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Effect of cilnidipine as an antihypertensive agent on two forms (L-NAME and L-NAME+4%NaCl) of hypertension in rats.

2022-10-01T00:00:00Z

Title: Effect of cilnidipine as an antihypertensive agent on two forms (L-NAME and L-NAME+4%NaCl) of hypertension in rats. Authors: SHAIKH, GOUHER BANU Abstract: Background: A calcium channel blocker (CCB) of the fourth generation, cilnidipine, is dihydropyridine. It is inhibiting calcium channels of both the L- and N-type. Vascular smooth muscle contains L-type calcium channels, while presynaptic nerve terminals contain N-type calcium channels. Vasodilating effects from cilnidipine are slow-acting and persistent. Very few research studies have been done to clarify how N-type calcium channel blockers affect hypertensive rats whose nitric oxide production has been suppressed. Aim and objectives: The purpose of our study was to show protective effect of cilnidipine on L-NAME or L-NAME plus salt induced experimental hypertension rats. Objectives of our study are to assess cardiovascular hemodynamic parameters like heart rate, mean arterial blood pressure, symapathovagal balance by heart rate variability analysis (HRV), systemic and renal oxidative stress (serum and kidney tissue MDA), renin angiotensin system activity (Kidney and serum VEGF, NOS3 and ACE protein expression and serum and urinary Ang II level) in N G -nitro-L-arginine methyl ester hydrochloride (L-NAME) induced hypertensive rat model. Further we investigated (24hour protein excretion, Creatinine clearance, Renal fibrosis/glomerulosclerosis) as markers of renal injury in response to L-NAME and L NAME plus salt induced hypertensive rats with or without cilnidipine treatment. Material and methods: 36 male Albino Wister rats were collected from institutional animal house (six rats in each group). Group1 vehicle treated (control), group2 cilnidipine (2mg/kg body weight/day) treated, group3 treated with L-NAME (40 mg/kg body weight/day), group4 treated with L-NAME and cilnidipine, group 5 treated with L-NAME and 4% sodium chloride (4% NaCl), group 6 L-NAME, cilnidipine and salt treated. All experimental animals underwent gravimetry and after28-day percentage of body weight gain calculation was made. Estimates were made for haematological variables such as Haemoglobin (g/dl), RBC count (million cells/mm3), and haematocrit (percentage). HRV analysis was performed to evaluate changes in the cardiovascular autonomic system, Heart rate (HR) and blood pressure (BP) were monitored every week for 28 days as examples of cardiovascular hemodynamic events. Blood pressure was recorded by non-invasive tail cuff method. Oxidative stress was assessed by estimating serum and kidney tissue MDA levels. Serum and kidney tissue nitric oxide levels were measured as nitrosative stress markers. Proteinuria and creatinine clearance were measured. kidney function parameters (serum urea and creatinine) were also measured. Serum eNOS, Ang II and urinary Ang II levels were quantitatively measured by ELISA technique. Relative expression of serum and kidney tissue NOS3, ACE and VEGF protein levels were done by Western Blotting. Histopathological examination of the Aorta and kidney tissue was done. Results: Lower percentage body weight gain was seen after L-NAME administration, and changes in the sympathovagal balance and sympathetic overactivity were discovered by HRV analysis. Heart rate was lower and mean arterial pressure was higher in Hypertensive rats. There is increase in serum and kidney tissue oxidative stress. Marked decreased in serum and kidney tissue NO and NOS3 levels. There is increased serum and kidney tissue VEGF and ACE protein expression. There is increase in serum and urinary Ang II levels. We also observed proteinuria and decreased creatinine clearance in NO deficient and salt supplemented hypertensive rats. Cilnidipine treatment was able to 1) reduce MAP and Heart rate 2) reduce sympathetic activity 3) decrease serum and kidney tissue oxidative stress 4) increase

Plasminolytic Components And Their Receptors In Pathogenesis Of Preeclampsia

2022-01-01T00:00:00Z

Title: Plasminolytic Components And Their Receptors In Pathogenesis Of Preeclampsia Authors: Ruikar, Komal Abstract: Background Preeclampsia (PE) is a multisystemic pregnancy disorder affecting 2-8% of pregnancies and remains a major cause of maternal and fetal morbidity and mortality. Despite decades of research the underlying cause of preeclampsia is still not clear. The pathophysiology of preeclampsia is complex wherein the placenta plays a central role. The primary pathology appears to be at the maternal fetal interface and is characterized by poor trophoblastic invasion of the uterus. Preeclampsia is associated with failure of endovascular invasion and spiral artery remodelling and plays the central role in pathogenesis of disease. VEGF, a well-known angiogenic factor produced by placental cells, plays a central role in placental pathogenesis of PE. Annexin A2 (ANXA2) is a profibrinolytic receptor required for plasminolysis, which is an important step in the formation of new blood vessel along with VEGF.ANXA2 increases tissue plasminogen activator (tPA) mediated plasmin generation and plasminogen activator inhibitor (PAI-1) inhibit the tPA.Preeclampsia is also associated with maternal, placental aggravated inflammatory response and generalized endothelial damage AnnexinA1 (ANXA1) is glucocorticoid regulated protein regulates a wide range of cellular and molecular steps of the inflammatory response and is implicated in resolution of inflammation.Galectin-3(Gal-3), β-galcotoside-binding lectin participates in many functions, both intra- and extracellular. Recently it has been shown that galectin-3 modulates the inflammation. Role of ANXA1 and Gal-3 is poorly studied in context with human reproductive disease like PE. Therefore, the present study examined the expression of above proteins which are involved in plasminolysis, angiogenesis and modulation of inflammation and their association in the placental bed of pregnancy with and without PEObjectives a. To evaluate the alterations in gross placental morphology of PE compared to placenta of normal women. b. To demonstrate expression pattern ofANXA2, ANXA1, VEGF, tPA, PAI-1, EGFR and Gal-3 in placental tissue from women with and without Preeclampsia. c. To evaluate the correlation between expression of these downstream plasminolytic proteins at the membrane of the placental cellular component to get insight into their possible relationship to placental angiogenesis and inflammation to verify whether it has some role in the development of Preeclampsia. Material and methods The study group comprised of placental tissues procured from gestations with PE (n = 40) and without (n = 30) PE. The expression of ANXA2, ANXA1, VEGF, tPA, PAI-1, EGFR and Gal-3 in the placental villous tissue was evaluated quantitatively by means of IHC, Western blotting and RT-PCR. Results Expression analysis illustrated that significant decrease in the expression of growth proteins VEGF, EGFR and profibrinolytic receptor ANXA2 in PE group and increase expression of tPA and PAI-1 compared with the normotensive control group. Expression of inflammation modulatory proteins ANXA1 and Gal-3 in PE group was more compared with the normotensive control group (P< 0.05) Conclusion Decreased expression of ANXA2 and VEGF with increased expression of PAI-1 is mainly responsible for altered angiogenic and fibrinolytic activity inPE. The increased expression of AnxA1 and Gal-3 in placental bed may be associated with a systemic inflammatory response in PE, suggesting role of above proteins in PE pathogenesis.

Implication of oral contraceptive use to phenotypic expression pattern of receptors in breast cancer

2022-01-01T00:00:00Z

Title: Implication of oral contraceptive use to phenotypic expression pattern of receptors in breast cancer Authors: Khode, Vitthalsa Abstract: Background: Triple negative breast carcinoma (TNBC) is a breast cancer sub-type associated with high mortality rate and inadequate therapeutic options. Clinical data indirectly implicates where Oral Contraceptive Pill (OCP) usage is high, prevalence of Estrogen Receptor+ (ER+) breast cancer is high and prevalence of TNBC is low. This has lead to our hypothesis that OCP use may add to risk of ER+ breast cancer and OCP use may reduce the risk of TNBC. In in-vitro study we tried to differentiate the effect of estrogen on development of ER+ and triple negative breast cancer tumor affecting Epidermal growth factor receptor EGFR expression in respective cancer cell lines as TNBC commonly displays EGFR. It is known that effective EGFR degradation results in suppression of tumor in various models. Aims and Objectives: We aimed at to comparing the prevalence and association of sub-types of breast cancer in OCP users and OCP non-users among woman 30 to 60 years of age, and in-vitro study we aimed at treating MDA-MB-231 cell lines with Cycloheximide with or without 17β-estrdiol to observe whether 17β-estradiol leads to EGFR degradation. We also aimed at whether degradation occurs through ubiquitination pathway. Methods : This hospital-based observational human study of three year duration included 155 subjects of primary invasive breast cancer who got admitted at our institution. The data was obtained for ER, PR, HER2 condition, clinical classification and data related to demographic factors, reproductive history, and history of OCP use. They were divided into two groups. Group-1 included 48 patients with history of OCP use and group-2 included 107 patients who did not use OCP. In in-vitro study MDA- MB-231 cells were treated with 17β-estradiol (E2) and EGFR expression was evaluated by western blotting at different intervals by using Cycloheximide chase. To gauge ubiquitination pathway of degradation of EGFR in the MDA-MB-231 cell line, MG-132 was utilized. Data was analysed using SPSS-20. Results: A significant increase in prevalence of molecular sub-types ER+, Progesterone Receptor+ (PR+) and Luminal B breast cancers in OCP users was observed compared to non-users. There was considerable decrease in the age at the point of admission in ER+ cancer in OCP users (45.3 years) compared to non-users (52.2years). Whereas in OCP users age at the time of admission of Basal (TNBC) cancer patients (53.1 years) was higher when compared to non-users (45.4years). Logistic regression revealed the likelihood of ER+, PR+ and Luminal B in OCP users was 11%,10% and 13% less respectively with 1 year of higher age against the likelihood of TNBC among OCP users was 18% more and 8% less in non-users. In in-vitro study EGFR expression was reduced with β-estradiol treatment in MDA-MB- 231 cell line with Cycloheximide chase. Upon Treatment with MG-132 and E2, EGFR expression did not reduce suggestive of that Estrogen degrades EGFR by ubiquitination pathway. Conclusions: OCP use may be allied with increase in the prevalence of ER+, PR+ and Luminal B breast cancer. On the contrary OCP use is may be related with delay in the progression of the TNBC. In-vitro study conclusion was that estrogen degrades EGFR in MDA-MB-231 cells and this degradation occurs by ubiquitination

Role of nitric oxide synthase 3 (NOS3) gene expression in patients of pre-eclampsia with special reference to cardiovascular and renal pathophysiology

2022-05-01T00:00:00Z

Title: Role of nitric oxide synthase 3 (NOS3) gene expression in patients of pre-eclampsia with special reference to cardiovascular and renal pathophysiology Authors: Herur, Anita Abstract: Background: Pre-eclampsia (PE) is a disorder that complicates pregnancy and is causing a rise in maternal and fetal morbidity and mortality. Oxidative stress affects patients of preeclampsia and is implicated in the pathogenesis. Gene expression of nitric oxide synthase (NOS) is altered and may contribute to cardiac and renal pathophysiology of pre-eclampsia. Aim: The aim of the study was to determine the relationship between the gene expression of nitric oxide synthase and oxidative/nitrosative stress, which thereby could influence the cardiovascular and renal pathophysiology in pre-eclampsia patients. Methods: Preeclampsia patients and normal pregnancy women were included in this case control study. Arterial blood pressure and heart rate were recorded. Frequency and time domain heart rate variability (HRV) analysis was done using the powerlab software. Levels of malondialdehyde (MDA), total antioxidant capacity and nitric oxide (NO) in the serum were estimated. Renal functions were determined by estimating serum proteins, urea, creatinine, uric acid and urine proteins and creatinine estimation. Urine angiotensinogen was also estimated. Urine protein/creatinine ratio and urine angiotensinogen/creatinine ratio were calculated later. Real-time polymerase chain reaction (RT-PCR) was used for nitric oxide synthase 3 (NOS3) and NOS2 gene profiling. Student’s t-test was used for statistical analysis and p value <0.05 was considered statistically significant. Results: A significantly higher (p<0.0001) mean arterial pressure was seen in the PE group. A significant increase was noted in LF/HF ratio, low frequency (LF) component, and a decrease in high frequency (HF) component of the HRV in pre-eclampsia. The time domain parameters, SDNN, SDANN, RMSSD and pNN50%, of HRV in preeclampsia patients, showed a reduction in their levels. Levels o f serum malondialdehyde were increased (p< 0.0001), and on the other hand, the total antioxidant capacity was decreased in the PE group (p= 0.034). A significant decrease in the serum albumin (p=0.004) and an increase in the serum uric acid (p<0.0001) were seen. Urine protein and urine protein: creatinine ratio were significantly higher (p<0.0001). The urinary excretion of angiotensinogen was significantly reduced (p<0.0001). But, the serum levels of NO did not show a statistically significant reduction (p = 0.20). Gene expression profiling in the PE group showed a down regulation of NOS3 and NOS2 by about 8.49 and 51.05 times respectively. Conclusion: Mean arterial pressure increase in preeclampsia may be due to endothelial dysfunction resulting from oxidative stress. Also, parasympathetic withdrawal along with sympathetic overactivity, noted in pre-eclampsia patients, may suggest cardiovascular risk in them, which may be detected early by HRV analysis. These changes may also trigger renal dysfunction. Nitric oxide could have played an important role as the mRNA expression of both NOS3 and NOS2 genes were reduced. However, cross-talk of NO with other vasoactive /biological substances cannot be overlooked