Bioactive Ingredients in K. pinnata Extract and Synergistic Effects of Combined K. pinnata and Metformin Preparations on Antioxidant Activities in Diabetic and Non-Diabetic Skeletal Muscle Cells.

With healthcare costs rising, many affected by ailments are turning to alternative medicine for treatment. More people are choosing to complement their pharmacological regimen with dietary supplements from natural products. In this study, the compound composition of Kalanchoe Pinnata (K. pinnata) and the effects of combined preparations of K. pinnata and metformin on antioxidant activity in human skeletal muscle myoblasts (HSMMs) and human diabetic skeletal muscle myoblasts (DHSMMs) were investigated. Ultraperformance liquid chromatography fusion orbitrap mass spectrometry (UPLC-OT-FTMS) identified biologically active flavanols in K. pinnata. The main compounds identified in locally grown K. pinnata were quercetin, kaempferol, apigenin, epigallocatechin gallate (EGCG), and avicularin. Antioxidant results indicated that a combinatorial preparation of K. pinnata with metformin may modulate antioxidant responses by increasing the enzymatic activity of superoxide dismutase and increasing levels of reduced glutathione. A combination of 50 µM and 150 µg/mL of metformin and K. pinnata, respectively, resulted in a significant increase in reduced glutathione levels in non-diabetic and diabetic human skeletal muscle myoblasts and H2O2-stress-induced human skeletal muscle myoblasts. Additionally, a K. pinnata treatment (400 µg/mL) alone significantly increased catalase (CAT) activity for non-diabetic and diabetic human skeletal muscle myoblasts and a H2O2-stress-induced human skeletal muscle myoblast cell line, while significantly lowering malondialdehyde (MDA) levels. However, the treatment options were more effective at promoting cell viability after 24 h versus 72 h and did not promote cell viability after 72 h in H2O2-stress-induced HSMM cells. These treatment options show promise for treating oxidative-stress-mediated pathophysiological complications associated with type II diabetes.

The Effect of Acute and Chronic Thermotherapy on Type 2 Diabetic Skeletal Muscle Gene Expression and Inflammatory Markers.

BACKGROUND: Type 2 diabetes (T2D) is a chronic illness associated with resistance to or defective insulin secretion. This study investigates the effects of thermotherapy on cell viability, gene expression and inflammation in skeletal muscle cell lines. METHODS: Healthy and T2D human skeletal muscle cell lines (HSMM and D-HSMM, respectively) were subjected to acute or chronic thermo-therapy (AT or CT, respectively). CT consisted of a 30 min exposure to 40 °C, three times a week for three weeks; AT was a one-time exposure. RESULTS: A significant decrease in D-HSMM cell viability percentage followed AT; however, no significant change occurred in CT. HSMM yielded the highest elevations of genes following CT. In D-HSMM, both treatments yielded gene upregulation. Both treatments significantly down-regulated IL-1ß, IL-6, IL-10 and TNF-α in HSMM. AT significantly decreased IL-1ß, IL-6 and upregulated IL-10 and TNF-α levels in D-HSMM, while CT yielded a reduction in IL-4, TNF-α and an upregulation of IL-6 and IL-10. CONCLUSIONS: An increase in gene expression indicates actin activity and cellular responses, suggesting an increase in transcriptional regulation. The upregulation of IL-6 and IL-10 in D-HSMM negatively correlated with a decrease in TNF-α and IL-1ß, indicating improved adverse inflammatory effects associated with the disease.

Thermotherapy Effects on Healthy and Type 2 Diabetes Human Skeletal Muscle Myoblast Cell Lines.

Diabetes mellitus is a chronic metabolic disease characterized by elevated blood glucose levels with associated disordered carbohydrate and lipid metabolism. Type 2 diabetes (T2D) specifically has been shown to cause a decrease in skeletal muscle mass due to oxidative stress. This study investigated a treatment option for T2D through thermotherapy on healthy (HSMM) and T2D (D-HSMM) human skeletal muscle cells. The goals were to determine the effects of thermotherapy, long-term (chronic) and short-term (acute), on HSMM and D-HSMM cell viabilities and oxidative stress. HSMM and D-HSMM cells were grown to confluency, harvested, and counted to determine density. Acute and chronic heat treatments were applied to both cell lines. The chronic treatment consisted of a 30-minute exposure to 40°C, three times a week for three weeks; the acute treatment was a one-time exposure. Oxidative stress assays and cell viabilities were tested 24 hours after heat treatments. Results indicated no significant effect on the cell viability of HSMM and D-HSMM cells. The acute treatment had a significant increase (p ≤ 0.05) of MDA concentration compared to the chronic treatment. The chronic treatment had a significant increase (p ≤ 0.05) in catalase activity compared to the acute treatment. The SOD activity had no significant change (p > 0.05) between the chronic and acute treatments. In conclusion, acute thermotherapy may not be beneficial for skeletal muscle cells due to the observed increase in oxidative stress, especially in the D-HSMM cells.

Effect of Combined K. pinnata and Metformin Preparation on Inflammatory Cytokines in Normal and Diabetic Skeletal Muscle Cells.

Diabetes Mellitus is associated with systemic inflammation and oxidative stress, which may play a central role in the development of diabetic complications. In this study, combined preparations of Kalanchoe pinnata and metformin were investigated to determine the effects on inflammatory activity in human skeletal muscle myoblasts (HSMMs) and human diabetic skeletal muscle myoblasts (DHSMMs). Results showed that combinatorial preparations sustained cell viability for 3 days in both HSMM and DHSMM cells. However, a significant decrease in cellular viability occurred for both cell lines on day 5. Results also indicate that combinatorial preparations of K. pinnata may modulate immune responses by significantly upregulating proinflammatory markers, interleukin (IL) 2, and tumor necrosis factor-alpha, and upregulating the anti-inflammatory marker, IL-10, in HSMM and DHSMM cells. The combined preparations significantly downregulated the anti-inflammatory glycoprotein IL-6 in both diabetic and nondiabetic human skeletal muscle cells. The findings suggest that combined preparations of K. pinnata and metformin might be a potential immune-modulating agent that may promote inflammation and adversely affect the outcome of diabetic patients.

Combined Inositol Hexakisphosphate and Inositol Supplement Consumption Improves Serum Alpha-Amylase Activity and Hematological Parameters in Streptozotocin-Induced Type 2 Diabetic Rats.

This study evaluated the effect of combined inositol hexakisphosphate (IP6) and inositol supplement on organ weight, intestinal ATPase activities, complete blood count, and serum analytes in streptozotocin (STZ)-induced type 2 diabetic rats. High-fat diet and a single intraperitoneal injection of streptozotocin (35 mg/kg body weight) were used to induce type 2 diabetes mellitus in Sprague-Dawley rats. The diabetic groups were then treated with either combined IP6 and inositol supplement or glibenclamide for four weeks. Organ weights, intestinal ATPase activities, complete blood count, serum α-amylase, total protein, albumin, and globulin content were determined. Pancreatic weight was significantly reduced while relative kidney and liver weights were elevated in the group treated with combined IP6 and inositol supplement compared to the nondiabetic control. Serum α-amylase activity for the glibenclamide and combination treated groups was significantly improved compared to that of the untreated diabetic group. Red cell distribution width percentage was significantly lower in the combination treated group compared to that in the untreated diabetic group, while intestinal ATPase activities were unaffected by the treatment regime. Combined IP6 and inositol supplement consumption may protect people with diabetes from increased risk of cardiovascular diseases due to the supplement's ability to maintain red cell distribution width percentage towards the normal control group.

Effects of Ligusticum porteri (Osha) Root Extract on Human Promyelocytic Leukemia Cells.

BACKGROUND: Ligusticum porteri roots have been traditionally used in folk medicine, but the scientific basis is unclear. OBJECTIVE: To investigate the cytotoxicity, antioxidant, and immunomodulatory effects of L. porteri root extract on human promyelocytic leukemia (HL-60) cells and H2O2-induced oxidative damaged HL-60 cells. MATERIALS AND METHODS: HL-60 cells were incubated with different concentrations of root extract, and cells were harvested for viability assays on day 3 and 7. Cytokine levels (interferon-gamma [IFN-γ], interleukin-2 [IL-2], and interleukin-10 [IL-10]) and antioxidant indexes (malondialdehyde [MDA], reduced glutathione [GSH], superoxide dismutase [SOD], and catalase [CAT]) in H2O2-induced-stressed HL-60 were measured after 2 days. RESULTS: The viability of HL-60 challenged with H2O2 declined by 42% compared to unstressed cells. After 7 days of incubation with 200 or 400 µg/mL L. porteri, the viability of HL-60 cells was two-fold higher than the control. Stressed HL-60 cells treated with 100, 200, and 400 µg/mL L. porteri reduced the lipid peroxidation by 12%-13%. We noted an increase in GSH levels, SOD and CAT activities in stressed HL-60 supplemented with 400 µg/mL root extract. Treatment with 400 µg/mL L. porteri significantly (P < 0.05) increased IFN-γ and IL-2 in H2O2-challenged cells. CONCLUSION: Our data do not support the use of the extract as an antiproliferation and differentiation therapy for acute promyelocytic leukemia. The protective function of L. porteri root extract against oxidative stress could occur through increasing GSH and higher expression of antioxidant enzymes. SUMMARY: Findings from this study may not support the use of Ligusticum porteri root extract as an antiproliferation and differentiation therapy for acute promyelocytic leukemiaOur data suggest that L. porteri root extract may be a potential antioxidant with protective effect against the oxidation of reduced glutathione (GSH)Treatment with L. porteri root extract may be effective in preventing oxidative damage through increasing the activities of antioxidant enzymes (superoxide dismutase [SOD] and catalase [CAT]) in acute promyelocytic leukemia cells.

Investigation of the cytotoxicity, antioxidative and immune-modulatory effects of Ligusticum porteri (Osha) root extract on human peripheral blood lymphocytes.

OBJECTIVE: Ligusticum porteri is a traditional Native American herb. The roots of L. porteri are traditionally used in the treatment of many diseases, however, its cytotoxicity, antioxidative and immune-modulatory effects need to be investigated. In this study, we evaluated the effects of the root extract at different doses on human peripheral blood lymphocytes (PBLs). METHODS: The lymphocytes were incubated with different concentrations of the root extracts (0, 50, 100, 200, and 400 µg/mL) and harvested every 6 h for 2 d (P<0.05). The protective effect of the herb against oxidative damage was determined by inducing oxidative stress with the administration of 50 µmol/L of hydrogen peroxide (H2O2). RESULTS: Treatments with L. porteri at 200 and 400 µg/mL increased the viability of PBLs. The deleterious effect of H2O2 was ameliorated by 400 µg/mL L. porteri treatment. Addition of 400 µg/mL L. porteri reduced lipid peroxidation in stressed PBLs by 94% (P<0.05). Treatment with 400 µg/mL of L. porteri resulted in a 26.4% increase of reduced glutathione levels. Activities of superoxide dismutase and catalase increased by 17.5% and 55.2% respectively, when stressed PBLs were treated with 400 µg/mL L. porteri for 2 d (P<0.05). Treatment with 400 µg/mL L. porteri increased interferon-γ and interleukin-2 expressions in H2O2-challenged PBLs (P<0.05), however, the root extract did not cause a significant difference in interleukin-10 levels compared to the control (P>0.05). CONCLUSION: The findings suggest that L. porteri might be a potential immune-modulating agent involving protective effects against oxidative damage.

Oxidative Stress Parameters and Erythrocyte Membrane Adenosine Triphosphatase Activities in Streptozotocin-induced Diabetic Rats Administered Aqueous Preparation of Kalanchoe Pinnata Leaves.

BACKGROUND: Diabetes mellitus is a chronic metabolic disease that according to the World Health Organization affects more than 382 million people. The rise in diabetes mellitus coupled with the lack of an effective treatment has led many to investigate medicinal plants to identify a viable alternative. OBJECTIVE: To evaluate red blood cell (RBC) membrane adenosine triphosphatase (ATPase) activities and antioxidant levels in streptozotocin-induced diabetic rats administered aqueous preparation of Kalanchoe pinnata leaves. MATERIALS AND METHODS: Diabetes mellitus was induced in rats by a single administration of streptozotocin (60 mg/kg). Diabetic rats were then treated with aqueous K. pinnata preparation (three mature leaves ~ 9.96 g/70 kg body weight or about 0.14 g/kg body weight/day) for 30 days. Serum glucose, RBC membrane ATPase activities, and antioxidant levels were determined. RESULTS: We noted weight loss and reduced food consumption in the treated diabetic group. Serum glucose levels were reduced in the treated diabetic group compared to the other groups. Superoxide dismutase activity and glutathione levels were not significantly elevated in the treated group compared to the diabetic group. However, serum catalase activity was significantly (P < 0.05) increased in the treated diabetic group compared to the other groups. Serum thiobarbituric acid reactive substances were not significantly altered among the groups. There was a significant (P < 0.05) increase in Mg(2+) ATPase activity and a nonsignificant increase in Na(+)/K(+) ATPase activity in the RBC membrane of the treated diabetic group compared to the diabetic group. CONCLUSION: The consumption of aqueous preparation of K. pinnata may accrue benefits in the management of diabetes by lowering oxidative stress often associated with the disease and improving the availability of cellular magnesium through an increase in the magnesium ATPase pump in the RBC membrane for increased cellular metabolism of glucose through the glycolytic pathway. SUMMARY: We noted weight loss and reduced food consumption in the diabetic rats treated with K. pinnata preparationSerum glucose levels were reduced in diabetic rats treated with K. pinnata preparationSerum catalase activity was significantly (P < 0.05) increased in diabetic rats treated with K. pinnata preparationWe also noted a significant (P < 0.05) increase in Mg(2+) ATPase activity in the RBC membranes of diabetic rats treated with K. pinnata preparationOverall, the consumption of aqueous preparation of K. pinnata lowered oxidative stress often associated with diabetes and improved availability of cellular magnesium through an increase in magnesium ATPase pump in the RBC membrane.