Animals ; Electrocoagulation ; Microwaves ; therapeutic use ; Pneumococcal Infections ; metabolism ; Proliferating Cell Nuclear Antigen ; metabolism ; Rabbits ; Spleen ; metabolism ; Splenectomy ; methods

OBJECTIVETo compare the effects of sanjie zhentong capsule (SZC) and danazol on rats with endometriosis (EMT).

METHODSTotally 48 adult female Lewis rats were selected, 12 as the blank control group, and the rest 36 rats in the estrus cycle were used to establish the EMT model. After modeling they were randomly divided into 3 groups, i.e., the model control group, the SZC treatment group, and the danazol treatment group, 12 in each group. Four weeks later the focus was measured by a second laparotomy. The normal saline at 1 mL/day was administered to rats in the model control group, SZC at 86.4 mg/day to those in the SZC treatment group, and danazol at 7.2 mg/day to those in the danazol treatment group. All the treatment lasted for 4 weeks. At the end of the treatment, a third laparotomy was performed to measure the size of focus. The expression of proliferating cell nuclear antigen (PCNA) was detected using immunohistochemical assay. The cell apoptosis rate was detected using terminal deoxynucleotidyl transferase-mediated deoxy-UTP nick-end labeling (TUNEL). The concentration of prostaglandin E2 (PGE2) in the peritoneal fluid and the serum were detected using ELISA.

RESULTSThere was statistical difference in the change of the focus volume between the SZC treatment group (-23.27 +/- 18.18) and the danazol treatment group (-12.28 +/- 10.04) and the model control group (13.97 +/- 7.54, P < 0.01). The expression of ectopic PCNA significantly decreased and the positive expression rate of TUNEL obviously increased in the two treatment groups when compared with the model control group (P < 0.05, P < 0.01). The expression of ectopic PCNA decreased and the positive expression rate of TUNEL increased more obviously in the SZC treatment group than in the danazol treatment group (P < 0.01). The concentration of PGE2 in the peritoneal fluid and the serum was significantly lower in the two treatment group when compared with the model control group (P < 0.01). The concentration of PGE2 in the peritoneal fluid and the serum was significantly lower in the SZC treatment group than in the danazol treatment group (P < 0.01).

CONCLUSIONSSZC and danazol both could inhibit the focus growth in EMT rats. SZC showed better effects. It was an effective drug for treating EMT.

Animals ; Capsules ; Danazol ; therapeutic use ; Dinoprostone ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Endometriosis ; drug therapy ; Female ; Proliferating Cell Nuclear Antigen ; metabolism ; Rats ; Rats, Inbred Lew ; Treatment Outcome

OBJECTIVETo assess the therapeutic effect of percutaneous intratumoral injection with lipiodol emulsion of chemotherapie agents (CALE) on implanted VX2 tumor in rabbits.

METHODSTwelve New Zealand rabbits with implanted VX2 tumor (24 models) were divided into lipiodol group, chemotherapeutic agent group and CALE group with intratumoral injections of the corresponding agents. The pathological changes of all the lesions were observed and the expression of proliferating cell nuclear antigen (PCNA) and vascular endothelial growth factor (VEGF) were evaluated 7 days after the operation.

RESULTSCompared with the lipiodol group and chemotherapie agent group, intratumoral injection of CALE resulted in the highest tumor necrosis rate and greatest tumor necrosis (P<0.01). The labeling indices of PCNA and VEGF expressions in CALE group were markedly lower than those in the other two groups (P<0.01).

CONCLUSIONPercutaneous intratumoral injection of CALE is an effective ablation approach for treatment of malignant solid tumors.

Animals ; Emulsions ; Injections, Intralesional ; Iodized Oil ; administration & dosage ; therapeutic use ; Neoplasms, Experimental ; metabolism ; pathology ; therapy ; Proliferating Cell Nuclear Antigen ; metabolism ; Rabbits ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo investigate the effects of intralesional steroid, interferon alpha-2b or verapamil injection on proliferation, apoptosis and TGF-beta1 expression in keloid and hypertrophic scar in vivo.

METHODS6 patients with keloids and 6 patients with hypertrophic scar were treated with intralesional injection of triamcinolone acetonide (40 mg/ml) or IFN alpha-2b (15 x 10(5) U/ml) or verapamil (2.5 mg/ml). Samples were collected on the 7th day after intralesional injection. Samples of untreated keloid and hypertrophic scar and normal skin were used as control. Expression of PCNA and TGF-beta1 was detected in situ by immunohistochemical staining, and apoptosis was detected in situ by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL).

RESULTS1) Triamcinolone acetonide could prohibit proliferative scars through inhibiting cell proliferation and TGF-beta1 expression, as well as inducing apoptosis. 2) IFN alpha-2b could prohibit proliferative scars through inhibiting cell proliferation and TGF-beta1 expression, but not inducing apoptosis; 3) Verapamil could also prohibit proliferative scars through inhibiting proliferation and TGF-beta1 expression in fibroblasts, as well as inducing apoptosis. While the effect of inducing apoptosis was stronger than that of triamcinolone acetonide, the effect of inhibiting TGF-beta1 expression was weaker than those of triamcinolone acetonide and IFN alpha-2b.

CONCLUSIONSAlthough intraleional injection of steroid, interferon alpha-2b or verapamil were all effective in the treatment of keloid and hypertrophic scar, their mechanisms are not similar.

Adolescent ; Adult ; Apoptosis ; Child ; Cicatrix, Hypertrophic ; drug therapy ; Female ; Humans ; Interferon-alpha ; therapeutic use ; Male ; Proliferating Cell Nuclear Antigen ; metabolism ; Recombinant Proteins ; Steroids ; therapeutic use ; Transforming Growth Factor beta1 ; metabolism ; Verapamil ; therapeutic use ; Young Adult

OBJECTIVE: To evaluate the protective function of Babao Dan (BBD) on 5-flurouracil (5-FU)-induced intestinal mucositis (IM) and uncover the underlying mechanism. METHODS: A total of 18 male mice were randomly divided into 3 groups by a random number table, including control, 5-FU and 5-FU combined BBD groups, 6 mice in each group. A single intraperitoneal injection of 5-FU (150 mg/kg) was performed in 5-FU and 5-FU combined BBD groups on day 0. Mice in 5-FU combined BBD group were gavaged with BBD (250 mg/kg) daily from day 1 to 6. Mice in the control group were gavaged with saline solution for 6 days. The body weight and diarrhea index of mice were recorded daily. On the 7th day, the blood from the heart of mice was collected to analyze the proportional changes of immunological cells, and the mice were subsequently euthanized by mild anesthesia with 2% pentobarbital sodium. Colorectal lengths and villus heights were measured. Intestinal-cellular apoptosis and proliferation were evaluated by Tunel assay and immunohistochemical staining of proliferating cell nuclear antigen, respectively. Immunohistochemistry and Western blot were performed to investigate the expressions of components in Wnt/β-catenin pathway (Wnt3, LRP5, β-catenin, c-Myc, LRG5 and CD44). RESULTS: BBD obviously alleviated 5-FU-induced body weight loss and diarrhea, and reversed the decrease in the number of white blood cells, including monocyte, granulocyte and lymphocyte, and platelet (P<0.01). The shortening of colon caused by 5-FU was also reversed by BBD (P<0.01). Moreover, BBD inhibited apoptosis and promoted proliferation in jejunum tissues so as to reduce the intestinal mucosal damage and improve the integrity of villus and crypts. Mechanically, the expression levels of Wnt/β -catenin mediators such as Wnt3, LRP5, β-catenin were upregulated by BBD, activating the transcription of c-Myc, LRG5 and CD44 (P<0.01). CONCLUSIONS BBD attenuates the adverse effects induced by 5-FU via Wnt/β-catenin pathway, suggesting it may act as a potential agent against chemotherapy-induced intestinal mucositis.
Animals ; Male ; Mice ; Antineoplastic Agents/therapeutic use* ; beta Catenin/metabolism* ; Diarrhea/drug therapy* ; Fluorouracil/pharmacology* ; Intestinal Mucosa ; Mucositis/metabolism* ; Pentobarbital/therapeutic use* ; Proliferating Cell Nuclear Antigen/metabolism* ; Saline Solution

OBJECTIVE: Physical exercise, a common non-drug intervention, is an important strategy in cancer treatment, including hepatocellular carcinoma (HCC). However, the mechanism remains largely unknown. Due to the importance of hypoxia and cancer stemness in the development of HCC, the present study investigated whether the anti-HCC effect of physical exercise is related to its suppression on hypoxia and cancer stemness. METHODS: A physical exercise intervention of swimming (30 min/d, 5 d/week, for 4 weeks) was administered to BALB/c nude mice bearing subcutaneous human HCC tumor. The anti-HCC effect of swimming was assessed in vivo by tumor weight monitoring, hematoxylin and eosin (HE) staining, and immunohistochemistry (IHC) detection of proliferating cell nuclear antigen (PCNA) and Ki67. The expression of stemness transcription factors, including Nanog homeobox (NANOG), octamer-binding transcription factor 4 (OCT-4), v-Myc avian myelocytomatosis viral oncogene homolog (C-MYC) and hypoxia-inducible factor-1α (HIF-1α), was detected using real-time reverse transcription polymerase chain reaction. A hypoxia probe was used to explore the intratumoral hypoxia status. Western blot was used to detect the expression of HIF-1α and proteins related to protein kinase B (Akt)/glycogen synthase kinase-3β (GSK-3β)/β-catenin signaling pathway. The IHC analysis of platelet endothelial cell adhesion molecule-1 (CD31), and the immunofluorescence co-location of CD31 and desmin were used to analyze tumor blood perfusion. SMMC-7721 cells were treated with nude mice serum. The inhibition effect on cancer stemness in vitro was detected using suspension sphere experiments and the expression of stemness transcription factors. The hypoxia status was inferred by measuring the protein and mRNA levels of HIF-1α. Further, the expression of proteins related to Akt/GSK-3β/β-catenin signaling pathway was detected. RESULTS: Swimming significantly reduced the body weight and tumor weight in nude mice bearing HCC tumor. HE staining and IHC results showed a lower necrotic area ratio as well as fewer PCNA or Ki67 positive cells in mice receiving the swimming intervention. Swimming potently alleviated the intratumoral hypoxia, attenuated the cancer stemness, and inhibited the Akt/GSK-3β/β-catenin signaling pathway. Additionally, the desmin⁺/CD31⁺ ratio, rather than the number of CD31⁺ vessels, was significantly increased in swimming-treated mice. In vitro experiments showed that treating cells with the serum from the swimming intervention mice significantly reduced the formation of SMMC-7721 cell suspension sphere, as well as the mRNA expression level of stemness transcription factors. Consistent with the in vivo results, HIF-1α and Akt/GSK-3β/β-catenin signaling pathway were also inhibited in cells treated with serum from swimming group. CONCLUSION Swimming alleviated hypoxia and attenuated cancer stemness in HCC, through suppression of the Akt/GSK-3β/β-catenin signaling pathway. The alleviation of intratumoral hypoxia was related to the increase in blood perfusion in the tumor. Please cite this article as: Xiao CL, Zhong ZP, Lü C, Guo BJ, Chen JJ, Zhao T, Yin ZF, Li B. Physical exercise suppresses hepatocellular carcinoma progression by alleviating hypoxia and attenuating cancer stemness through the Akt/GSK-3β/β-catenin pathway. J Integr Med. 2023; 21(2): 184-193.
Humans ; Animals ; Mice ; Carcinoma, Hepatocellular/drug therapy* ; Proto-Oncogene Proteins c-akt/metabolism* ; Proliferating Cell Nuclear Antigen/therapeutic use* ; Mice, Nude ; Glycogen Synthase Kinase 3 beta/genetics* ; beta Catenin/therapeutic use* ; Liver Neoplasms/drug therapy* ; Desmin/therapeutic use* ; Ki-67 Antigen ; Cell Line, Tumor ; Hypoxia ; RNA, Messenger/therapeutic use* ; Cell Proliferation

OBJECTIVETo study the effects of Xiongshao Capsule (XSC) on the proliferation of vascular smooth muscle cells (VSMC) in rabbits with experimental atherosclerosis (AS), and to explore its possible mechanisms.

METHODSRabbit's fractional AS model was established by denuding and injuring the endodermis of abdominal aorta with 4F * Fogarty catheter, followed with feeding of high cholesterol forage. The animals were randomly divided into 8 groups, the model groups of 3 days, 2 weeks and 6 weeks after modeling (Group A, B and C); the single endothelium injury group (Group D), the probucol treated group (Group E), the low-dose and high-dose XSC treated groups (Group F and G) and the sham operative group (Group N). All were fed with high fat forage except those in Group N and D. The proliferative activity of neogenetic SMC at abdominal aorta with the most obvious pathological changes was analyzed by proliferating cell nuclear antigen immunohistochemical method; the VSMC phenotypic modulation was detected by transmission electron microscope (TEM), and the level of plasma angiotensin (Ang II) was measured by radioimmunoassay. And the effects of treatment on them were observed as well.

RESULTSThe plasma Ang II level elevated gradually in Group A, and showed significant difference as compared that between Group C and Group N (P < 0.01); as compared with Group C, that in Group G was reduced significantly (P < 0.05); a reducing tendency was shown in Group E and F, but the difference of them with Group C was insignificant. Immunohistochemical dyeing showed that PCNA positive expressing cell was not found in the blood vessels of Group N, few was seen in Group A, while the upmost positive expression was shown in Group B, as for in Group C, significantly thickened endomembrane appeared, but the PCNA positive expression dropped. Number of PCNA positive cells reduced significantly (P < 0.05) in the drug treated groups, especially in Group G, showing significant difference as compared with that in Group C (P < 0.05, P < 0.01). TEM demonstrated normal shaped VSMC of aortic medial tunica in Group N, basically normal in Group A, but in Group B, migration of VSMCs into intima was found, as for in Group C, abundant lipid granules and bigger vacuoles appeared in VSMCs with markedly proliferated intercellular collagenous fibers. Synthetic transformation could also be found in Group D. The transform VSMCs in neogenetic endothelium was fewer in the drug treated groups than that in Group C. Morphology of VSMC was basically normal in Group G, with few intercellular collagenous fiber proliferation, while in Group E and F, many lipid droplets in VSMC and lot of collagenous fibers proliferation in intercellular space still retained.

CONCLUSIONSXSC can prevent the genesis and development of AS through significantly lowering the plasma Ang II level and inhibiting the migration and proliferation of VSMC.

Angiotensin II ; metabolism ; Animals ; Atherosclerosis ; drug therapy ; metabolism ; physiopathology ; Capsules ; therapeutic use ; Cell Proliferation ; drug effects ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Male ; Muscle, Smooth, Vascular ; cytology ; drug effects ; metabolism ; Proliferating Cell Nuclear Antigen ; metabolism ; Rabbits

OBJECTIVE: To study the anti- scarring effect of rapamycin in rabbits receiving glaucoma filtering surgery. METHODS: Ninety-six Chinchilla rabbits were randomized equally into 3 rapamycin treatment groups and one control group. All the rabbits underwent trabeculectomy, after which the rabbits in the 3 rapamycin groups were treated with eye drops containing 1%, 3%, or 5% rapamycin in the operated eyes, and those in the control groups were given castor oil 4 times a day. The intraocular pressure (IOP) and inflammatory reaction in the treated eyes were observed, and the PCNA-positive cells in the filtering bleb were detected using immunohistochemistry. RTFs isolated from the Tenon's capsule of the rabbits were cultured , and the expressions of caspase-3, caspase-8, and caspase-9 in the fibroblasts were detected after treatment with different concentrations of rapamycin. RESULTS: The IOP was significantly lower in rapamycin-treated group than in the control group after the surgery ( < 0.05). The counts of the PCNA-positive cells were significantly lower in rapamycin-treated rabbits than in the control group ( < 0.05). Rapamycin treatment dose-dependently increased the expressions of caspase-3 and caspase- 9 at both the mRNA ( < 0.001) and protein ( < 0.001) levels without causing significant changes in the expressions of caspase-8. CONCLUSIONS Rapamycin can inhibit excessive proliferation of the fibroblasts in the filtering bleb to reduce scar formation after glaucoma filtration surgery in rabbits. Rapamycin also increases the expressions of caspase-3 and caspase-9 to induce apoptosis of the RTFs.
Animals ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Proliferation ; drug effects ; Cicatrix ; prevention & control ; Filtering Surgery ; adverse effects ; Glaucoma ; surgery ; Intraocular Pressure ; Postoperative Complications ; enzymology ; prevention & control ; Proliferating Cell Nuclear Antigen ; analysis ; Rabbits ; Random Allocation ; Sirolimus ; therapeutic use ; Trabeculectomy

BACKGROUNDOuabain is a mammalian adrenocortical hormone that is involved in the pathogenesis of hypertension by inhibiting Na-K ATPase activity. It also participates in a variety of kinase-mediated signaling pathways associated with Na-K ATPase. Previous studies have shown that ouabain can cause cardiac remodeling independent of elevated blood pressure and that proliferating cell nuclear antigen (PCNA) plays a coordinating role for numerous proteins involved in multiple processes associated with DNA synthesis. Therefore, we hypothesized that ouabain might play a role in the cerebral cortex through signaling pathways independent of hypertension. And PCNA might be involved in this process.

METHODSMale Sprague-Dawley rats were treated with ouabain or with 0.9% nitric sodium as the control group. Systolic blood pressure was recorded weekly. After four weeks of treatment, morphological changes in the cerebral cortex were analyzed using light and transmission electron microscopy. The expression of PCNA in the cerebral cortex was evaluated by immunohistochemistry, real time quantitative PCR, and Western blotting.

RESULTSAfter 4-week treatment, there was no significant difference in systolic blood pressure compared with the control group, but both structural deterioration and up-regulated expression of PCNA in the brain was induced by ouabain treatment.

CONCLUSIONSThese results suggest that ouabain induces alterations in the brain structure, and this effect is independent of blood pressure. PCNA might be involved in the repair process of ouabain-induced brain damage.

Animals ; Blood Pressure ; drug effects ; Hypertension ; drug therapy ; metabolism ; Immunohistochemistry ; Male ; Ouabain ; therapeutic use ; Proliferating Cell Nuclear Antigen ; metabolism ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Sodium-Potassium-Exchanging ATPase ; metabolism

OBJECTIVETo dynamically observe the effect of Shenshuai Mixture (SM) on repair of kidney in acute renal failure (ARF) rats.

METHODSMale SD rats were divided into 4 groups randomly, the normal group, the SM group, the verapamil group and the model control group. Except those in the normal group were treated with normal saline without modeling, all remaining rats, after being made into ARF model by intra-muscular injection of glycerin, were treated with SM, verapamil and normal saline respectively via gastrogavage. Renal function, renal pathology, mRNA expression of epidermal growth factor (EGF) and protein expression of proliferating cell nuclear antigen (PCNA) were detected once every day from the 1st day to the 5th day. Results (1) BUN and Scr levels increased markedly 24 hrs after modeling, but the Scr level in the two treated groups was significantly lower than that in the model group (P < 0.05). Compared with that in the model group and the verapamil group, renal function was better in the SM group on the 3rd day (P < 0.01), and approach to normal level on the 5th day. (2) Renal pathological changes alleviated in every phase of ARF in the SM group than that in the model group, especially part of tubule regeneration could be seen on the 3rd day (metaphase), and renal structure was rehabilitated on the 5th day (convalescence), prior to those in the model group. (3) At the 3rd day expression of EGF mRNA and PCNA in tubule epithelial cell (TEC) increased remarkably in the SM group, higher than those in the model and verapamil group (P < 0.05).

CONCLUSIONSM could promote renal tissue regeneration and rehabilitation, and shorten the course of ARF through up-regulating mRNA expression of EGF in TEC.

Acute Kidney Injury ; drug therapy ; physiopathology ; Animals ; Drugs, Chinese Herbal ; therapeutic use ; Epidermal Growth Factor ; biosynthesis ; genetics ; Kidney ; physiopathology ; Male ; Phytotherapy ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley