To Explore the Effect of Xiangpi Shengji Ointment on the Apoptosis of Wound Cells in Model Rats after Anal Fistula Operation based on Fas/Fas L Pathway / 世界科学技术-中医药现代化

Objective To explore the effect of Xiangpi Shengji Ointment on wound healing and apoptosis-related Fas/Fas L pathway in model rats after anal fistula operation.Methods Thirty-six SD rats were used to construct the anal fistula model by using steel wire hanging line and indwelling for 30 days.After successful modeling,27 rats with anal fistula were randomly selected for"fistulectomy"to construct a postoperative wound model.After operation,the wound model rats were randomly divided into three groups,9 rats in each group,which were Xiangpi Shengji Ointment Group,Vaseline Group and Model Group,and the remaining 9 rats with anal fistula were sham operation group.The rats in the Xiangpi Shengji Ointment group were externally applied with Xiangpi Shengji Ointment gauze,while those in the Vaseline group were externally applied with Vaseline gauze.The rats in the model group were only disinfected and rinsed.No special treatment was given to the rats in the sham operation group.The wound healing was observed on the 3rd,5th,7th and 10th day after medication intervention,and the wound healing rate was calculated.After 10 days of continuous intervention,wound tissues were taken from each group,and the histopathological changes,the number of apoptosis,the expressions of Fas,Fas L,caspase-8 and cyto-c in wound tissues were observed by HE staining,TUNEL staining and immunohistochemistry respectively,and the mRNA expressions of Fas,Fas L and cyto-c in wound tissues were detected by RT-PCR.Results Compared with the model group,Xiangpi Shengji ointment group and Vaseline group significantly promoted wound healing at 7 and 10 days after intervention(P<0.01),and the wound healing rate of Xiangpi Shengji ointment group was significantly higher than that of Vaseline group(P<0.01).After 10 days of drug intervention,compared with sham operation group,the apoptosis rate of Xiangpi Shengji ointment group,Vaseline group and model group increased significantly(P<0.01),and the relative expressions of Fas,Fas L and cyto-c mRNA and the expression levels of Fas,Fas L,caspase-8 and cyto-c protein in wound tissue increased significantly(P<0.05,P<0.01).Compared with the model group,the apoptosis rate,the relative expression of Fas,Fas L and cyto-c mRNA and the expression level of Fas,Fas L,caspase-8 and cyto-c protein in Xiangpi Shengji ointment and Vaseline groups decreased significantly(P<0.05,P<0.01).Compared with Vaseline group,the apoptosis rate of Xiangpi Shengji ointment group decreased significantly(P<0.01),and the relative expression of Fas,Fas L and cyto-c mRNA and the expression level of Fas,Fas L,caspase-8 and cyto-c protein decreased significantly(P<0.05,P<0.01).Conclusion Xiangpi Shengji Ointment can inhibit the activation of Fas/Fas L pathway,reduce the apoptosis of wound tissue cells and promote wound healing after anal fistula operation.

Experimental study of treasure pill pretreatment on Fas and Fas L of neonatal rats hypoxic brain injury / 中国免疫学杂志

To investigate the effect of treasure pill pretreatment on hypoxic brain injury Fas and Fas L protein expression.Methods:Newborn 7d Wistar rats 200,male or female,were randomly divided into 5 groups(n=40):normal control group, model group 1h hypoxia(Ⅰm-12h,Ⅰm-24h,Ⅰm-48h,Ⅰm-72h),model group 4h hypoxia(Ⅳm-12h,Ⅳm-24h,Ⅳm-48h,Ⅳm-72h), hypoxic 1h treasure pill treatment group(ⅠT-14h,ⅠT-24h,ⅠT-48h,ⅠT-72h),hypoxic 4h treasure pill treatment group(ⅣT-14h,ⅣT-24 h ,ⅣT-48 h ,ⅣT-72 h ).The newborn Wistar rats were placed in hypoxia chamber of 8%O2+92%N2 ( V/V) for establishment the model of neonatal rats with hypoxia.Before modeling continuous 7 d once-daily oral administration of saline and treasure pill , during hypoxia also dose ,after hypoxia 12 h,24 h,48 h,72 h animals were sacrificed blood and brain tissues tested.By ELISA to detect the protein levels of Fas and Fas L in serum and brain tissue;by immunohistochemistry method to detect the expression of Fas and Fas L in rat brain tissue.Results:Compared with normal control group ,other four groups the protein content and protein expression of Fas and Fas L are in the 12h time points after hypoxic significantly increased ,and 24 h time point that pealed at statistically significant ( P<0.01 ) ,model group was significantly higher than the treatment group at each time point ;model groups in 48 h,72 h time point the protein expression and protein content of Fas and Fas L was less than 12 h time point but still significantly higher than the control group ( P<0.01 );treasure pill groups in 48 h,72 h time point the protein expression and protein content of Fas and Fas L was less than 12 h time point ,and slightly higher than the normal control group ( P<0.05 ).Conclusion: Treasures pill can cut hypoxic brain damage in rats,the positive expression of Fas and Fas L protein content and protein ,and thus the hypoxic brain injury has a protective effect.

Effects of simvastatin on renal expression of Fas/Fas-L in aged diabetic rats after uninephrectomy / 第二军医大学学报

Objective: To explore the effects of hydroxy-methyl-glutaryl coenzymeA reductase inhibitor (HMG-CoA RI) simvastatin on renal expression of apoptosis related gene Fas/Fas-L in aged diabetic rats after uninephrectomy. Methods: Elderly uninephrectomized SD rats were randomly divided into uninephrectomized control group (group C), streptozocin(STZ)-induced diabetic model group(group D), and diabetes + simvastatin group (group S). The apoptosis indices in renal tisses of three groups were detected by terminal deoxynucleotide mediated nick end labeling 12 weeks later. Immunofluorescence and real-time quantitative PCR were employed to observe the expression changes of Fas/Fas-L genes at the transcription and translation levels. Results: Compared with group C, group D had significantly higher apoptosis index and expression level of Fas/Fas-L; the above parameters were lower in group S compared with those in group D (P<0.05). The expression of Fas/Fas-L protein was mainly in the renal tubules. Conclusion: Both Fas and Fas-L genes are associated with the development and progression of diabetic nephropathy in aged rats. Simvastatin can down-regulate the translation and transcription of the genes, inhibit cell apoptosis and protect the kidney.

Absence of Fas-L aggravates renal injury in acute Trypanosoma cruzi infection

Trypanosoma cruzi infection induces diverse alterations in immunocompetent cells and organs, myocarditis and congestive heart failure. However, the physiological network of disturbances imposed by the infection has not been addressed thoroughly. Regarding myocarditis induced by the infection, we observed in our previous work that Fas-L-/- mice (gld/gld) have very mild inflammatory infiltration when compared to BALB/c mice. However, all mice from both lineages die in the early acute phase. Therefore, in this work we studied the physiological connection relating arterial pressure, renal function/damage and cardiac insufficiency as causes of death. Our results show that a broader set of dysfunctions that could be classified as a cardio/anaemic/renal syndrome is more likely responsible for cardiac failure and death in both lineages. However, gld/gld mice had very early glomerular deposition of IgM and a more intense renal inflammatory response with reduced renal filtration, which is probably responsible for the premature death in the absence of significant myocarditis in gld/gld.

Involvement of Bcl-2 Family and Caspases Cascade in Sodium Fluoride-Induced Apoptosis of Human Gingival Fibroblasts

Sodium fluoride (NaF) has been shown to be cytotoxic and elicit inflammatory response in human. However, the cellular mechanisms underlying NaF-induced cytotoxicity in periodontal tissues have not yet been elucidated. This study is aimed to investigate the mechanisms of NaF-induced apoptosis in human gingival fibroblast (HGF). NaF decreased the cell viability of HGF in a dose- and time-dependent manner. NaF gave rise to apoptotic morphological changes including cell shrinkage, chromatin condensation, and DNA fragmentation. However, NaF did not affect the production of ROS. In addition, NaF augumented cytochrome c release from mitochondria into the cytosol, and enhanced caspase -9 and -3 activities., cleavage (85 kDa fragments) of poly (ADP-ribose) polymerase (PARP) and upregulation of voltage-dependent anion channel (VDAC) 1. These results demonstrated that NaF-induced apoptosis in HGF may be mediated with mitochondria. Furthermore, NaF elevated caspase-8 activity and upregulated Fas-ligand (Fas-L), suggesting involvement of death receptor mediated pathway in NaF-induced apoptosis. Expression of Bcl-2, an anti-apoptotic Bcl-2 family, was downregulated, whereas expression of Bax, a pro-apoptotic Bcl-2 family, was not affected in NaF-treated HGF. These results suggest that NaF induces apoptosis in HGF through both mitochondria- and death receptor-mediated pathway mediated by Bcl-2 family.

Effects of Erbie Decoction on the hepatocyte apoptosis-related factors Fas/Fas-L in rat with liver fibrosis / 中华中医药杂志

Objective: To investigate effects of Erbie Decoction on the hepatocyte apoptosis-related factors Fas/Fas-L in rat with liver fi brosis,and to explore its anti-fi brosis mechanism. Methods: To Induce immune Liver fibrosis of rat model were established by intraperitoneal injection of pig serum. 48 rats were randomly divided into normal group, model group, pretreatment groups(high, middle and low dose groups of Erbie Decoction, and Fufang Biejia Ruangan Pills group. One week after the feeding adaptation, in addition to the normal group, other groups were given intraperitoneal injection of pig serum, each rat with 0.5ml, twice a week. At the same time, pretreatment groups were given intragastric administration daily. After 9 weeks, the expression of TGF-?1 and Fas/Fas-L were detected with immunohistochemistry assay. Results: Compared with normal group, in model group, TGF-?1 and Fas/Fas-L increased obviously(P

The Clinical Significance of Fas and Fas Ligand Expression in Thyroid Neoplasms

PURPOSE: Recent studies indicate that Fas and Fas Ligand (Fas-L) are implicated in autoimmune endocrine diseases and tumors of the thyroid. In this study we tried to elucidate the expression stati of Fas and Fas-L in some kinds of thyroid neoplasms, and their relationships with 4 prognostic factors in papillary thyroid cancer (i.e., size, lymph node metastasis, capsule invasion, age). METHODS: 66 cases of thyroid neoplasm including 45 cases of papillary cancer (PTC), 3 of a follicular cancer (FTC), 1 of a poorly differentiated cancer (PDC), 1 of a undifferentiated cancer (UC), 7 of follicular adenoma (FA), and 9 of nodular hyperplasia (NH) were examined, and estimated as negative, weak positive and strong positive about the Fas and Fas-L expression by the immunohistochemical staining intensities. We then collected and compared the differrences between benign and malignant tumors. The expressions of Fas and Fas-L in papillary thyroid cancers were evaluated relating to the differences in the prognostic factors (i.e., the size, lymph node status, capsule invasion, and age of the patients). RESULTS: Malignant thyroid tumors revealed stronger staining intensity than benign neoplasms. In papillary thyroid cancers, Fas-L staining intensities were significantly stronger in the cases with perithyroidal lymph node metastasis, or in those of 45 years old or over than in those with no lymph node metastasis, or younger than 45 years. CONCLUSION: Both Fas and Fas-L are implicated in thyroid tumorigenesis and revealed stronger staining intensities in malignant than benign tumors, and the Fas-L staining intensities may have some prognostic implications at least in papillary thyroid cancers.

A Study of Fas / Fas - Expression and Apoptosis according to the Progression of Gastric Adenocarclnoma / 대한암학회지

PURPOSE: The purpose of this study was to determine whether Fas-L expression is associated with increased apoptotic induction of tumor-infiltrating lymphocytes (TIL) in human gastric carcinomas. MATERIALS AND METHODS: The author analysed 38 cases of early gastric carcinoma (EGC) and 61 cases of advanced gastric carcinoma (AGC) who received gastric resection, in whom the number of diffuse type was 38 cases and the number of intestinal type was 61 cases. The author used immunohistochemical staining for Fas, Fas-L and CD45, and TUNEL in situ apoptosis detection kit. TIL were detected by CD45 and apoptosis of TIL were detected by CD45 expression and TUNEL positivity on serial histologic sections. RESULTS: Fas-L was localized to neoplastic cells in 61% (23/38) of EGC group and 66% (40/61) of AGC group. The extent of Fas-L expression was variable, with both Fas-L positive and negative neoplastic region occuring within tumors. TIL adjacent to Fas-L expressing tumor region were decreased in number and TIL adjacent to FasL-negative tumor region were increased in number; apoptotic induction of TIL showed just the opposite pattern (p<0.05). Fas expression was found essentially homogeneously throughout the tumor mass independent of tumor stage. Fas expression showed 64% (39/61) of intestinal type and 68% (26/38) of diffuse type. Labeling indices for tumoral apoptosis in EGC and AGC were 6.72% and 7.13%, respectively and this difference was statistically insignificant. Co-expression of Fas-L and Fas, which occurred over large areas of the tumors, did not result in an enhanced rate of tumor cell apoptosis. In addition, factors such as tumor stage and other prognostic factors were not concerned in Fas and Fas-L expression, number of TIL and apoptotic induction. CONCLUSION: These findings suggest Fas-mediated apoptotic depletion of TIL in response to Fas-L expression by stomach cancers, and provide the evidence to support the Fas counterattack as a mechanism of immune escape in gastric cancer. In addition, gastric carcinoma cells of the intestinal and diffuse type did not differ in their expression of the apoptotic receptor Fas.

Effects of losartan on apoptosis and expression of Fas and Fas-L in the kidney of diabetic rats / 中华肾脏病杂志

Objective To investigate the effects of angiotensin II type I receptor antagonist losartan on apoptosis and the expression of Fas and Fas-L in the kidney of diabetic rats. Methods Uninephrectomized, STZ(65 mg/kg) -induced diabetic male SD rats were used. Losartan(40 mg/kg) was delivered daily by gavage from the next day of the induction to diabetes for 12 weeks. Apoptosis was examined by means of terminal-deoxynucleotidyl transferase mediated d-UTP nick end labeling(TUNEL) . Flow cytometry and immunohistochemistry were used to detect the expression of Fas and Fas-L. Results Compared with those in the kidneys of control group, apoptotic cells were more in number and the expression of Fas and Fas-L was higher in the diabetic kidneys( P

Effects of pumpkin polysaccharide on Fas、Fas-L、Bcl-2 and Bax expression in the pancreas of diabetic rats / 中国药理学通报

Aim To investigate the effects of pumpkin polysaccharide(PP) on Fas、Fas-L、Bcl-2 and Bax expression in the pancreas of diabetic rats.Methods Rats were injected intraperitoneally(ip) with streptozocin(STZ) 60 mg?kg-1 to induce diabetic model.The diabetic rats were given PP 150 mg?kg-1,300 mg?kg-1,600 mg?kg-1.The blood sample were taken after three weeks.Blood glucose was measured by glucose oxidase method;The expression of the Fas and Fas-L have been determined by using immunohistochemistry(S-P method);The expression of Bax mRNA and Bcl-2 mRNA were checked up by Reverse transcription PCR(RT-PCR).Results PP could decrease the level of blood glucose in streptozotocin-induced diabetic rats;After three weeks,The expressions of the Fas and FasL in ? cell of streptozotocin-induced diabetic rats were obviously lower;The expression of Bax mRNA was obviously down-regulated,the expression of Bcl-2 mRNA was obviously up-regulated,and the ratio of Bcl-2/Bax was advanced after the diabetic rats were given PP.Conclusions PP could decrease the level of blood glucose of the diabetic rats.The mechanism was related to decreasing the expressions of the Fas and FasL and regulating the expressions of Bcl-2 and Bax in the pancreas of diabetic rats.

The experimental study on the expression of Bcl-2 and Fas-L after trauma brain injury in rats / 中国法医学杂志

Objective To observe the alteration of Bcl-2 and Fas-L at various intervals after traumatic brain injury and study the relationship between the alteration and the post-injury interval. Method The rat brain contusion was incurred by falling impact injury, paraffin section was cut after the test group rats were killed after various survival interval and stained with antibody of Bcl-2 and Fas-L, the hemotoxylin and eosin (HE) staining was carried out meanwhile. The staining results were measured quantitatively with computer imaging analysis system. Results Positive staining nerve celis were observed around the contusion area. At 30min after injury, a few Bcl-2 could be found around the wound, the intensity and the quantity of Bcl-2 positive celis increased significantly as post-injury interval extended. At 4h, the intensity came to a peak. Then the staining decreased. Although some Fas-L positive staining celis could be found around the wound at 30min after injury, the staining increased insignificantly from Ih to 4h after the injury. After 4h, the Fas-L positive staining cell increased significantly both in intensity and in quantity as post-injury interval extended. Conclusion There is a rule that the expression of Bcl-2 and Fas-L alters along with post-injury interval extension, which will be of value in time estimation of brain injury.