Cyclophosphamide In Mice Research Articles

Optimization of extraction of flavonoids from Lonicera rupicola and analysis of its effects in resisting inflammation, relieving pain, enhancing immunity, and inhibiting pyroptosis

The present study optimized the extraction of flavonoids from Lonicera rupicola Hook. f. et Thoms(LRH) and explored its pharmacological effects, such as resisting inflammation, relieving pain, enhancing immunity, and inhibiting pyroptosis, aiming to provide data support and scientific basis for the development and utilization of LRH. Response surface methodology(RSM) was applied to optimize the extraction of flavonoids from LRH based on the results of single-factor experiments. Anti-inflammatory and analgesic effects of LRH flavonoids were evaluated via inflammation and pain models in mice, such as xylene-induced ear swelling, carrageenan-induced footpad swelling, writhing caused by acetic acid, and paw licking. The effect of LRH flavonoids on the carbon clearance index of monocytes and serum immunoglobulin A(IgA) and IgM levels was analyzed on the immunosuppression model induced by cyclophosphamide in mice. The anti-oxidative effect in vivo of LRH flavonoids on liver superoxide dismutase(SOD), catalase(CAT), and malondialdehyde(MDA) levels was determined based on the chronic/subacute aging model in mice induced by D-galactose. The levels of cysteinyl aspartate specific proteinase-1(caspase-1), interleukin-1β(IL-1β), and IL-18 in the supernatant of J774 A.1 mononuclear phagocytes were detected to evaluate the effect of LRH flavonoids on the pyroptosis of mononuclear phagocytes in mice induced by the combination of lipopolysaccharide(LPS) and adenosine triphosphate(ATP). Meanwhile, the effect of LRH flavonoids on the cAMP-PKA signaling pathway was also explored. The optimum conditions for the extraction of LRH flavonoids are listed below: extraction temperature of 65 ℃, the ethanol concentration of 50%, extraction time of 60 min, a material-liquid ratio at 1∶25, and the yield of LRH flavonoids of 0.553%. RSM determined the multiple quadratic regression equation model of response value and variables as follows: the yield of LRH flavonoids=0.61-0.48A+0.1B+0.029C-0.014D+0.32AB+0.04AC-0.012AD-0.02BC+0.037BD-0.031CD-0.058A~2-0.068B~2-0.069C~2-0.057D~2. LRH flavonoids could effectively inhibit ear swelling and footpad swelling, reduced acetic acid-induced writhing, and delayed the paw licking response time in mice. Additionally, LRH flavonoids could improve the carbon clearance index in immunosuppressed mice, potentiate the activities of SOD and CAT and reduce MDA levels in the liver of aging mice induced by D-galactose, and effectively inhibit macrophage pyroptosis by decreasing the levels of caspase-1, IL-1β, and IL-18. The results reveal that LRH flavonoids possess excellent pharmacological activities such as resisting inflammation and oxidation, relieving pain, and enhancing immunity. They can inhibit pyroptosis by enhancing the cAMP-PKA signaling pathway. The results of this study can underpin the pharmacological research, development, and utilization of LRH.

Celastrol induces caspase-dependent apoptosis of hepatocellular carcinoma cells by suppression of mammalian target of rapamycin.

To investigate the efficacy of celastrol treatment of hepatocellular carcinoma (HCC) cells in vitro and in vivo and to propose a mechanism of action. A human HepG2 liver cancer cell line and a xenograft tumor model were used to investigate the effects of celastrol on HCC in vitro and in vivo. A CCK-8 kit was used to detect cell viability. Flow cytometry and terminal-deoxynucleoitidyl transferase mediated nick end labeling staining were used to detect apoptosis. Western blotting and immunohistochemistry were used to detect the expression of cleaved-caspase-3, cleaved-caspase-8, cleaved-caspase-9, cleaved-PARP, mammalian target of rapamycin (mTOR), and p-mTOR. Hematoxylin-eosin staining was used to observe the tissue morphology. Celastrol decreased the viability of HepG2 cells and induced apoptosis. Western blot assays indicated that celastrol up-regulated cleaved-caspase-3, cleaved-caspase-8, cleaved-caspase-9, and cleaved-PARP by inhibiting the phosphorylation of mTOR in HepG2 cells. Moreover, celastrol inhibited the tumor growth in a xenograft model. Celastrol also induced caspase-dependent apoptosis (up-regulation of cleaved-caspase- 3, -8, -9, and cleaved-PARP) and inhibited the activation of mTOR in vivo. Celastrol induces caspase-dependent apoptosis in HCC cells by inhibiting the activation of mTOR.

The effect of glutathione and buserelin on the stereological parameters of the hypothalamus in the cyclophosphamide-treated mice

IntroductionNew anticancer drugs have increased the survival and fertility rates in young patients. These drugs (i.e., cyclophosphamide; Cyc) have some side effects on the hypothalamus and fertility. One possible chemical for reducing these side effects is thiol or GnRH agonist. This study aimed to evaluate the capability of these agents for reducing the cyclophosphamide effects on the hypothalamus. MethodsSixty-three female mice were randomly assigned into seven groups. All groups including the control group had free access to water and mouse chow ad libitum. The sham group received normal saline. The Glu and Bus groups received glutathione (Glu) and buserelin (Bus) daily for 16 days, while the Cyc group received only cyclophosphamide as a single dose; the Cyc + Glu and Cyc + Bus groups, in addition to cyclophosphamide, received glutathione and buserelin, respectively. The volume of the hypothalamus, its neuron number, and dead neurons were evaluated using stereological methods. ResultsThere was no significant difference in the evaluated stereological parameters between the control and sham groups. However, the animals which received Cyc showed a decrease in the volume of the hypothalamus and its neuron number and density and an increase in cell death as compared with the control group. The treatment of the mice that received Cyc with Glu or Bus prevented these changes. ConclusionThis study showed that both GnRH agonist and thiol preserved or improved structural changes in the hypothalamus caused by cyclophosphamide in mice, suggesting that using thiol and especially GnRH agonist along with chemotherapy drugs may have protective effects on fertility.

Ameliorative effect of low molecular weight peptides from the head of red shrimp (Solenocera crassicornis) against cyclophosphamide-induced hepatotoxicity in mice

• Low molecular-weight peptides were prepared from Solenocera crassicornis head. • SCHPs-F1 ameliorated hepatotoxicity induced by cyclophosphamide in mice. • SCHPs-F1 restored endogenous antioxidants levels by activating the Nrf2 signal. • SCHPs-F1 suppressed hepatic inflammation through NF-κB signal pathway. • SCPHs-F1 relieved CTX hepatotoxicity as functional component. Our study was aimed at investigating the protective effect of low molecular weight peptides (SCHPs-F1) from red shrimp ( Solenocera crassicornis) head against cyclophosphamide-induced hepatotoxicity in mice. We found that SCHPs-F1 treatment dose-dependently normalized the biochemical markers, hepatic index, and total CYP450 enzyme content in CTX-induced hepatotoxicity in mice. SCHPs-F1 also ameliorated the CTX-mediated structural disorders of the hepatic tissue. Western blot results suggested that SCHPs-F1 significantly restored the levels of endogenous antioxidants (CAT, T-AOC, GSH-Px, SOD and MDA levels) through activating the Nrf2 signal by upregulating the expression of GCLM, HO-1, and NQO-1. Moreover, SCHPs-F1 improved the CTX-induced hepatotoxicity by inhibiting of NF-κB signal responses and down-regulating the expression of the inflammatory factors (IL-1β, IL-6, IFN-γ and TNF-α). These findings suggest that SCHPs-F1 can regulate Nrf2 and NF-κB signaling pathways and reduce the oxidative stress and inflammation in CTX-induced hepatotoxicity. Overall, SCHPs-F1 are value-added food ingredients for alleviating CTX-induced hepatotoxicity.

Study on the Changes of Chemical Constituents in Different Compatibilities of Ginseng-Prepared Rehmannia Root and Their Effects on Bone Marrow Inhibition after Chemotherapy.

Ginseng (G) and Prepared Rehmannia Root (PRR) are commonly used in traditional Chinese medicine for blood supplementation. This study aimed to study G and PRR with different compatibility ratios changes in chemical composition and inhibition of cyclophosphamide-induced myelosuppression. HPLC was used to determine the chemical constituents of 13 ginsenosides, 5-hydroxymethylfurfural (5-HMF) and verbascoside in different proportions of G-PRR. Balb/c mice were injected intraperitoneally with cyclophosphamide (CTX) to induce bone marrow suppression. The effects of different proportions of G-PRR on peripheral blood, bone marrow nucleated cells, thymus and spleen index of myelosuppressed mice were analyzed. The results showed that the compatibility of G and PRR can promote the dissolution of ginsenosides, and the content of conventional ginsenosides decreased, and the content of rare ginsenosides increased. Different proportions of G-PRR increased the number of peripheral blood and bone marrow nucleated cells in cyclophosphamide-induced bone marrow suppression mice (p < 0.01), increased thymus index (p < 0.01), decreased spleen index (p < 0.01). Different proportions of G-PRR can improve the myelosuppression induced by cyclophosphamide in mice, and the combined effect of G-PRR is better than the single decoction of G and PRR. Among them, G-PRR 2 : 3 and G-PRR 1 : 2 were better than the other groups. These results indicate that different proportion of G-PRR can improve bone marrow suppression, and the combined decoction of G-PRR is better than the separate Decoction in improving bone marrow suppression. This improvement may be related to the changes of the substance basis and active ingredients of G-PRR.

Antitumour and anti-haematotoxic activity of Asparagus racemosus L total dissolved solids in co-administration with cyclophosphamide in mice

Cancer is the second largest killer in the world. Even though the treatment and diagnosis of cancers have increased manifold, the survival rate has not increased proportionately. This discrepancy is mainly due to the toxicity of the chemotherapeutic agents. Natural herbs possessing significant antitumour activity can be utilized to address this issue. Asparagus racemosus is a widely distributed medicinal plant having reported antitumor properties. A concomitant use of the aqueous extract of Asparagus racemous (AEAR) along with Cyclophosphamide (CTX) was studied for the reversal of myelosuppression and hemotoxicity induced by CTX and to test whether AEAR possess significant antitumor activity against Dalton’s Lymphoma Ascites (DLA) tumour in BALB/c mice model. The study consisted of preliminary physico-chemical character determination and phytochemical screening. AEAR was used at 216, 432 and 864 g/kg body weight doses designated as lower, medium and high doses respectively and CTX was used at 25 mg/kg body weight. Hemogram, total leucocyte count, differential leucocyte count, bone marrow cellularity and – esterase positive cells were assessed. Malondialdehyde (MDA), glutathione, superoxide dismutase (SOD) and catalase (CAT) levels were estimated as oxidative stress markers. Tumour volume, size, mean survival time, increase in life span and in vitro cytotoxicity assays involving trypan blue exclusion test and MTT assay were done for antitumor study. AEAR showed significant elevation in hemogram, total leucocyte count, differential count, bone marrow cellularity and -esterase positive cell activity. AEAR at medium and high dose levels showed significant elevation in anti-oxidant enzyme levels of MDA, Glutathione, SOD and CAT enzymes. They also reduced the tumour volume, size and significantly increased the life span and mean survival time of DLA bearing mice. In in vitro cytotoxicity tests also, the medium and high dose of AEAR showed significant cytotoxicity. AEAR at doses of 432 and 864 mg/kg body weight doses significantly reverses the myelosuppression and hemotoxicity. It enhances the levels of antioxidant enzymes and is also an antitumor agent and exerts significant cytotoxicity against DLA tumour cells. Asparagus racemosus provides a ray of hope in addressing the toxicity while using chemotherapeutic agents when used as an adjuvant therapy.

Protective effects of betulinic acid on intestinal mucosal injury induced by cyclophosphamide in mice.

Betulinic acid (BA) is a plant-derived pentacyclic triterpenoid with a variety of biological activities. The purpose of this study was to assess the potential protective role of BA against intestinal mucosal injury induced by cyclophosphamide (CYP) treatment. Mice were pretreated with BA daily (0.05, 0.5, and 5.0 mg/kg) for 14 days, then injected intraperitoneally with CYP (50 mg/kg) for 2 days. BA pretreatment reduced the contents of malondialdehyde (MDA) and glutathione (GSH), decreased the activity of superoxide dismutase (SOD) in small intestine, increased villus hight/crypt depth ratio and restored the morphology of intestinal villi in CYP-induced mice. Moreover, BA pretreatment could significantly down-regulate the levels of pro-inflammatory cytokines interleukin-5 (IL-5), IL-17, IL-12 (P70) and tumor necrosis factor α (TNF-α), reduced production of chemokines macrophage inflammatory protein-1α (MIP-1α), macrophage inflammatory protein-1β (MIP-1β) and regulated upon activation, normal T-cell expressed and secreted (RANTES), and enhanced the levels of anti-inflammatory such as IL-2 and IL-10 in serum, and decreased the mRNA expressions of IL-1β and TNF-α in intestine of CYP-induced mice. Furthermore, RT-PCR demonstrated that BA improved intestinal physical and immunological barrier in CYP-stimulated mice by enhancing the mRNA expressions of zonula occluden 1 (ZO-1) and Claudin-1. BA might be considered as an effective agent in the amelioration of the intestinal mucosal resulting from CYP treatment.

The effect of vitamin C supplementation on neutropenia induced by cyclophosphamide in mice

The effect of vitamin C supplementation on neutropenia induced by cyclophosphamide in mice

Hesperidin opposes the negative impact of cyclophosphamide on mice kidneys

The present investigation examined the prospective nephroprotective effect of hesperidin (HSN) in mice challenged with a single i.p. injection of cyclophosphamide (CPE) at a dose of 200 mg/kg. HSN (100 and 200 mg/kg/day, p.o.) was given for 10 days, starting 5 days prior to CPE administration. HSN significantly reduced the CPE-induced increments of serum creatinine and cystatin C. HSN also significantly reduced malondialdehyde, nitric oxide, Bax/Bcl-2 ratio, and caspase-3, and significantly raised total antioxidant capacity, and interleukin-10/tumor necrosis factor-α ratio in kidneys of mice received CPE. In addition, HSN significantly prevented the histopathological injury, and kidney injury molecule-1 expression in kidneys of mice given CPE. It was concluded that HSN guarded against nephrotoxic effect of CPE in mice by tackling oxidative/nitrative stress, inflammation, and apoptosis.

Effect of Capparis spinosa L. Leaf bud Extract on The Hematological and Histological Changes Induced by Cyclophosphamide in Mice

Medicinal plants have been suggested to be chemoprotective on account of antioxidants and antitoxic properties. Capparis spinosa L. one of the medicinal plants is widely used in folk medicine. Therefore, the present study aimed to find a protective efficacy of ethanolic and aqueous extract leaf bud of C. spinosa against toxic effects of cyclophosphamide (CP) in mice. Results showed significant decreasing (p=0.05) in red blood cells (RBCs) hemoglobin (HB), packed cell volume (PCV%) and platelets count (PLT) reach to 7.36×1012/L, 96.83g/L, 31.6% and 925.0×1012/L respectively, in blood of mice treated with CP compared with control group which was 8.58×1012/L, 122.33g/L, .44.41% and 2194.16×1012/L, respectively. Significant decreasing in the values of white blood cells number (WBCs), and percentage of lymphocyte and monocyte, reach to 3.36×109/L,26.61% and 4.86% in blood of mice treated with CP compared with control. Significant increasing were showed in some liver enzymes (AST, and ALT) when exposed to CP which reach to 166.0U/L and 63.16U/L respectively, compared with control group. While groups treated with extract showed non significant differences compared with control group in the values of blood parameters and enzymes. Histopathological changes in liver when mice treated with CP represent by dilated of central vein, swelling of hepatocyte, focal accumulation of inflammatory cells, excessive vacuolated of hepatocyte, glycogenic degeneration, congested of central vein with sinusoid, and vacuolated of cytoplasm of hepatocytes with irregular nucleolus. While kidney showed degeneration of epithelial cells as foaming lumen, Clear hemorrhagic interstitial some of renal tubules are closure with degeneration of epithelial cells, Congestion of blood vessels. The tissues of kidney showed cloudy swelling of epithelial cells and hemorrhages, some of cells were elongated towered lumen, and there were inflammatory cells in interstitial tissue. Results showed the important role of leaf bud extract of C. spinosa in keeping the normal values of blood parameters and prevent histological changes as control.

Tarin stimulates granulocyte growth in bone marrow cell cultures and minimizes immunosuppression by cyclo-phosphamide in mice.

Chemotherapeutic drugs, such as cyclophosphamide, cause severe immunosuppression and patients become susceptible to infections. Based on this, the immunomodulatory potential of tarin, a lectin from Colocasia esculenta, was evaluated in bone marrow cell cultures and in cyclophosphamide-immunosuppressed mice. Tarin promoted maintenance of hematopoietic progenitors and repopulation of Gr1 cells in vitro which was supported by in vivo results. In immunosuppressed mice, tarin increased bone marrow cell numbers and altered cell profile distribution by enhancing the frequency of Gr1+ progenitors, including Ly6-CintLy6-Glo, and anticipating their proliferation/differentiation in mature cells, especially Ly6-CloLy6-Ghi. Bone marrow cells harvested from tarin-treated immunosuppressed mice proliferated in response to GM-CSF or G-CSF in vitro and, the low numbers of bone marrow cells in the G0 phase, combined with a high number cells undergoing apoptosis confirmed that tarin promoted a faster and intense proliferation/differentiation, even in the presence of CY-induced toxicity. As a result, tarin minimized leukopenia in immunosuppressed mice promoting a faster recovery of peripheral leucocytes and protected erythroid bone marrow cells from CY-cytotoxicity in a dose-dependent manner. Data suggest that tarin could be considered a potential adjuvant to decrease leukopenia and possibly ameliorate anemia, if carefully evaluated in human cancer cell lineages and in clinical trials.

Ameliorative Effect of Dangguibuxue Decoction against Cyclophosphamide-Induced Heart Injury in Mice.

Dangguibuxue decoction (DBD), a kind of Chinese herbal medicine, has been widely used to treat blood deficiency disease in China. In this experiment, we studied the effects of the Dangguibuxue decoction (DBD) on the myocardial injury induced by cyclophosphamide in mice. Alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatine kinase (CK), and lactic dehydrogenase (LDH) in serum were detected by commercial kits. Total white blood cell (WBCs), platelets, and cytokines pathological changes of heart tissue were also examined. In addition, the protein levels of the NF-кB pathway were detected to reveal its mechanism. The results showed that DBD significantly decreased the levels of ALT, AST, CK, and LDH and increased WBCs in CTX-induced mice. In addition, DBD significantly alleviated pathological changes of heart tissue. DBD significantly reduced the protein expressions of NF-кB signaling pathway. In summary, DBD can be considered an effective drug to alleviate CTX-induced heart damage in mice.

Immunostimulating Effects of Enzyme Hydrolysate of Ginseng Marc Polysaccharides in Immune-suppressed Mice

Ginseng contains various health-beneficial bioactive compounds, such as ginsenosides and polysaccharides. Despite ginseng marc is produced after the extraction process and usually discarded as wastes, it still contains considerable amounts of potential bioactive compounds, including saponins and polysaccharides. Previously, we reported that glucan type ginseng oligosaccharides obtained by enzyme hydrolysis of ginseng marc-derived polysaccharides exhibit immunostimulatory activities in macrophages and, activated macrophages are in turn capable of inhibiting the growth of skin melanoma cells via induction of apoptosis. In the present study, an enzymatic hydrolysate (GEH) containing these ginseng oligosaccharides was prepared and immune-enhancing activities of GEH were evaluated in vivo using cyclophosphamide-treated immune-suppressed mice. Immunosuppression was induced by 3 day-intraperitoneal (i.p.) injection of cyclophosphamide in mice. When comparedh normal control group, the GEH administered orally for 29 days facilitated the recovery of weight gain, indices of spleen and thymus, and enhanced T lymphocyte and B lymphocyte proliferation, cytokine productions of IL-6, IL-4 and IL-10 in culture supernatants of Con A-treated splenic T lymphocytes, and increased the serum levels of IL-4 and IL-10 as well as NK cell activity. These results demonstrated that administration of GEH stimulates and enhances immune function in immune-suppressed mice. The results of this study suggest that GEH of ginseng marc can be developed as a health-beneficial food material with immunostimulatory activity.

Oligopeptide derived from solid-state fermented cottonseed meal significantly affect the immunomodulatory in BALB/c mice treated with cyclophosphamide

In this study, the immunomodulatory activity of oligopeptide (CP) derived from solid-state fermented cottonseed meal were investigated in immunosuppressed BALB/c mice models by treatment with cyclophosphamide (CY). Results indicated that oligopeptide increased the thymus and spleen indices of CY-treated mice. The count of plague forming cells (PFC) and the content of half serum hemolysis (HC50) in immunosuppressive mice were restored to the normal level in CP-10 and CP-20 groups while the cytokines interleukin (IL)-2, IL-6, and tumor necrosis factor alpha (TNF-α) were increased significantly in CP-20 group. Similar increasing the immunoglobulin of IgG and IgM content in the serum of CP-10 group were also observed. These findings indicated that oligopeptide derived from solid-state fermented cottonseed meal had a strong immune-enhancing activity as well as a protective effect against immunosuppression induced by cyclophosphamide in mice.

In vivo nonspecific immunomodulatory and antispasmodic effects of common purslane (Portulaca oleracea Linn.) leaf extracts in ICR mice

Ethnopharmacological relevancePortulaca oleracea (common purslane) is used in traditional medicine to cure various illnesses. However, its immune-protective properties and antispasmodic effects still need more pharmacological data if the plant will be utilized in herbal and drug formulations. Therefore, the present study determined the capacity of this plant species to modulate nonspecific immune responses and to confirm its antispasmodic activity in vivo in ICR mice. Materials and methodsPhagocytic activity of peritoneal macrophage, splenic lymphocyte proliferation and plasma lysozyme levels were measured in mice that were immunosuppressed using cyclophosphamide and treated with the ethyl acetate extract of Portulaca oleracea. In addition, the charcoal meal transit test was used to measure intestinal motility using ethanolic (EtOH), hexane (HEX), and ethyl acetate (EA) solvent extracts. Phytochemical analysis was undertaken and DPPH scavenging properties of the three solvent extracts were also determined. ResultsThe EA extract of P. oleracea exhibited immunoactivity through significant increase in phagocytosis and higher proliferative response in splenic lymphocytes. Plasma lysozyme level was also higher in EA-treated mice at high dose but this was not statistically significant. Decreased intestinal motility was also exhibited in mice treated with the three leaf solvent extracts compared to the negative control and the acetylcholine-treated group. The antispasmodic activity of the solvent extracts was comparable to that of the atropine-treated group. Phytochemical analysis showed the presence of tannins in EA extract in addition to alkaloids and steroids. The EtOH and HEX extracts contain alkaloids, steroids and terpenoids. DPPH scavenging activity was highest in the EA extract. ConclusionsThe present study showed that the EA extract of P. oleracea leaves ameliorated the immunosuppressive action of cyclophosphamide in mice. The results also indicated that the three solvent extracts of the plant decreased smooth muscle spasms in mice ileum. However, further experiments are warranted to further isolate the plant's immunoactive component. Also, the mechanisms involved in the immunoactivity and antispasmodic properties of P. oleracea deserve full elucidation.

Tribulus terrestris Protects against Male Reproductive Damage Induced by Cyclophosphamide in Mice.

Tribulus terrestris (TT) has been considered as a potential stimulator of testosterone production, which has been related with steroidal saponins prevailing in this plant. Cyclophosphamide (CP) is the most commonly used anticancer and immunosuppressant drug, which causes several toxic effects, especially on the reproductive system. Patients who need to use CP therapy exhibit reduced fertility or infertility, which impacts both physically and emotionally on the decision to use this drug, especially among young men. We hypothesized that the treatment with TT dry extract would protect the male reproductive system against CP toxicity. Mice received dry extract of TT (11 mg/kg) or vehicle by gavage for 14 days. Saline or CP was injected intraperitoneally at a single dose (100 mg/kg) on the 14th day. Animals were euthanized 24 h after CP administration, and testes and epididymis were removed for biochemical and histopathological analysis and sperm evaluation. The dry extract of TT was evaluated by HPLC analysis and demonstrated the presence of protodioscin (1.48%, w/w). CP exposure increased lipid peroxidation, reactive species, and protein carbonylation and altered antioxidant enzymes (SOD, CAT, GPx, GST, and GR). Moreover, acute exposure to CP caused a reduction on 17 β-HSD activity, which may be related to the reduction in serum testosterone levels, histopathological changes observed in the testes, and the quality of the semen. The present study highlighted the role of TT dry extract to ameliorate the alterations induced by CP administration in mice testes, probably due to the presence of protodioscin.

Flavonoid fraction of Cajanus cajan prohibited the mutagenic properties of cyclophosphamide in mice in vivo

Cajanus cajan (L.) is a Pigeon pea cultivated in tropical and subtropical areas. It contains many bioactive components. The present study aimed to assess the antimutagenic efficacy of a flavonoid fraction of Cajanus cajan (FFCC) to reduce cytotoxicity and genotoxicity induced by cyclophosphamide (CP). We assessed genotoxic and cytotoxic effects using chromosome aberration, in mouse bone-marrow cells and spermatocytes, cell viability and DNA damage, in mouse bone-marrow cells. Animals received FFCC at concentrations 50,100 and 200 mg/kg b wt by oral gavage, and injected simultaneously with CP (20 mg/kg b wt) for 24 h. The results revealed that FFCC was safe and its effect was normal compared to control group. Moreover, we observed significant inhibition of CP-induced chromosome abnormalities in both, somatic and germ, cells (p ≪ 0.05) after concurrent administration of different concentrations of FFCC and CP. FFCC reduced chromosome aberrations by 14.29%, 25.21% and 28.57% in somatic cells, and 25.35%, 35.21% and 49.29% in germ cells after simultaneous treatment with CP respectively. Additionally, FFCC improved the cell viability of bone-marrow cells in a concentration-dependent manner when administered concurrently with CP. Similarly, FFCC diminished DNA damage (p ≪ 0.05) in CP-treated animals. The inhibitory index of tail DNA (%) reached 90.6% at the highest concentration of FFCC when administered simultaneously with CP. In conclusion, the flavonoid extract improved cell viability and protected animal cells from the cytotoxic and genotoxic effects exhibited by CP. Cajanus cajan flavonoids might contain the antioxidant bioactivity that effectively lessened chromosome aberrations and DNA damage induced by mutagenic agents.

Dietary Chlorella vulgaris Ameliorates Altered Immunomodulatory Functions in Cyclophosphamide-Induced Immunosuppressive Mice.

Based on the well-known toxicity of cyclophosphamide (CYP) on the immune system, this research investigated the modulating effects of the long-term dietary Chlorella vulgaris (CV) supplementation on the immunosuppression induced by CYP in mice, in order to provide a novel dietary design to mitigate the side effects of CYP therapy. Control, CYP-treated, CYP + CV (6%), CYP + CV (12%) and CYP + CV (24%) were used for 6 weeks, CV supplement in diet recovered the significantly reduced immunological function in CYP treated mice. As CV may have a modulating function through the inducible expression of cytokines, we assayed the expressions of interleukin-2 (IL-2), interleukin-12 (IL-12), tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ). Our results suggested that CYP significantly reduced the lymphocytes proliferation and phagocytic activities of macrophages, and stimulated the production of IL-2, IL-12, TNF-α and IFN-γ and that this impairment has been successfully adjusted by CV supplementation. Treatment with the algae also enhanced the natural killer (NK) cells cytotoxicity, and ameliorate histological changes of the spleen in CYP-treated mice. Therefore, as we found in this study, a diet supplemented with whole CV has beneficial effects on CVP-induced immunosuppression, through its immunomodulatory potential.

Effects of Astragalus Combined with Angelica on Bone Marrow Hematopoiesis Suppression Induced by Cyclophosphamide in Mice.

Danggui Buxue Tang (DBT), a combination of Astragalus and Angelica at a 5 : 1 ratio, mainly promotes hematopoiesis. However, in the clinic, the combination ratio of Astragalus and Angelica to treat low hematopoietic function is not an absolute 5 : 1 ratio, suggesting that the herbs may promote hematopoiesis better after being combined at a certain range of ratios. The objective of this study is to investigate the effect of different ratio combinations of Astragalus and Angelica on bone marrow hematopoiesis suppression induced by cyclophosphamide (CTX) and to probe the interaction and mechanism of Astragalus combined with Angelica in promoting hematopoiesis. Following establishment of the model, mice were administered with Astragalus (6.00 g·kg-1), Angelica (3.00 g·kg-1), and combinations of Astragalus and Angelica at different ratios, including 10 : 1 (Astragalus 9.81 g·kg-1+Angelica 0.98 g·kg-1), 5 : 1 (Astragalus 9.00 g·kg-1+Angelica 1.80 g·kg-1), 2 : 1 (Astragalus 7.71 g·kg-1+Angelica 3.08 g·kg-1), 1 : 1 (Astragalus 5.40 g·kg-1+Angelica 5.40 g·kg-1), 1 : 2.5 (Astragalus 3.08 g·kg-1+Angelica 7.71 g·kg-1), 1 : 5 (Astragalus 1.80 g·kg-1+Angelica 9.00 g·kg-1), and 1 : 10 (Astragalus 0.98 g·kg-1+Angelica 9.81 g·kg-1). Our results suggested that Astragalus mixed with Angelica synergistically promoted hematopoiesis best when the combination ratio of Astragalus and Angelica was 1 : 1, 1 : 2.5 or 1 : 5; moreover, the effect of Angelica was greater than that of Astragalus. The potential mechanisms of the combinations of Astragalus and Angelica that promote hematopoiesis include the dissolution of the effective components, promoting the synthesis and secretion of hematopoietic growth factor (HGF) and the proliferation of hematopoietic progenitor cells (HPCs).

Protective Effects of Essential Oils as Natural Antioxidants against Hepatotoxicity Induced by Cyclophosphamide in Mice.

Clinical application of cyclophosphamide (CP) as an anticancer drug is often limited due to its toxicity. CP is metabolized mainly in the liver by cytochrome P450 system into acrolein which is the proximate toxic metabolite. Many different natural antioxidants were found to alleviate the toxic effects of various toxic agents via different mechanisms. Therefore, the present study aimed at investigating the role of essential oils extracted from fennel, cumin and clove as natural antioxidants in the alleviation of hepatotoxicity induced by CP through assessment of hepatotoxicity biomarkers (AST, ALT, ALP), histopathology of liver tissues as well as other biochemical parameters involved in the metabolism of CP. The data of the present study showed that treatment of male mice with cyclophosphamide (2.5 mg/Kg BW) as repeated dose for 28 consecutive days was found to induce hepatotoxicity through the elevation in the activities of AST, ALT, and ALP. Combined administration of any of these oils with CP to mice partially normalized the altered hepatic biochemical markers caused by CP, whereas administration of fennel, clove or cumin essential oils alone couldn’t change liver function indices. Moreover, CP caused histological changes in livers of mice including swelling and dilation in sinusoidal space, inflammation in portal tract and hepatocytes, as well as, hyperplasia in Kuppfer cells. However, co-administration of any of the essential oils with CP alleviated to some extent the changes caused by CP but not as the normal liver. CP was also found to induce free radical levels (measured as thiobarbituric acid reactive substances) and inhibited the activities of superoxide dismutase, glutathione reductase, and catalase as well as activities and protein expressions of both glutathione S-transferase (GSTπ) and glutathione peroxidase. Essential oils restored changes in activities of antioxidant enzymes (SOD, CAT, GR, GST, and GPx) caused by CP to their normal levels compared to control group. In addition, treatment of mice with CP was found to induce the protein expression of CYP 3A4, 2B1/2, 2C6, 2C23. Moreover, the present study showed that essential oils reduced the expression of CYPs 2E1, 3A4 but could not restore the expression of CYP 2C6 and 2C23 compared to CP-treated mice. Interestingly, pretreatment of mice with essential oil of clove was found to restore activities of DMN-dI, AHH, and ECOD which were induced by CP to their normal control levels. It is concluded that EOs showed a marked hepatoprotective effect against hepatotoxicity induced by CP. In addition, co-administration of CP with any of these oils might be used as a new strategy for cancer treatment to alleviate the hepatotoxicity induced by CP.