Pancreatic Lipase inhibition assay of various extracts of leaves of Murraya Koenigii in southern areas of Goa

The objective of the study was to assess the lipase inhibitory activities of chloroformic, methanolic and aqueous extracts from the commonly available Murraya koenigii (L.) Spreng leaves(Rutaceae) in southern villages of Goa, for potential use in the treatment of obesity. Extracs of the leaves of this plant were evaluated for lipase inhibitory activity using porcine pancreatic lipase (PPL: triacylglycerol lipase) and p-nitrophenyl butyrate in an in vitro assay. Among the three extracts screened, chloroformic extract exhibited the highest pancreatic lipase inhibitory activity of 53.42%, followed by methanolic extract (51.88%) and aqueous extract (36.42%), respectively. Chloroformic extract has not been screened for its pancreatic lipase inhibition assay. All the Crude extracts of leaves of Murraya koenigii (L.) Spreng leaves (Rutaceae) have potential as pancreatic lipase inhibitory agents. . Chloroformic extract was found to be most effective and hence can be used as a potent anti-obesity agent to combat hyperlipidemia

Pancreatic Lipase inhibition assay of various extracts of leaves of Murraya Koenigii in southern areas of Goa

The objective of the study was to assess the lipase inhibitory activities of chloroformic, methanolic and aqueous extracts from the commonly available Murraya koenigii (L.) Spreng leaves(Rutaceae) in southern villages of Goa, for potential use in the treatment of obesity. Extracs of the leaves of this plant were evaluated for lipase inhibitory activity using porcine pancreatic lipase (PPL: triacylglycerol lipase) and p-nitrophenyl butyrate in an in vitro assay. Among the three extracts screened, chloroformic extract exhibited the highest pancreatic lipase inhibitory activity of 53.42%, followed by methanolic extract (51.88%) and aqueous extract (36.42%), respectively. Chloroformic extract has not been screened for its pancreatic lipase inhibition assay. All the Crude extracts of leaves of Murraya koenigii (L.) Spreng leaves (Rutaceae) have potential as pancreatic lipase inhibitory agents. . Chloroformic extract was found to be most effective and hence can be used as a potent anti-obesity agent to combat hyperlipidemia

CD45^intCD11b^int MPCs express distinct cytokines.

<p>Intracellular cytokine level in each MPC population was measured by flow cytometry. (a, c) Representative histograms show the expression level of TNF-α and IL-10 in each MPC population after LPS stimulation (left) or after ischemic injury (right). Yellow plot indicates the cytokine production of unstimulated MPCs from normal mice, being used as a control. Red plot and blue plot represent the cytokine expression by CD45^intCD11b^int MPCs and CD45^highCD11b⁺ MPCs, respectively. (b, d) Bar graph indicates the proportion of cells producing TNF-α and IL-10 within each MPC population after LPS stimulation (left) or after ischemic injury (right). Data are displayed as means ± SEM (n = 5/group). *<i>P</i> < 0.05; **<i>P</i> < 0.01; ***<i>P</i> < 0.001; ****<i>P</i> < 0.0001; <i>NS</i>, no statistically significant difference between groups.</p

CD45^intCD11b^int MPCs have different phenotype compared to CD45^highCD11b⁺ MPCs.

<p>(left) Representative histograms of indicated markers. Filled plot shows expression level of each markers by CD45^intCD11b^int MPCs and dashed plot shows one from CD45^highCD11b^<b>+</b> cells. (right) Graphs show mean fluorescence intensity (MFI) of each indicated markers. Data are displayed as means ± SEM (n = 3-5/group). *<i>P</i> < 0.05; **<i>P</i> < 0.01; ***<i>P</i> < 0.001; <i>NS</i>, no statistically significant difference between groups.</p

Systemic administration of liposomal clodronate has more pronounced depletive effects on CD45^intCD11b^int population.

<p>(a) Bar graph shows absolute cell numbers in each population after systemic administration of liposomal clodronate or control liposome in normal mouse kidney. The effect of systemic clodronate is more prominent in CD45^intCD11b^int population compared to CD45^highCD11b^<b>+</b> or total CD45^<b>+</b>CD11b^<b>+</b>. (b) Bar graph shows absolute cell numbers of CD45^<b>+</b>CD11b^<b>+</b> cells after systemic administration of liposomal clodronate or control liposome in wild-type mouse spleen. Data are displayed as mean ± SEM (n = 3/group). *<i>P</i> < 0.05; **<i>P</i> < 0.01; <i>NS</i>, no statistically significant difference between groups.</p

CD45^int population are present in normal mouse kidney.

<p>Representative plots show CD45^int cells and CD45^high cells in normal mouse kidneys. ED improves the identification of CD45^int population compared to MD alone. Data are from one of 11 representative experiments with similar results. ED, enzymatic digestion; MD, mechanical digestion.</p

Characterization of kidney CD45^intCD11b^intF4/80⁺MHCII⁺CX3CR1⁺Ly6C^- “intermediate mononuclear phagocytic cells”

<div><p>Kidney immune cells play important roles in pathogenesis of many diseases, including ischemia-reperfusion injury (IRI) and transplant rejection. While studying murine kidney T cells, we serendipitously identified a kidney mononuclear phagocytic cell (MPC) subset characterized by intermediate surface expression of CD45 and CD11b. These CD45^intCD11b^int MPCs were further identified as F4/80^<b>+</b>MHCII^<b>+</b>CX3CR1^<b>+</b>Ly6C^- cells, comprising ~17% of total CD45⁺ cells in normal mouse kidney (<i>P</i> < 0.01) and virtually absent from all other organs examined except the heart. Systemic clodronate treatment had more significant depletive effect on the CD45^intCD11b^int population (77.3%±5.9%, <i>P</i> = 0.03) than on CD45^highCD11b⁺ population (14.8%±16.6%, <i>P</i> = 0.49). In addition, CD45^intCD11b^int MPCs had higher phagocytic function in the normal kidney (35.6%±3.3% vs. 24.1%±2.2%, <i>P</i> = 0.04), but lower phagocytic capacity in post-ischemic kidney (54.9%±1.0% vs. 67.8%±1.9%, <i>P</i> < 0.01) compared to the CD45^highCD11b⁺ population. Moreover, the CD45^intCD11b^int population had higher intracellular production of the pro-inflammatory tumor necrosis factor (TNF)-α (58.4%±5.2% vs. 27.3%±0.9%, <i>P</i> < 0.001) after lipopolysaccharide (LPS) stimulation and lower production of the anti-inflammatory interleukin (IL)-10 (7.2%±1.3% vs. 14.9%±2.2%, <i>P</i> = 0.02) following kidney IRI, suggesting a functional role under inflammatory conditions. The CD45^intCD11b^int cells increased early after IRI, and then abruptly decreased 48h later, whereas CD45^highCD11b⁺ cells steadily increased after IRI before declining at 72h (<i>P</i> = 0.03). We also identified the CD45^intCD11b^int MPC subtype in human kidney. We conclude that CD45^intCD11b^int F4/80^<b>+</b>MHCII^<b>+</b>CX3CR1^<b>+</b>Ly6C^-population represent a unique subset of MPCs found in both mouse and human kidneys. Future studies will further characterize their role in kidney health and disease.</p></div

CD45^intCD11b^int cells are distinct mononuclear phagocytic population in normal mouse kidney.

<p>(a) Representative plots show that ED yields a greater proportion of CD45^intCD11b^int cells compared to MD alone in normal mice kidney. Numbers on plots represent the percentage of each population among CD45^<b>+</b> population. (b<b>-</b>d) Bar graph shows absolute cell number of each population isolated by ED or MD method. The use of ED significantly increased the absolute number of the CD45^intCD11b^int population (b) and CD45^highCD11b^<b>+</b> population (c), but there was no significant difference in the total number of KMNCs (CD45^<b>+</b> population) (d). Data are displayed as means ± SEM (n = 3/group). ED, enzymatic digestion; MD, mechanical digestion. *<i>P</i> < 0.05; **<i>P</i> < 0.01; <i>NS</i>, no statistically significant difference between groups.</p

Detection of CD45^intCD11b^int MPCs in human kidneys.

<p>Plots show CD45⁺CD15^- population in “normal” kidney tissue from three different de-identified individuals who underwent partial or total nephrectomies to treat renal cell carcinoma. CD45^intCD11b^int MPCs are marked with solid line and CD45^highCD11b⁺ population with dashed line. Numbers indicate the percentage of each population among CD45⁺ cells.</p