Category Archives: Cissus

Cissus Inhibits Bone Loss in Mice

Abstract

Inhibition of Bone Loss by Cissus quadrangularis in Mice: A Preliminary Report.

Women drastically loose bone during and after menopause leading to osteoporosis, a disease characterized by low bone mass increasing the risk of fractures with minor trauma. Existing therapies mainly reduce bone resorption, however, all existing drugs have severe side effects. Recently, the focus is to identify alternative medicines that can prevent and treat osteoporosis with minimal or no side effects. We used Cissus quadrangularis (CQ), a medicinal herb, to determine its effects on bone loss after ovariectomy in C57BL/6 mice. Two-month old mice were either sham operated or ovariectomized and fed CQ diet. After eleven weeks, mice were sacrificed and the long bones scanned using pQCT and μCT. In the distal femoral metaphysis, femoral diaphysis, and proximal tibia, control mice had decreased cancellous and cortical bone, while CQ-fed mice showed no significant differences in the trabecular number, thickness, and connectivity density, between Sham and OVX mice, except for cortical bone mineral content in the proximal tibia. There were no changes in the bone at the tibio-fibular junction between groups. We conclude that CQ effectively inhibited bone loss in the cancellous and cortical bones of femur and proximal tibia in these mice.

Banu J, Varela E, Bahadur AN, Soomro R…
J Osteoporos 2012
PMID: 22779034 | Free Full Text

The full study is available using the link above.

CQ may primarily attenuate bone resorption in OVX mice through the downregulation of proinflammatory cytokines but it does not rule out the possibility that it may also act through other pathways. There are reports that CQ also enhances bone mineralization by accumulating mucopolysaccharides at the site of bone formation [The following popper user interface control may not be accessible. Tab to the next button to revert the control to an accessible version.Destroy user interface control14]. Moreover, CQ is reported to increase calcium uptake and mechanical properties of bone in rats [The following popper user interface control may not be accessible. Tab to the next button to revert the control to an accessible version.Destroy user interface control15]. Phytochemical analyses of CQ show the presence of high levels of calcium, vitamin C, β-carotene [The following popper user interface control may not be accessible. Tab to the next button to revert the control to an accessible version.Destroy user interface control38, The following popper user interface control may not be accessible. Tab to the next button to revert the control to an accessible version.Destroy user interface control39], and flavanoids [The following popper user interface control may not be accessible. Tab to the next button to revert the control to an accessible version.Destroy user interface control25] some of these substances have established beneficial properties on bone. In vitro studies have shown that ethanolic extracts of CQ increased mRNA and proteins related to the bone formation pathway and IGF-I, IGF-II, and IGF binding protein [The following popper user interface control may not be accessible. Tab to the next button to revert the control to an accessible version.Destroy user interface control40, The following popper user interface control may not be accessible. Tab to the next button to revert the control to an accessible version.Destroy user interface control41]. More investigations are necessary to elucidate the mechanism(s) by which CQ influences bone metabolism. However, it is very encouraging to note that in studies done with CQ using very high doses (5000mg/kgbodyweight) [The following popper user interface control may not be accessible. Tab to the next button to revert the control to an accessible version.Destroy user interface control9] have not reported any toxic side effects. In the present study, we have used only 500mg/kgbodyweight of CQ and observed that the liver, spleen, and kidney weights were not altered significantly, suggesting that CQ may not have any severe side-effects….

We conclude that CQ can reduce OVX induced bone loss and it does this in the long bones in a site-specific manner with more effects on the cancellous bone of femur followed by tibia. CQ probably reduces bone resorption primarily by downregulating proinflammatory cytokines that are often increased after ovariectomy. The beneficial effects of CQ are probably due to the flavanoids present.

 

Cissus as a Phytoestrogen-Rich Fraction Compared to Estrogen in Rats

Abstract

Antiosteoporotic activity of phytoestrogen-rich fraction separated from ethanol extract of aerial parts of Cissus quadrangularis in ovariectomized rats.

Cissus quadrangularis L. (C. quadrangularis L.) (Vitaceae) has been reported in Ayurveda for its antiosteoporotic activity. The study separated the phytoestrogen-rich fraction (IND-HE) from aerial parts of C. quadrangularis L. and evaluated its effect on osteoporosis caused by ovariectomy in rats.
IND-HE was separated from the ethanol extract of C. quadrangularis. Ovariectomized female Wistar rats were divided into four groups (n = 6). Group 1: Control (distilled water), Group II: IND-HE (75 mg/kg p.o.), Group III: IND-HE (100 mg/kg p.o.) were treated once daily for 8 weeks and Group IV: standard estradiol group, received estrogen (1 mg/kg, s.c. bi-weekly). The effects on body weight were determined. DEXA (Dual energy-emission X-ray absorptimatory analysis) of whole body bone and femur was carried out. Blood was removed and analyzed for biochemical parameters. After sacrificing the animals, biomechanical study of right tibia and histopathology of pelvic bone was carried out.
IND-HE showed presence of phytoestrogen-rich fraction. IND-HE (75 and 100 mg/ kg) and estrogen treatment showed statistically significant increase in bone thickness, bone density and bone hardness. IND-HE (75 and 100 mg/kg) and estrogen treatment significantly increased serum estradiol. IND-HE (100 mg/kg) (P<0.05) and estrogen treatment increased serum vitamin D3 and serum calcium compared to control. Alkaline phosphatase was significantly reduced by IND-HE (100 mg/kg p.o.) and estrogen treatment. Histopathology and DEXA results indicated that IND-HE (75 and 100 mg/kg) prevented bone loss.
These findings confirm that phytoestrogen-rich fraction (IND- HE) possess good antiosteoporotic activity.

Aswar UM, Mohan V, Bodhankar SL
Indian J Pharmacol May 2012
PMID: 22701244 | Free Full Text

Cissus May Increase Local IGF-1 in Osteoblasts

Abstract

Cissus quadrangularis augments IGF system components in human osteoblast like SaOS-2 cells.

Osteoporosis is a public health problem which is associated with significant morbidity and mortality. Growth factors are produced locally in the bone and control cellular events such as induction of bone growth. Signaling through the Insulin-like growth factor (IGF)-I receptor (IGF-IR) by locally synthesized IGF-I or IGF-II in osteoblast is considered crucial for normal development and for bone remodeling. Traditional use of Cissus quadrangularis (C. quadrangularis) in the treatment of bone disorders have been documented, however its regulatory effects on IGF system components remain largely unknown. The present study is employed to delineate the effects of ethanolic extract of C. quadrangularis on the regulation of IGF system components in human osteoblast like SaOS-2 cells. RT-PCR analysis revealed an increase in the mRNA expression of IGF-I, IGF-II, IGF-IR in cells treated with C. quadrangularis when compared with control cells. The mRNA expression of IGF binding protein-3 (IGFBP-3) did not differ significantly between control and C. quadrangularis treated cells. Immunoradiometric analysis revealed increased levels of IGF-I, IGF-II and IGFBP-3 in the conditioned medium of C. quadrangularis treated cultures when compared with control. Western blotting analysis revealed increase in protein levels of IGF-IR in cells treated with C. quadrangularis. These results indicate positive regulation of C. quadrangularis on the IGF system components of human osteoblast like SaOS-2 cells.

Muthusami S, Ramachandran I, Krishnamoorthy S, Govindan R…
Growth Horm. IGF Res. Dec 2011
PMID: 22015109

Cissus Comparable to Raloxifine in Rats

Abstract

Effect of Cissus quadrangularis Linn on the development of osteopenia induced by ovariectomy in rats.

The aim of our study was to see the efficacy of petroleum ether extract of Cissus quadrangularis (CQ) on development of osteopenia in ovariectomy induced Wistar rats.
The female Wistar rats were ovariectomized or Sham operated. The rats were anesthetized with pentobarbital sodium (40 mg/ kg b.w, i.p.), the ovaries were removed bilaterally. Sham-operation was performed in the same manner but only exposing the ovaries (sham operated (SHAM) group). A day later, the ovariectomized rats were randomly divided into four groups of eight animals each. The groups are 1. Sham operated (SHAM), 2. Ovariectomized (OVX), 3. Ovariectomized and treated with 25 mg/kg b.w of raloxifene (OVX+RAL), 4. Ovariectomized and treated with 500 mg/kg b.w of petroleum ether extract of CQ (OVX+CQ). The treatment continued for 30 days. At the end of the treatment, rats in all groups were sacrificed by cervical dislocation. Before sacrifice, blood was collected for the estimation of serum ALP, TRAP, Calcium and hydroxyproline; where as the left femur was used for histomorphometrical analysis.
The findings assessed on the basis of animal weight, morphology of femur, histomorphometry and biochemical analysis. As compared to SHAM group, OVX group animals showed a significant rise in serum ALP, TRAP and hydroxyproline levels at the end of 1 month following ovariectomy while no significant change was seen in the serum calcium levels. ALP and TRAP levels of OVX + RAL and OVX + CQ groups showed a further increase following administration of raloxifene and Cissus quadrangularis. The serum hydroxyproline content was found to be increased in the OVX + CQ compared to SHAM group. CQ significantly increased the thickness of both cortical (p <0.001) and trabecular bone (p <0.001).This action of CQ is comparable to action of Raloxifene. These data suggest a strong anti-osteoporotic activity of CQ.
The results confirm, at least in part, for the use of Cissus quadrangularis in folk medicine to treat osteoporosis.

Potu BK, Nampurath GK, Rao MS, Bhat KM
Clin Ter 2011
PMID: 21912817

Cissus Anabolic Actions on Bone Cells

Abstract

Effects of Cissus quadrangularis on the proliferation, differentiation and matrix mineralization of human osteoblast like SaOS-2 cells.

Osteoporosis is a public health problem which is associated with significant morbidity and mortality. The repair of bone defect is still a big challenge for orthopedic surgeons. Traditional use of Cissus quadrangularis (C. quadrangularis) in the treatment of bone disorders has been documented. The present study was employed to delineate the effects of ethanolic extract of C. quadrangularis on the proliferation, differentiation and matrix mineralization of human osteoblast like SaOS-2 cells. Lactate dehydrogenase assayed in the conditioned medium of control and C. quadrangularis treated cells did not differ significantly indicating that ethanolic extract of C. quadrangularis is nontoxic to osteoblastic cells. [(3)H] Thymidine incorporation assay revealed that C. quadrangularis treatment has increased the DNA synthesis of human osteoblastic SaOS-2 cells indicating increased proliferation of these cells. The data on alizarin red and ALP staining revealed increased matrix mineralization of human osteoblast like SaOS-2 cells. The study also revealed that the anabolic actions of ethanolic extract of C. quadrangularis in human osteoblast like cells are mediated through increased mRNA and protein expression of Runx2, a key transcription factor involved in the regulation of bone matrix proteins. Chromatin immunoprecipitation analysis revealed increased transcriptional activity of Runx2 on the promoter of osteocalcin after C. quadrangularis treatment. These results indicate positive regulation of C. quadrangularis on the proliferation, differentiation, and matrix mineralization of human osteoblast like SaOS-2 cells.

Muthusami S, Senthilkumar K, Vignesh C, Ilangovan R…
J. Cell. Biochem. Apr 2011
PMID: 21308732

Cissus, Vitex, and Allophylus May Increase Osteoblasts

Abstract

Anti-osteoporotic constituents from Indian medicinal plants.

The objective of this study was to determine the in vitro osteogenic activities of selected medicinal plants used traditionally in India. The compounds isolated from three plants viz. Allophylus serratus, Cissus quadrangularis and Vitex negundo were evaluated for their in vitro osteogenic activities. Primary cultures of osteoblasts were used to determine the effects of these components on osteoblast functions. Five of the fourteen compounds isolated led to increase in osteoblast differentiation and mineralization. These findings lend support to the use of Allophylus serratus, Cissus quadrangularis and Vitex negundo in traditional medicine.

Kumar M, Rawat P, Dixit P, Mishra D…
Phytomedicine Nov 2010
PMID: 20554183

Cissus Stimulates Osteoblastogenesis in Rats

Abstract

Petroleum ether extract of Cissus quadrangularis (Linn.) enhances bone marrow mesenchymal stem cell proliferation and facilitates osteoblastogenesis.

To evaluate the effects of the petroleum ether extract of Cissus quadrangularis on the proliferation rate of bone marrow mesenchymal stem cells, the differentiation of marrow mesenchymal stem cells into osteoblasts (osteoblastogenesis) and extracellular matrix calcification. This study also aimed to determine the additive effect of osteogenic media and Cissus quadrangularis on proliferation, differentiation and calcification.
MSCs were cultured in media with or without Cissus quadrangularis for 4 weeks and were then stained for alkaline phosphatase. Extracellular matrix calcification was confirmed by Von Kossa staining. marrow mesenchymal stem cells cultures in control media and osteogenic media supplemented with Cissus quadrangularis extract (100, 200, 300 microg/mL) were also subjected to a cell proliferation assay (MTT).
Treatment with 100, 200 or 300 microg/mL petroleum ether extract of Cissus quadrangularis enhanced the differentiation of marrow mesenchymal stem cells into ALP-positive osteoblasts and increased extracellular matrix calcification. Treatment with 300 microg/mL petroleum ether extract of Cissus quadrangularis also enhanced the proliferation rate of the marrow mesenchymal stem cells. Cells grown in osteogenic media containing Cissus quadrangularis exhibited higher proliferation, differentiation and calcification rates than did control cells.
The results suggest that Cissus quadrangularis stimulates osteoblastogenesis and can be used as preventive/ alternative natural medicine for bone diseases such as osteoporosis.

Potu BK, Bhat KM, Rao MS, Nampurath GK…
Clinics (Sao Paulo) 2009
PMID: 19841707 | Free Full Text

Cissus Reduces Bone Loss in Rats

Abstract

Evidence-based assessment of antiosteoporotic activity of petroleum-ether extract of Cissus quadrangularis Linn. on ovariectomy-induced osteoporosis.

The increasing incidence of postmenopausal osteoporosis and its related fractures have become global health issues in the recent days. Postmenopausal osteoporosis is the most frequent metabolic bone disease; it is characterized by a rapid loss of mineralized bone tissue. Hormone replacement therapy has proven efficacious in preventing bone loss but not desirable to many women due to its side-effects. Therefore we are in need to search the natural compounds for a treatment of postmenopausal symptoms in women with no toxic effects. In the present study, we have evaluated the effect of petroleum-ether extract of Cissus quadrangularis Linn. (CQ), a plant used in folk medicine, on an osteoporotic rat model developed by ovariectomy. In this experiment, healthy female Wistar rats were divided into four groups of six animals each. Group 1 was sham operated. All the remaining groups were ovariectomized. Group 2 was fed with an equivolume of saline and served as ovariectomized control (OVX). Groups 3 and 4 were orally treated with raloxifene (5.4 mg/kg) and petroleum-ether extract of CQ (500 mg/kg), respectively, for 3 months. The findings were assessed on the basis of animal weight, morphology of femur, and histochemical localization of alkaline phosphatase (ALP) (an osteoblastic marker) and tartrate-resistant acid phosphatase (TRAP) (an osteoclastic marker) in upper end of femur. The study revealed for the first time that the petroleum-ether extract of CQ reduced bone loss, as evidenced by the weight gain in femur, and also reduced the osteoclastic activity there by facilitating bone formation when compared to the OVX group. The osteoclastic activity was confirmed by TRAP staining, and the bone formation was assessed by ALP staining in the femur sections. The color intensity of TRAP and ALP enzymes from the images were evaluated by image analysis software developed locally. The effect of CQ was found to be effective on both enzymes, and it might be a potential candidate for prevention and treatment of postmenopausal osteoporosis. The biological activity of CQ on bone may be attributed to the phytogenic steroids present in it.

Potu BK, Rao MS, Nampurath GK, Chamallamudi MR…
Ups. J. Med. Sci. 2009
PMID: 19736603 | Free Full Text

Cissus Stimulates Fetal Bone in Rats

Abstract

Petroleum ether extract of Cissus quadrangularis (LINN) stimulates the growth of fetal bone during intra uterine developmental period: a morphometric analysis.

The aim of the present study was to analyze the effect Cissus quadrangularis plant petroleum ether extract on the development of long bones during the intra-uterine developmental stage in rats.
Pregnant rats (n=12) were randomly assigned into either a control group (n=6) or a Cissus quadrangularis treatment (n=6) group. Pregnant rats in the Cissus quadrangularis group were treated with Cissus quadrangularis petroleum ether extract at a dose of 500 mg/kg body weight from gestation day 9 until delivery. The animals in the control group received an equal volume of saline. Newborn pups were collected from both groups for alizarin red S – alcian blue staining to differentiate ossified and unossified cartilage. The ossified cartilage (bone) was morphometrically analyzed using Scion image software.
Morphometric analysis revealed that the percentage of the total length of ossified cartilage (bone) in pups born to treated dams was significantly higher (P<0.001- -0.0001) than that of the control group.
The results of the present study suggest that maternal administration of Cissus quadrangularis petroleum ether extract during pregnancy can stimulate the development of fetal bone growth during the intra-uterine developmental period.

Potu BK, Rao MS, Kutty NG, Bhat KM…
Clinics (Sao Paulo) Dec 2008
PMID: 19061006 | Free Full Text

Cissus May Increase Osteoblasts via MAPK

Abstract

Cissus quadrangularis extract enhances biomineralization through up-regulation of MAPK-dependent alkaline phosphatase activity in osteoblasts.

Cissus quadrangularis Linn. has been implicated as therapeutic agent for enhancing bone healing. Though its osteogenic activity has been suggested, the underlying mechanism still remains unclear. In the present study, the effects of ethanol extract of C. quadrangularis (CQ-E) on osteoblast differentiation and function were analyzed using murine osteoblastic cells. The results indicated that mRNA expressions of osteoblast-related genes were not affected by the CQ-E treatment. However, alkaline phosphatase (ALP) activity and the extent of mineralized nodules were significantly increased in treated cells compared with controls. The addition of an extracellular regulated kinase 1/2 inhibitor, a Jun N-terminal kinase 1/2/3 inhibitor and a p38 mitogen-activated protein kinase (MAPK) inhibitor resulted in significantly decreased ALP activity, preferentially by p38 MAPK inhibitor. These results suggested that CQ-E may regulate osteoblastic activity by enhancing ALP activity and mineralization process, and the increased ALP activity effect of CQ-E is likely mediated by MAPK-dependent pathway.

Parisuthiman D, Singhatanadgit W, Dechatiwongse T, Koontongkaew S
In Vitro Cell. Dev. Biol. Anim.
PMID: 19057968