Bhola Nath Paul

Indian Herbal Medicines: Possible Potent Therapeutic Agents for Rheumatoid Arthritis

Rheumatoid arthritis (RA) is an autoimmune disease of unknown etiology and is mainly characterized by the progressive erosion of cartilage leading to chronic polyarthritis and joint distortion. Although the exact pathogenesis of the disease has yet not been elucidated, however, studies suggest that cellular proliferation of synoviocytes result in pannus formation which damages the cartilage and bone. Recent reports also support the role of free radicals in its pathogenesis. Apart from the conventional treatment strategies using nonsteroidal anti-inflammatory drugs, disease modifying antirheumatic drugs and glucocorticoids, newer and safer drugs are continuously being searched, as long term usage of these drugs have resulted in adverse effects. Alternative medicine provides another approach for treatment of RA and currently a number of medicinal plants are under scientific evaluation to develop a novel drug. There is a dire need to investigate the complete therapeutic potential and adverse effects, if any, of these herbals for providing newer and safer treatment options with minimum side effects. In this review we have tried to explore various Indian ancient Ayurvedic, Unani and Tibbi, as also some Chinese and Korean, herbals for their potential to treat RA.

Swertia chirayita Mediated Modulation of Interleukin‐1β Interleukin‐6, Interleukin‐10, Interferon‐γ, and Tumor Necrosis Factor‐α in Arthritic Mice

Abstract The effect of aqueous extract of Swertia chirayita stem on the pro‐ and anti‐inflammatory cytokines balance in primary joint synovium of adjuvant‐induced arthritic mice has been studied. The level of pro‐inflammatory cytokines was found elevated in the joint synovium of arthritic mice in comparison to normal joints. Administration of S. chirayita extract in varying doses through the oral route did not modulate the proinflammatory cytokines on day 2. In contrast, by day 12, a dose dependent (0, 11.86 and 23.72 mg/kg body weight) reduction of tumor necrosis factor‐α (INF‐α) interleukin‐1β, (IL‐β) and interferon‐γ, (IFN‐γ) and elevation of Interleukin‐ 10 (IL‐10) was observed in the joint homogenates of arthritic mice. Interleukin‐6 (IL‐6) was not down regulated in joint homogenate of arthritic mice at the dose 11.86 mg/kg but at higher doses (23.72 and 35.58 mg/kg) significant reduction was observed. The aqueous extract was found to possess two polar compounds, amerogentin and mangiferin but was devoid of swerchirin, chiratol, methyl swetianin, and swertanone. Mangiferin has been reported to possess potent anti‐inflammatory property and we presume its presence in the aqueous extract of S. chirayita is responsible for reducing TNF‐α, IL‐1β, IL‐6, and IFN‐γ and/or elevating IL‐10 in the joint homogenates of arthritic mice on day 12. This study will help in our understanding of the mechanism of anti‐inflammatory action of S. chirayita in the light of proinflammatory and anti‐inflammatory cytokine balance.

Over expression of resistin in adipose tissue of the obese induces insulin resistance

AIM To compare resistin mRNA expression in subcutaneous adipose tissue (SAT) and its correlation with insulin resistance (IR) in postmenopausal obese women. METHODS A total of 68 postmenopausal women (non obese = 34 and obese = 34) were enrolled for the study. The women of the two groups were age matched (49-70 years). Fasting blood samples were collected at admission and abdominal SAT was obtained during surgery for gall bladder stones or hysterectomy. Physical parameters [age, height, weight, body mass index (BMI)] were measured. Biochemical (plasma insulin and plasma glucose) parameters were estimated by enzymatic methods. RNA was isolated by the Trizol method. SAT resistin mRNA expression was done by real time- reverse transcription polymerase chain reaction (RT-PCR) by using Quanti Tect SYBR Green RT-PCR master mix. Data was analyzed using independent Students t test, correlation and simple linear regression analysis. RESULTS The mean weight (52.81 ± 8.04 kg vs 79.56 ± 9.91 kg; P < 0.001), BMI (20.23 ± 3.05 kg/m(2)vs 32.19 ± 4.86 kg/m(2); P < 0.001), insulin (8.47 ± 3.24 μU/mL vs 14.67 ± 2.18 μU/mL; P < 0.001), glucose (97.44 ± 11.31 mg/dL vs 109.67 ± 8.02 mg/dL; P < 0.001) and homeostasis model assessment index (2.01 ± 0.73 vs 3.96 ± 0.61; P < 0.001) were significantly higher in postmenopausal obese women compared to postmenopausal non obese women. The mean serum resistin level was also significantly higher in postmenopausal obese women compared to postmenopausal non obese women (9.05 ± 5.15 vs 13.92 ± 6.32, P < 0.001). Furthermore, the mean SAT resistin mRNA expression was also significantly (0.023 ± 0.008 vs 0.036 ± 0.009; P < 0.001) higher and over expressed 1.62 fold (up-regulated) in postmenopausal obese women compared to postmenopausal non obese women. In postmenopausal obese women, the relative SAT resistin mRNA expression showed positive (direct) and significant correlation with BMI (r = 0.78, P < 0.001) and serum resistin (r = 0.76, P < 0.001). Furthermore, the SAT resistin mRNA expression in postmenopausal obese women also showed significant and direct association (r = 0.45, P < 0.01) with IR, while in postmenopausal non obese women it did not show any association (r = -0.04, P > 0.05). CONCLUSION Increased SAT resistin mRNA expression probably leads to inducing insulin resistance and thus may be associated with obesity-related disorders in postmenopausal obese women.

GSTM1, GSTT1, and GSTP1 polymorphism in north Indian population and its influence on the hydroquinone-induced in vitro genotoxicity.

Glutathione S transferase (GST) gene polymorphism examined among north Indians and correlated with hydroquinone (HQ) genotoxicity to help in clinical prediction of susceptibility of HQ toxicity. Lymphocytes of individuals with/without GSTM1, GSTT1, and GSTP1 (ile/ile or val/val) were exposed to HQ (20, 40, or 80 μM) and examined chromosomal aberrations (CA) or cytokinesis-block micronucleus assays. Among north Indians the frequencies of GSTM1 (null), GSTT1 (null), and both null were found to be 41.1, 21.9, and 12.7%, whereas frequencies of GSTP1 with (ile/ile) or (ile/val), or (val/val) were 52, 42.1, or 5.9%, respectively. Individuals with null GSTM1, GSTT1, and GSTP1 (val/val) showed inhibition of mitotic index (MI) and significant (p < 0.01) induction of CA as compared to individuals with GSTM1, GSTT1, and GSTP1 (ile/ile). Micronucleus formation was found to be significant (p < 0.05 or 0.01) in both the genotypes. Results indicate that GSTM1, GSTT1 (null), and GSTP1 (val/val) are sensitive to HQ genotoxicity.

Development of novel topical drug delivery system containing cisplatin and imiquimod for dual therapy in cutaneous epithelial malignancy

Abstract Context: Strategy of dual therapy has been proposed to minimize the amount of each drug and to achieve the synergistic effect for cancer therapies. Objective: The aim of this study was to develop an effective drug delivery system for the simultaneous topical delivery of two anti-tumor agents, cisplatin and imiquimod. Material and methods: The preformulation studies were carried out in terms of tests for identification, solubility profile, determination of partition coefficient and simultaneous estimation of both drugs by UV–Visible spectrophotometer and High Performance Liquid Chromatography (HPLC). Drug–drug and drug-excipients interactions were examined by thin layer chromatography, infrared spectroscopy, differential scanning calorimetry (DSC) and X-ray diffraction (XRD). Provesicular drug delivery system (protransfersome gel formulation) have been prepared and characterized by in vitro and in vivo parameters. Results: The mean size, poly dispersity index (PDI) and zeta potential of transfersomal vesicles formed by protransfersome hydration were 429.5 nm, 0.631 and −68.1 Mv, respectively. The prepared formulation showed toxicity on cutaneous squamous cell carcinoma cell line (A-431) at 200 µg (cisplatin) and 1 mg (imiquimod) concentration of drug in combination against control. The cisplatin- and imiquimod-loaded provesicular dual–drug delivery system achieved an optimal antitumor effect, increase in lifespan, antiviral, and toxicity reduction, revealing the advantage of site specific drug delivery and the modified combination therapy. Discussion: Cisplatin delivery through protransfersome gel in combination with imiquimod may potentiate the activity against solid tumors of epidermal origin. Conclusion: Data revealed that combination therapy considerably enhances antitumor efficacy of the drug for skin-cited malignancies.

A novel triazine-aryl-bis-indole derivative inhibits both phosphodiesterase IV and expression of cell adhesion molecules

Asthma, like many inflammation related disorders, has a complex etiology. Drugs targeting multiple pathways may prove more efficacious in these complex disorders. Cyclic 3′,5′-adenosine monophosphate (cAMP) phosphodiesterase IV (PDE IV) is one of the validated targets in bronchial asthma and despite availability of some therapeutic molecules targeting PDE IV, molecules with better properties are desired. Eosinophil/neutrophil infiltration into lung may also be an important component of bronchial asthma in which increased expression of epithelial cell adhesion molecules may play an important role. This study describes the synthesis of a novel class of compounds ‘triazine-aryl-bis-indoles’ having a catechol derived structure constituting a part of ‘triazine’ and a part of ‘bis-indole’ moiety on it. This class of molecules potently inhibited both phosphodiesterase IV and expression of cell adhesion molecules ICAM-1 and VCAM-1. The best molecule of this class (compound 11) inhibited PDE IV activity in vitro, with an IC50 value of 14 μM compared to 12.7 μM for an existing drug rolipram. The compound 11 not only stabilized the cAMP level in human lung epithelial cells (L132) following stimulation with forskolin, but also inhibited TNF-α induced expression of cell adhesion molecules such as ICAM-1 and VCAM-1 in human umbilical vein epithelial cells (HUVECs). It also significantly inhibited the adhesion of human neutrophils to the endothelial monolayer (IC50 = 17.86 μM) in a dose dependent manner. Its absolute bioavailability (in mice) was found to be 70% and its toxicity and pharmacokinetic profiles are excellent. The dual activity of this class of molecules suggests that this class of molecules could have broad therapeutic applications in neutrophil dominant diseases such as severe asthma, COPD and acute lung injury.

IL-6 gene expression in adipose tissue of postmenopausal women and its association with metabolic risk factors

Adipose tissue secretes various kinds of adipokines that controls the glucose and lipid metabolism in humans. The abdominal visceral adipose tissue (VAT) and subcutaneous adipose tissue (SAT) both are associated with metabolic syndrome and insulin resistance. IL-6 is one of the adipokines, which promotes insulin resistance and dyslipidemia in humans. The association of adipokines with metabolic syndrome at protein levels are well documented. However, their association at gene expression level are lacking. The present study was design to investigate IL-6 mRNA expression in adipose tissues (VAT and SAT) and its correlation with metabolic risk factors and insulin resistance (HOMA) in post menopausal women. A total of 108 Asian North Indian post menopausal women, 54 without metabolic syndrome (controls) and 54 with metabolic syndrome (cases) were recruited and evaluated. Overnight fasting blood samples were collected at admission and abdominal visceral and subcutaneous adipose tissues were collected during open abdomen surgery. The results showed significantly (p < 0.05 or p < 0.01 or p < 0.001) higher mean SBP, glucose, insulin, HOMA, TG, VLDL and serum IL-6 while significantly (p < 0.001) lower HDL and estrogen in cases as compared to controls. In cases, the relative mean SAT IL-6 expression was also significantly (p < 0.05) higher as compared to VAT. Further, in cases, the VAT IL-6 expression showed significant (p < 0.05 or p < 0.001) and negative correlation with WC, WHR, glucose, HOMA, TC, LDL and estrogen while SAT IL-6 expression also showed significant (p < 0.05 or p < 0.01 or p < 0.001) and negative correlation with WC, WHR and estrogen. The Cox regression analysis found VAT IL-6 mRNA expression the significant (p < 0.05 or p < 0.01) an independent predictor of WC, HOMA, TC, LDL and estrogen while SAT IL-6 mRNA expression the significant (p < 0.01) an independent predictor of TG and VLDL. The study concluded that IL-6 expressions of both visceral and subcutaneous tissues may be associated with metabolic risk factors in postmenopausal Asian North Indian women.

Adiponectin mRNA in adipose tissue and its association with metabolic risk factors in postmenopausal obese women.

OBJECTIVEThe present study evaluates adiponectin mRNA in visceral adipose tissue (VAT) and subcutaneous adipose tissue (SAT) and also evaluates its association with metabolic risk factors in postmenopausal obese women.DESIGNA case control study was carried out on postmenopausal women (n = 68), in which 34 were obese and 34 were non-obese. Blood sample, visceral and subcutaneous adipose tissues were obtained. Adiponectin mRNA levels were measured by Real Time-RT PCR.RESULTSThe mean (± SEM) serum adiponectin (28.39 ± 2.52 vs. 20.56 ± 1.13), VAT (0.362 ± 0.098 vs. 0.048 ± 0.005) and SAT (0.222 ± 0.035 vs. 0.042 ± 0.007) adiponectin mRNA levels were significantly lower (p<0.001) in obese than non-obese. However, the mean VAT and SAT adiponectin mRNA levels were similar (p>0.05) between the groups. Further, the mean glucose and TG levels were significantly (p<0.01 or p<0.001) higher, while HDL was lower (p<0.01) in obese than non-obese. Furthermore, VAT adiponectin mRNA also showed significant (p<0.05) and inverse association with TG, while direct association with HDL and both the associations were independent of BMI and WC (waist circumference).CONCLUSIONThe BMI and WC independent and significant association of VAT adiponectin mRNA with TG and HDL suggest its potential modulatory role in lipid metabolism in postmenopausal obese women.

Carryover of cigarette smoke effects on hematopoietic cytokines to F1 mouse litters.

Neutrophils have been implicated in the pathogenesis of COPD, being recruited into the lung in response to cigarette smoke (CS) inhalation and responsible for the release of proteases and oxidant-producing enzymes, resulting in bronchitis and emphysema. Several hematopoietic cytokines are involved in neutrophil growth and recruitment; however, little is known about the effects of CS on hematopoietic cytokines are transmitted between generations. In the present investigation we evaluate the expression of hematopoietic and proinflammatory cytokines in different organs of female F(0) mice subjected to sub-chronic CS exposure, and in F(1) litters. Virgin female Balb/c mice inhaled either air or air containing CS for 90 days. The specific resistance of the airways (sRaw) was evaluated and, thereafter, the mice were mated with unexposed adult males. The levels of granulocyte-macrophage colony stimulating factor (GM-CSF), granulocyte-colony stimulating factor (G-CSF), interleukin-6 (IL-6), IL-1β and TNF-α mRNA and protein were evaluated in the bone marrow, amniotic fluid and bronchoalveolar lavage fluid (BALF) of F(0) dams at gestation day(14) (gd(14)) and the bone marrow, BALF and lungs of F(0) dams and F(1) littermates at post natal day(21) (pnd(21)). At gd(14), overexpression of GM-CSF, G-CSF and IL-6 mRNA and protein was observed in the bone marrow, amniotic fluid and BALF of F(0) dams. These hematopoietic cytokines were also overexpressed in the lungs of F(1) littermates compared with the control F(1) litters at pnd(21). Lineage-specific hematopoietic growth factors may play an important role in the transmission of neutrophil-associated disease susceptibility across generations.

Role of TNF-α in prenatal alterations in dams of mice under thermal stress

The possible involvement of cytokines such as tumour necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interferon-γ (IFN-γ) that are suspected of causing pregnancy loss and miscarriage has been investigated in dams of mice subjected to hyperthermia. Thermal stress was induced by exposing mice dams at 40±2°C for 4 h every day during the different phases of the gestation period whereas the normothermic animals were housed at 22±2°C. The effect of maternal thermal stress was measured in pregnant mice at different phases of the gestation period namely, blastogenesis-implantation phase (days 0–5 postconceptionem [p.c.]), organogenesis or embryogenesis phase (days 6–15 p.c.) and fetogenesis phase (days 16–20 p.c.). Uterine examination of dams subjected to hyperthermia on days 6–15 p.c. showed maximum reduction in live fetus number, gestational index and maximum preand postimplantation loss in comparison with dams housed in normothermic environment and dams exposed to thermal stress between days 0–5 and 16–20 p.c. Maximum resorption rate and number of non-viable fetuses were observed in dams exposed to hyperthermia during days 6–15 p.c. Elevated levels of TNF-α and IL-1β were observed in the amniotic fluid of dams subjected to hyperthermia during days 6–15 p.c. but IFN-γ levels remained unaltered. Single intraperitoneal (i.p.) administration of recombinant mouse TNF-α at a dose of 1 and 0.5 ng/mice in dams on day 6 in normothermic condition resulted in a reduced number of live fetuses. Administration of anti-TNF-α antibody i.p. at a dose of 10 μg/dam on day 6 p.c. and subjected to thermal stress between days 6–15 p.c. increased marginally the number of fetuses but failed to attain statistical significance in comparison with days 6–15 p.c. thermally stressed dams without antibody treatment. It is concluded that the induction of TNF-α, in the amniotic fluid is associated with thermal stress during pregnancy and may be linked to the reproductive performances of dams. This study will help in understanding the mechanism of thermal injury in pregnant subjects.