aumenta la toxicidad de los contaminantes comunes del aire, lo que además aumenta la complejidad de las enfermedades respiratorias. Aunque existen varios mecanismos involucrados en las enfermedades respiratorias causadas o empeoradas por el tabaquismo, en las que el papel de las AQP en el pulmón respecto a la homeostasis de líquidos sigue siendo difícil de alcanzar. En este artículo, copiamos los modelos de rata basados en el generador de humo e investigamos los cambios morfológicos de la mucosa y las funciones relacionadas según el equilibrio del líquido de revestimiento de los alvéolos a través de la expresión de AQP. En comparación con el grupo normal, se detectó claramente un etiquetado débil de la abundancia de proteínas AQP1 y AQP5 en la parte correspondiente de los grupos de exposición al humo en comparación con el grupo control. Por lo tanto, se sugiere que la contribución de las AQP en el pulmón está disminuida, provocando así un equilibrio perturbado entre la homeostasis del líquido secretor y de reabsorción bajo el hábito de fumar cigarrillos.

mRNA Expression of Aquaporins in Human Luteinized Granulosa Cells from Different Diameter Follicles / 中山大学学报(医学科学版)

ObjectiveTo investigate the mRNA expression levels of various aquaporins （AQPs） in luteinized granulosa cells from follicles of different diameters. MethodsFrom March 25， 2022 to September 23， 2022 in our reproductive medicine center， 48 women undergoing in-vitro fertilization （IVF） were enrolled and divided into the antagonist group and the agonist group according to the ovarian stimulation protocol. Follicular fluid samples were collected on the day of oocyte pick-up and granulosa cells were extracted from follicles of different diameters： small （<13 mm）， medium （13~18 mm） and large （≥18 mm）. After RNA quantification， 22 cases （66 samples） were included for analysis and mRNA expression levels of AQPs were compared among the three follicle groups. ResultsThe mRNA expression of aquaporin 2 （AQP2） in luteinized granulosa cells increased with the increase of follicle diameter （linear trend P = 0.004） and the difference was statistically significant between two groups of large and small follicles （P = 0.017）. Statistical difference was found in the antagonist group （P = 0.049 6）， but not in the agonist group （P = 0.108）. ConclusionThe mRNA level of AQP2 in luteinized granulosa cells increases with the increase of follicle diameter and its expression is related to the ovarian stimulation protocol， suggesting that AQP2 may play a role in follicle growth and follicular fluid formation， and its mRNA expression level may be regulated by follicle stimulating hormone （FSH） and luteinizing hormone （LH）.

Luteolin ameliorates loperamide-induced functional constipation in mice

Functional constipation (FC) is one of the most common gastrointestinal disorders characterized by hard stools and infrequent bowel movements, which is associated with dysfunction of the enteric nervous system and intestinal motility. Luteolin, a naturally occurring flavone, was reported to possess potential pharmacological activities on intestinal inflammation and nerve injury. This study aimed to explore the role of luteolin and its functional mechanism in loperamide-induced FC mice. Our results showed that luteolin treatment reversed the reduction in defecation frequency, fecal water content, and intestinal transit ratio, and the elevation in transit time of FC models. Consistently, luteolin increased the thickness of the muscular layer and lessened colonic histopathological injury induced by loperamide. Furthermore, we revealed that luteolin treatment increased the expression of neuronal protein HuC/D and the levels of intestinal motility-related biomarkers, including substance P (SP), vasoactive intestinal polypeptide (VIP), and acetylcholine (ACh), as well as interstitial cells of Cajal (ICC) biomarker KIT proto-oncogene, receptor tyrosine kinase (C-Kit), and anoctamin-1 (ANO1), implying that luteolin mediated enhancement of colonic function and contributed to the anti-intestinal dysmotility against loperamide-induced FC. Additionally, luteolin decreased the upregulation of aquaporin (AQP)-3, AQP-4, and AQP-8 in the colon of FC mice. In summary, our data showed that luteolin might be an attractive option for developing FC-relieving medications.

Mechanism of Combined Therapy of Lung and Intestine in Alleviating Pulmonary Edema in Treatment of Acute Lung Injury Based on VIP/cAMP/PKA/AQPs Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo explore the mechanism of the combined therapy of lung and intestine （Mahuangtang + Da Chengqitang） in alleviating pulmonary edema in rats with acute lung injury （ALI） induced by lipopolysaccharide （LPS）. MethodWistar rats were randomly divided into blank group， model group， low-， medium-， and high-dose groups with combined therapy of lung and intestine， and positive control group. LPS （10 mg·kg-1） was given （ip） to induce ALI in rats. After modeling， the blank group was given normal saline （25 mL·kg-1）， the combined therapy of lung and intestine treatment groups were given （ig） low- （5 g·kg-1）， medium- （7.5 g·kg-1）， and high-dose （10 g·kg-1） Mahuangtang and Da Chengqitang， and the positive control group was given dexamethasone （5 mg·kg-1）. Medications were administered 0， 8， and 16 h after LPS injection for 3 times. Then lung tissue and serum were collected after administration. The lung tissues were stained with haematoxylin-eosin （HE）， and the pulmonary edema score was evaluated. The dry/wet （D/W） weight ratio of lung tissues in each group was measured， and the content of serum vasoactive intestinal peptide （VIP） in rats was detected by enzyme-linked immunosorbent assay （ELISA）. Western blot was used to detect the protein levels of aquaporin-1 （AQP1）， AQP5， VIP， cyclic adenosine monophosphate （cAMP）， phosphorylated protein kinase A （p-PKA）， and PKA in lung tissues of rats in each group. The level of VIP mRNA in lung tissues of rats was detected by real-time quantitative polymerase chain reaction （Real-time PCR）. ResultCompared with the blank group， the model group exhibited obvious lung injury， increased edema score， decreased D/W ratio （P<0.01）， declined AQP1， AQP5， cAMP， and p-PKA/PKA in lung tissues （P<0.05， P<0.01）， elevated VIP content （P<0.01）， and up-regulated levels of VIP protein and mRNA in lung tissues （P<0.05， P<0.01）. Compared with the model group， combined therapy of lung and intestine treatment groups showed alleviated lung injury， increased D/W ratio （P<0.01）， elevated AQP1， AQP5， VIP， cAMP， and p-PKA/PKA in lung tissues （P<0.05， P<0.01）， and up-regulated VIP levels in lung tissues （P<0.05， P<0.01）. ConclusionThe combined therapy of lung and intestine can alleviate ALI-induced lung tissue edema， and the mechanism may be related to the activation of the VIP/cAMP/PKA signaling pathway， which further promotes the expression of AQP1 and AQP5 and enhances the water metabolism of lung tissue.

Expression of V2R and AQPs in the renal medulla of high-fat nutritional obesity of rat / Expresión de V2R y AQPs en la médula renal de ratas con obesidad nutricional alta en grasas

Recent evidence has indicated that adipose tissue produces bioactive substances that contribute to obesity-related kidney disease, altering the renal function and structure. Eight of the AQPs are expressed in the kidney, where several of them contribute to water absorption and maintenance of body water balance. In the study, we mainly examined the localization of AQP2, AQP3 and V2R in renal medulla of Normal Diet (ND) and High-fat Diet (HFD) of rats, respectively. In renal medulla of HFD, immunolight microscopy revealed weak expression of AQP2 at the apical plasma membrane and intracellular vesicles of principal cells of the IMCD and OMCD. AQP3 and V2R expression also observed a decrease in immunolabelling in the IMCD and OMCD. It was suggested that excess lipid accumulation may lead to lipotoxicity and may be the major driver of organ dysfunction such as water reabsorption dysfunction, which may be resulted from abnormal response of rphan G-protein-coupled receptors in kidney.

La evidencia reciente ha indicado que el tejido adiposo produce sustancias bioactivas que contribuyen a la enfermedad renal relacionada con la obesidad, alterando la función y la estructura renal. Ocho de los AQP se expresan en el riñón, donde varios de ellos contribuyen a la absorción de agua y al mantenimiento del equilibrio hídrico corporal. En el estudio, examinamos principalmente la localización de AQP2, AQP3 y V2R en la médula renal de ratas con dieta normal (ND) y ratas con dieta alta en grasas (HFD). En la médula renal del grupo HFD, la microscopía electrónica de barrido reveló una expresión débil de AQP2 en la membrana plasmática apical y las vesículas intracelulares de las células principales de IMCD y OMCD. La expresión de AQP3 y V2R también observó una disminución en el inmunomarcador en IMCD y OMCD. Se sugiere que el exceso de acumulación de lípidos puede conducir a lipotoxicidad y ser el principal impulsor de la disfunción orgánica, como la disfunción de reabsorción de agua, que puede ser el resultado de la respuesta anormal de los receptores acoplados a proteína rphan G en el riñón.

Comparison of the diuretic effects of Descurainiae Semen, Coicis Semen and Plantaginis Semen / 中成药

AIM To compare the diuretic effects of Descurainiae Semen (DS),Coicis Semen (CS) and Plantaginis Semen (PS),and to observe their mechanical similarities and differences.METHODS Metabolic cage method was applied to investigating the diuretic effects of DS (2.34 g/kg),CS (7.00 g/kg) and PS (3.50 g/kg),whose diuretic mechanisms were studied by cryoscopic method,enzyme method,ion selective electrode method,ELISA and Western blot.RESULTS DS,CS and PS obviously increased saline-loaded rats' urine volume (P ＜ 0.05) and reduced their body weight (P ＜ 0.05) after administration for 7 h,which exhibited no significant effects on urine creatinine (Ucr),serum creatinine (Scr) and blood urea nitrogen (BUN)(P ＞ 0.05).DS showed its diuretic effect mainly by lowering the levels of serum Na +,atrial natriuretic peptide (ANP),brain natriuretic peptide (BNP),pulmonary AQP3,renal AQP1 and AQP2;CS showed its diuretic effect mainly by reducing the levels of serum Na +,Cl-,ANP,pulmonary AQP3,gastric AQP3,renal AQP1 and AQP2;PS showed its diuretic effect mainly by decreasing the levels of serum Na + and Cl-,pulmonary AQP3,gastric AQP3,renal AQP1 and AQP2.CONCLUSION Three medicinal materials have significant diuretic effects without obvious renal harm.DS categorized as a medicinal plant of lung channel and tropism has a great effect on netriuretic peptide system,CS categorized as a medicinal plant of spleen channel and tropism has a great effect on gastric AQP3,and PS categorized as a medicinal plant of renal channel and tropism has a great effect on renal AQPs.

Effect of lipoxin A4 on the expression of aquqporin-1,3,5 in the typeⅡpenumonocyte of rat / 中华急诊医学杂志

Objective To study the effects of Lipoxins A4(LXA4)on the expressions of aquaporin(AQP)1,3,5 in type Ⅱ pneumonocytes(ATⅡ)of rat treated with lipopolysaccharide(LPS).Method One pathogenfree male Spree Dawley(SD)rat every time.weighing 200～250 g,were used for the study.The typeⅡpenumonocytes of rats were isolated and purified,and the changes of cellular ultrastructure were observed by electron microscope in order to get the purity quotien＞90%.The type Ⅱ pneumonocytes were divided randomly into five groups,namely,vebicukun group(alcohol 0.7μL/mL),control group,LXA4 group(1×10-7mol/mL),endotoxin group(LPS 1μg/mL)and LXA4+LPS group(LXA4 1×10-7mol/mL,LPS 1μg/mL).AQP-1,3,5 mRNA of in the typeⅡpenumonocytes were assayed by using reversal transcription poly chain reaction(RT-PCR),and the expressions of AQP-1,3,5 protein were detected by using.immunohistochemistry(IHC).One each specimen,these tests were repeated for six times.ANOVA was used for statistical analysis.Results RT-PCR and IHC showed that when AT Ⅱ treated with 1 μg/mL LPS for 4 hours,the AQP-1,3,5 mRNA and the expressions of AQP-1,3,5 protein were significantly decreased in LPS group compared with control group(P＜0.01).However,the AQP-1,3,5 mRNA and the expressions of AQP-1,3,5 protein after application of LXA4 significandy increased in LPS+LXA4 group in comparison with LPS group(LPS+LXA4,AQP1:0.647±0.132,AQP3:0.900±0.856,AQP5:0.879±0.058;LPS,AQP1:0.297±0.133,AQP3:0.512±0.113,AQP5:0.647±0.110;P＜0.01).The AQP-1,3,5 mRNA and the expressions of AQP-1,3,5 protein were aignificandy increased in LXA4 group in comparison with control group(LXA4,AQP1:0.539±0.142,AQP3:0.818 4-0.176,AQP5:0.841±0.066;Blank Control,AQP1:0.518±0.139;AQP3:0.138±0.136,AQP5:0.766±0.066;P＜0.01).Conclusions AQP-1,3,5 exist in typeⅡpenumonoeyte of rata,and the LXA4 can up-regulate the mRNA and protein expressions of AQP-1,3,5 in Type Ⅱ penumonocytes of rats treated with LPS.