Papel dos receptores farmacológicos AT1 e AT2 da Angiotensina II em doença periodontal induzida: aspectos moleculares / Role of pharmacological receptors AT1 and AT2 of Angiotensin II in caused periodontal disease: molecular aspects

O sistema renina angiotensina (SRA) é de grande importância para o equilíbrio hídrico e regulação da pressão arterial do organismo, além de estar associado ao estimulo de vias próinflamatórias. Seu principal peptídeo é a angiotensina II, que interage principalmente com os receptores do tipo 1 (AT1) e do tipo 2 (AT2). Foi encontrado interrelação entre as doenças cardiovasculares e a periodontite. Este estudo teve como objetivo avaliar os aspectos moleculares em camundongos submetidos a um modelo experimental de periodontite, observando a influência dos receptores de Ang II tipo 1 (AT1(-)) e Ang II tipo 2 (AT2(-)) na periodontite. Métodos: A periodontite experimental foi induzida colocando-se uma ligadura com fio de nylon 5.0 ao redor do segundo molar superior esquerdo de camundongos knockoutAT1(-), AT2(-) e selvagem (WT), subdivididos 2 grupos para cada linhagem: sem ligadura e ligadura, totalizando seis grupos: três controles e três experimentais. Após 15 dias da indução da doença os animais foram submetidos à eutanásia. Com o intuito de avaliar se as variações genéticas teriam influência sobre a periodontite foram realizadas as análises de citocinas, peptídeos e enzimas foram analisados a partir de tecidos gengivais por ELISA e RT-PCR. Resultados: Os animais WT e AT2(-) apresentaram resultados semelhantes em relação às citocinas IL-1ß, IL-6, TNF-α, com aumento dos níveis em relação aos saudáveis (p < 0,001). Houve diferenças significativas em IL-ß entre os grupo AT1(-)-L e WT-L (p < 0,05), e em IL-6 e TNF-α os grupos AT1(-)-L apresentaram diferenças significativas (p < 0,001) tanto quando comparado aos grupo WT-L quanto aos grupos AT2(-)-L. Os níveis de IL-10 foram maiores em WT-L (p < 0,01), enquanto os grupos AT2(-) e AT1(-) não apresentaram alterações significativas em relação a essa citocina. Houve diferenças significativas em Angiotensina II entre os grupos AT2(-)-NL e AT2(-)-L (p < 0,01); e em Angiotensina 1-7 entre os grupos AT1(-)-L e AT2(-)-L (p < 0,05). Para TLR2 houve diferenças entre os grupos WT-NL/WT-L (p < 0,05); AT1(-)-NL/AT1(-)-L (p < 0,01) e AT2(-)-NL/AT2(-) - L (p < 0,01). Para o receptor MAS houve diferenças entre os grupos WT-NL/WT-L (p < 0,001) e AT2(-)-NL/AT2(-)-L (p < 0,001), e também em relação ao grupo WT-L/AT1(-)-L (p < 0,001) e AT1(-)-L/AT2(-)-L (p < 0,001). Para a expressão dos peptídeos ECA e ECA2, houve diferença estatística apenas para ECA entre os tipos de grupos WT-NL/WT-L (p < 0,001). Conclusão: Os animais do grupo AT1(-) apresentaram menor inflamação que as demais linhagens doentes, assim como uma menor expressão do receptor Mas e Ang 1-7. Além disso os animais dos grupos WT e AT2(-) demonstraram resultados próximos em diversas análises, evidenciando que o bloqueio do receptor AT1, sobre os efeitos moleculares, é mais positiva (AU).

The renin angiotensin system (RAS) is of great importance for water balance and regulation of blood pressure in the body, in addition to being associated with the stimulation of proinflammatory pathways. Its main peptide is angiotensin II, which interacts mainly with type 1 (AT1) and type 2 (AT2) receptors. An interrelationship was found between cardiovascular diseases and periodontitis. This study aimed to evaluate the molecular aspects in mice subjected to an experimental model of periodontal disease, observing the influence of Ang II type 1 (AT1(-)) and Ang II type 2 (AT2(-)) receptors on periodontitis. Methods: Experimental periodontitis was induced by placing a ligature with 5.0 nylon thread around the upper left second molar of AT1(-), AT2(-) and wild-type (WT) knockout mice, subdivided into 2 groups for each strain: without ligation and ligation, totaling six groups: three controls and three experimental. After 15 days of disease induction, the animals were euthanized. In order to evaluate whether genetic variations would have an influence on periodontal disease, analyzes of cytokines were carried out, peptides and enzymes were analyzed from gingival tissues by ELISA and RT-PCR. Results: WT and AT2(-) animals showed similar results in relation to the cytokines IL-1ß, IL-6, TNF-α, with increased levels compared to healthy ones (p < 0.001). There were significant differences in IL-ß between the AT1(-)-L and WT-L groups (p < 0.05), and in IL-6 and TNF-α the AT1(-)-L groups showed significant differences (p < 0.001) both when compared to the WT-L and AT2(-)-L groups. IL-10 levels were higher in WT-L (p < 0.01), while the AT2(-) and AT1(-) groups did not show significant changes in relation to this cytokine. There were significant differences in Angiotensin II between the AT2(-)-NL and AT2(-)-L groups (p < 0.01); and in Angiotensin 1-7 between the AT1(-)-L and AT2(-)-L groups (p < 0.05). For TLR2 there were differences between the WT-NL/WT-L groups (p < 0.05); AT1(-)-NL/AT1(-)-L (p < 0.01) and AT2(-)-NL/AT2(-)-L (p < 0.01). For the MAS receptor there were differences between the WT-NL/WT-L (p < 0.001) and AT2(-)-NL/AT2(- )-L (p < 0.001) groups, and also in relation to the WT-L group /AT1(-)-L (p < 0.001) and AT1(-)-L/AT2(-)-L (p < 0.001). For the expression of ACE and ACE2 peptides, there was a statistical difference only for ACE between the types of WT-NL/WT-L groups (p < 0.001). Conclusion: The animals in the AT1(-) group showed less inflammation than the other diseased lines, as well as a lower expression of the Mas and Ang 1-7 receptor. Furthermore, animals from the WT and AT2(-) groups demonstrated similar results in several analyses, showing that the blockade of the AT1 receptor, on molecular effects, is more positive (AU).

Identifying potential anti-COVID-19 pharmacological components of traditional Chinese medicine Lianhuaqingwen capsule based on human exposure and ACE2 biochromatography screening

Lianhuaqingwen (LHQW) capsule, a herb medicine product, has been clinically proved to be effective in coronavirus disease 2019 (COVID-19) pneumonia treatment. However, human exposure to LHQW components and their pharmacological effects remain largely unknown. Hence, this study aimed to determine human exposure to LHQW components and their anti-COVID-19 pharmacological activities. Analysis of LHQW component profiles in human plasma and urine after repeated therapeutic dosing was conducted using a combination of HRMS and an untargeted data-mining approach, leading to detection of 132 LHQW prototype and metabolite components, which were absorbed

Identification of plant defence regulators through transcriptional profiling of Arabidopsis thaliana cdd1 mutant.

A sustainable balance between defence and growth is essential for optimal fitness under pathogen stress. Plants activate immune response at the cost of normal metabolic requirements. Thus, plants that constitutively activate defence are deprived of growth. Arabidopsis thaliana mutant constitutive defence without defect in growth and development1 (cdd1) is an exception. The cdd1 mutant is constitutive for salicylic acid accumulation, signalling, and defence against biotrophic and hemibiotrophic pathogens, without having much impact on growth. Thus, cdd1 offers an ideal genetic background to identify novel regulators of plant defence. Here we report the differential gene expression profile between cdd1 and wild-type plants as obtained by microarray hybridization. Expression of several defence-related genes also supports constitutive activation of defence in cdd1. We screened T-DNA insertion mutant lines of selected genes, for resistance against virulent bacterial pathogen Pseudomonas syringae pv. tomato DC3000 (Pst DC3000). Through bacterial resistance, callose deposition and pathogenesis-associated expression analyses, we identified four novel regulators of plant defence. Resistance levels in the mutants suggest that At2g19810 and [rom] At5g05790 are positive regulators, whereas At1g61370 and At3g42790 are negative regulators of plant defence against bacterial pathogens.

Lifestyle intervention might easily improve blood pressure in hypertensive men with the C genotype of angiotensin II type 2 receptor gene

BACKGROUND/OBJECTIVES: Recent studies have reported an association of the angiotensin II type 2 receptor (AT2R) 3123Cytosine/Adenine (3123C/A) polymorphism with essential hypertension and cardiovascular diseases. The purpose of the study was to investigate whether the AT2R 3123C/A polymorphism affects blood pressure for free-living hypertensive men during a 5-month intervention period. SUBJECTS/METHODS: The subjects were free-living hypertensive Japanese men aged 40 to 75 years who agreed to intervention in the period from 2004 to 2011. Detection of the AT2R 3123C/A polymorphism was determined by polymerase chain reaction. The dietary intervention was designed to decrease salt level and to increase potassium level through cooking instructions and self-monitoring of the diet. The exercise session consisted of activities such as stretching, resistance training, and walking. Blood pressure, urinary sodium and potassium excretion, dietary and lifestyle data, and non-fasting venous blood sample were collected at baseline and after the intervention period. RESULTS: Thirty nine subjects were eligible for participation and the follow-up rate was 97.4%. The C allele proportion was 57.9%. AT2R 3123C/A polymorphism was X-chromosome-linked, therefore we analyzed the C and A genotypes. At baseline, no significant differences were observed between the genotype groups. After the intervention, there were no significant differences in lifestyle habit between the groups. Nevertheless, the estimated salt excretion (g/day) was significantly decreased only in the C genotype (13.0-10.3, P = 0.031). No significant change was observed in systolic blood pressure (SBP) (mmHg) in the A genotype, but a significant decrease was observed in the C genotype (150.0-141.5, P = 0.024). CONCLUSTIONS: In the C genotype, it might be easy to improve SBP through lifestyle intervention in free-living hypertensive Japanese men, however generalization could not be achieved by the small sample size.

Cross Talk between Oxidative Stress, PPAR- γ and RAS System in Diabetic Rat Heart Exposed to Ischemia / Reperfusion Injury.

The present study aimed at clarifying the cross talk between peroxisome proliferator-activated receptor-gamma (PPAR- γ), cardiac reactive Oxygen species (ROS) and Renin-Angiotensin System (RAS). Methods: A total of 90 male albino rats were used. The rats were divided into: Group 1: Control group, Group 2: Type 2 diabetic rats, Group 3: PPARγ agonist protected type 2 diabetic rats. Group 4: Antioxidant protected type2 diabetic rats, Group 5: Metformin treated type 2 diabetic rats. Blood samples were collected for measurement of FBS and fasting insulin. Half the number of each group was sacrificed and the heart excised and perfused, from the rest of the group small piece from the heart was taken for estimation of malondialdehyde (MDA), Angiotensin 2 Receptor (AT2R) and Angiotensin Converting Enzyme 2 (ACE2) gene expression. Results: Treatment with pioglitazone and Vitamin E significantly lowered blood glucose, insulin levels and Homeostasis Model Assessment Insulin Resistance (HOMA IR). However, values did not return to control values. Pioglitazone and Vitamin E improved myocardial performance and percentage recovery following ischemia reperfusion. The cardioprotective effect was more pronounced in the pioglitazone group. This positively correlated with decreased MDA levels and increased AT2R and ACE2 expression in cardiac tissue. Conclusion: Pioglitazone and Vitamin E in type 2 DM significantly offered cardioprotection through improving the diabetic condition and / or decreasing MDA levels.

Relationship between the polymorphism of the AT2R gene 1675A/G and the effect of indapamide sustained release tablets in female hypertension patients / 中国综合临床

Objective To investigate the relationship of the1675A/G polymorphism of AT2 gene with the therapeutic effect of indapamide sustained release tablets in female patients with primary hypertension.Methods Two hundred and twenty female patients with primary hypertension were treated with Indapamide Sustained Release Tablets ( 1.5 mg · qd) for 8 weeks.The blood samples from the patients were collect to determine AT2 gene polymorphism by PCR combined with HRM and sequencing.Results Two hundred and five patients completed the test.In female patients,the therapeutic efficacy of indapamide sustained telease tablets among different AT2R genotypes( AA:70.6％,AG:71.6％,GG:71.4％ ) showed no significant difference ( x2=2.53,P=0.49 ),neither do the decline of BP after therapy ( F=0.39 and 0.19 respecrively,P ＞ 0.05).Conclusion The AT2 genotype was assumed to be not correlated to the blood pressure lowering response to Indapamide Sustained Release Tablets in female primary hypertension patients.

Construction of AT2R gene recombinant adenovirus by an efficent method / 医学研究生学报

Objective:To construct the recombinant adenovirus of AT2R by using the method of homogenous recombination in bacteria. Methods:AT2R gene was get from the vector of PUHD-AT2R by PCR ,and subcloned into shuttle vector of pAdTrack-CMV, forming transfer vector of pAdTrack-CMV/AT2R.Then it was linearized with PmeⅠ and transformed into Adeasier-1 cell. The DNA of identified recombinant plasmid was digested with PacⅠ and transfected to 293cells to package adenovirus. The PCR technique was used to detect target gene . The titre of the Ad-AT2R was measured with the aid of GFP expression. Results:PCR test indicated each the recombinant adenovirus contained the insert of AT2R. The titre of purified recombinant adenovirus was 1.5?10 12pfu/ml. Conclusion:The method of homologous recombination in bacteria is more convenient and efficient compared with that in cell .The prepared Ad-AT2R paves a sound foundation for further study.

In Vitro Intrinsic Radiosensitivity Of Human Squamous Cell Carcinoma in Primary Culture / 대한치료방사선과학회지

There are a number of reports suggesting that there may be a correlation between the clinical response to radiotherapy in various tumors and the clonogenic survival of cell lines derived from these tumor following exposure to 2 Gy (SF2). Authors conducted this study to determine SF2 for cells in primary culture from surgical specimens. The tumor tissues with squamous cell carcinoma of uterine cervix and head and neck were obtained. The tumor tissues were disaggregated to single cells by incubating with collagenase typeIV for 2 hours with constant stirring. Single cell suspensions were inoculated in four 24-well plates precoated with cell adhesive matrix. After 24 hours of incubation at 37degree C, rows of four wells were then irradiated, consisting of control set and five other sets each receiving doses of 1,2,3,4, and 6 Gy. After incubation for a total of 13 days, the cultures were stained with crystal violet determine whether cell growth was of epithelial origin, immunocytochemical staining with a mixture of cytokeratin and epithelial monoclonal antibodies were performed on cell cultures. During the period of this study, we received 5 squamous cell carcinoma specimens of head and neck and 20 of uterine cervical carcinoma. Of these, 15 yielded enough cells for radiosensitivity testing. This resulted an overall success rate of 60%. The mean SF2 value for 15 tumors was 0.55+/-0.17 ranging from 0.20 to 0.79. These results indicate that there is a broad range of sensitivities to radiation in same histologic type. So with a large patient population, we plan to determine whether a different SF2 value is associated with tumors that are controlled with radiotherapy than those that are not.

AT2R gene transfer to rat carotid arteries inhibits neointimal hyperplasia after balloon angioplasty / 中国病理生理杂志

0.05).pAdCMV/AT2R transfection significantly decreased the expression of PCNA in neointima at day 14 [(27.29?5.81)% vs(72.25?4.47)%,(68.43?9.12)%,P

Expression of AngⅡ receptors in excess load-pressured hypertrophied cardiac muscles of rats / 西安交通大学学报(医学版)

Objetive To study the effects of captopri on AT1 and AT2 receptors of load-pressured hypertrophied cardiac muscles of adult rats.Methods Using the methods of narrowing and contraction of the aorta of adult healthy rats to establish the model of animals with hypertrophied cardiac and to observe the changes of receptors AT1 and AT2 of hypertrophied cardiac muscles of AC rats by comprehensive applications of cardiac-tube,immunity tissue chemistry and the technique of image disjunction.Results Left ventricular and cardiac muscles of rats were increased remarkably as the pressure-loading time lasted.After using captopri,left ventricular hypertrophied one was decreased more significantly than that in control group.The expression of receptor AT 1 was increased remarkably as the pressure-loading time lasted.The expression of receptor AT2 was increased transiently,after that reached about as high as control group.AT1/AT2 was increased one week after operation,and then was progressively decreased.After AC,captopri prevented hypertrophied cardiac muscles and receptor AT1 in cardiac muscles.AT1/AT2 was remarkably decreased.Conclusion Load-pressured cardiac hypertrophy may increase the expression of AT1.The expression of receptor AT 2 was increased transiently.The main working mechanism of captopri's preventing and treating the load-pressured cardiac hypertrophy in adult rats may be lowering the expression of AT1 and may effectively block the cardiac hypertrophy regulation precess of Ang ll via AT1 preventing load-pressured cardiac hypertrophy in adult rats.