Effects of therapeutic hypercapnia on inflammation and apoptosis after hepatic ischemia-reperfusion injury in rats

Background Therapeutic hypercapnia （TH） has been demonstrated to protect several organs ischemia-reperfusion injury.The study aimed to investigate the effects of therapeutic hypercapnia on hepatic ischemia-reperfusion injury （HIRI）.Methods Thirty adult male Wistar rats weighing （250 ± 20） g were randomized into 3 groups （n=10 in each）, group C （control group）, group A （hypercapnia group） and group B （CO2 preconditioning group）.A segmental ischemia of the liver was induced by interrupting the blood vessels including the bile duct to the median and left lateral lobes for 60 minutes and all the animals were sacrificed after 240 minutes observation period of reperfusion.Mean arterial pressure （MAP）and the blood gases were measured before ischemia （baseline） and at 30, 60, 120, 180 and 240 minutes after reperfusion.Arterial blood samples were obtained for determination of serum levels of TNF-α, IL-10, serum aspartate aminotransferase （AST） and alanine aminotransferase （ALT）.The histopathology of liver tissues was evaluated by light microscopy.The NF-κB expression and apoptotic hepatocytes were respectively determined by immunohistochemistry and TUNEL assay.Results The serum levels of liver enzymes and TNF-α were significantly decreased while the IL-10 level was significantly increased in groups A and B than in group C （P 〈0.05）, and group B surpassed group A （P 〈0.05）.The histopathological scores, the NF-κB immunohistochemical score （IHS） and apoptotic index were significantly lower in groups A and B than in group C （P 〈0.05）, and the decrease in group B was more obvious than in group A （P〈0.05）.Conclusion Therapeutic hypercapnia attenuates ischemia-reperfusion injury to the liver.Moreover, the effects of CO2preconditioning are outstandingly notable.     

Protective effect of diallyl trisulfide on liver in rats with sepsis and the mechanism

The protective effects of diallyl trisulfide on liver were examined in rats with sepsis. Sepsis was reproduced in rats by cecum ligation and puncture (CLP). Fifty-six male Wistar rats were randomly divided into sham-operated group (group S, n=8), sepsis model group (group C, n=24), diallyl trisulfide (DATS)-treated group (group D, n=24). Animals in groups C and D were further divided into three subgroups according to different observation time points, with 8 rats in each sub-group.Rats in group D and C were intravenously injected with normal saline or DATS respectively at a dose of 20 mg/kg after the establishment of sepsis model. Eight rats in groups C and D were sacrificed at 3, 6 and 24 h post-CLP and their livers were harvested for detection of interleukin (IL)-1 receptor associated kinase-4 (IRAK-4), nuclear factor-κB (NF-κB), c-fos, c-jun, malondialdehydethhe (MDA) and superoxide dismutase (SOD), tumor necrosis factor alpha (TNF-α) and for pathological examination. The results showed that the levels of serum IRAK-4, NF-κB and TNF-α in hepatic tissues were higher in group C than group S (control group) (P<0.05). After DATS treatment, the levels of IRAK-4 and NF-κB in the hepatic tissues and serum TNF-α in group D were lower than those in group C (P<0.05). The levels of c-fos and c-jun and MDA in the hepatic tissues were higher in group C than in group S (P<0.05). After DATS treatment, the levels of c-fos and c-jun and MDA in the hepatic tissues were significantly lower in group D than in group C (P<0.05). When compared with group S group, concentration of SOD in the hepatic tissues in group C was significantly lower (P<0.05). After DATS treatment, the concentration of SOD in the hepatic tissues was higher in group D than in group C (P<0.05). These findings suggested that treatment with DATS could ameliorate sepsis-induced liver injury in rats. The protective effect might be related to its ability to inhibit the signal pathway of IRAK-4 and NF-κB, thereby decreasing the production o     

Effect of ATRA on Contents of liver Retinoids, Oxidative Stress and Hepatic Injury in Rat Model of Extrahepatic Cholestasis

The effects of all-trans-retinoic acid （ATRA） administration on the concentration of retinoids （RA and vitamin A） in liver, oxidative stress and the hepatic injury in a rat model of common bile duct ligation （CBDL）-induced liver injury were investigated. Female rats were subjected to a sham （n=5） or CBDL （n=48）. Two weeks after operation, rats undergoing CBDL were randomized to receive treatment with either ATRA at three different doses （0.1, 1.5, 7.5 mg/kg） dissolved in bean oil or only bean oil every day over a 4-week experimental period. Rats were killed and blood samples were collected from the heart for determination of the serum transaminase. The contents of retinoids in rat liver were detected by using HPLC. Malondialdehyde （MDA）, glutathione （GSH） and superoxide dismutase （SOD） levels in liver were determined by a spectrophotometric method according to the instruction of the kits. Liver pathologic changes were observed under the light microscopy and electron microscopy. The results showed that compared with sham-operated group, the levels of retinoids in the liver tissue were significantly decreased in the CBDL group （P〈0.01）. ATRA （0.1 mg/kg） administration in CBDL rats partially restored the contents of retinoids （P〈0.05）. Liver RA and vita- min A contents in CBDL group were significantly increased after ATRA （1.5 and 7.5 mg/kg） supplementation as compared with sham-operated group （P〈0.05）. However, in ATRA-treated CBDL group, hepatic GSH level and SOD activity, depressed by CBDL, and hepatic MDA level, increased by CBDL were returned to those in sham-operated group （P〈0.05）. The histologic observation of liver tissues indicated that ATRA treatment notably alleviated hepatocellular swelling, steatosis, the. swelling of mitochondria and proliferation of smooth endoplasmic reticulum （SER）. Treatment with ATRA could reduce levels of serum transaminase as compared with sham-operated group, more greatly in 1.5 and 7.5 mg/kg ATRA-treated     

Low-dose ATRA Supplementation Abolishes PRM Formation in Rat Liver and Ameliorates Ethanol-induced Liver Injury

The effects of all-trans-retinoic acid (ATRA) in low doses supplementation on concentrations of polar retinoid metabolites (PRM) and retinoids in the ethanol-fed rat liver, and on hepato-cyte injury were investigated. The rat model of alcoholic liver disease (ALD) was induced by intra-gastric infusion of ethanol, and then the rats were administrated with ATRA in two different doses (150μg/kg body weight and 1.5 mg/kg body weight) for 4 weeks. Concentrations of retinoids in rat liver and plasma were determined by using HPLC. Liver tissues pathologic changes were observed under the light microscopy and electron microscopy. The serum transaminases concentrations were measured. The results showed that the HPLC analysis of retinoids revealed that retinoids (vitamin A, RA, retinyl palmitate) concentrations in ethanol-fed rat liver and RA concentration in ethanol-fed rat plasma were markedly diminished (P<0.01) after ethanol feeding for 12 weeks. Furthermore, obvious peaks of PRM were formed in livers of ethanol-fed rats. ATRA 150μg/kg supplementation in ethanol-fed rats for 4 weeks raised RA concentration in both liver and plasma, and also raised vitamin A concentration in liver to control levels, partially restored retinyl palmitate concentration (P<0.05) in liver. ATRA 1.5 mg/kg supplementation raised not only RA concentrations in liver and plasma but also retinyl palmitate concentrations in liver. However, the vitamin A concentration in liver of ATRA-supplemented rats (1.5 mg/kg) was higher than that of controls (P<0.05). The histologic observation of liver tissues indicated that ATRA treatment notably alleviated hepatocellular swelling, steatosis, the swelling of mitochondria and proliferation of smooth endoplasmic reticulum (SER). ATRA treatment greatly decreased levels of serum transaminases as compared with the only ethanol-fed group (P<0.05). It was concluded that low-dose ATRA treatment could restore retinoids concentrations and abolish the PRM formation in liver of ALD rats, and then ameliorate the injury of liver cells.     

Hepatoprotective activity of the ethanol extract of Sarcopyramis Nepalensis

The present study examined the protective effect of the ethanol extract of Sarcopyramis nepalensis (EESN) on agents-induced hepatotoxicity in mice and the possible mechanism. Acute liver injury was induced by administration of either CCl4 or D-GalN. The animals were divided into 5 groups in terms of different treatment: normal group, CCl4 or D-GalN group, silymarin or bifendate group, low dose EESN group (10 mg/kg) and high dose EESN group (30 mg/kg). Liver function was evaluated by detecting the levels of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST). The oxidize stress markers were measured, including malondialdehyde (MDA), glutathione peroxidase (GSH) and superoxide dismutase (SOD). Liver tissues were histopathologically examined by hematoxy-lin-eosin (H&E) staining. The acute toxicity study revealed that there was no toxicity of EESN at the dose of 5 g/kg in mice. The levels of ALT and AST in serum, and the MDA level in live tissues were significantly increased and the activities of SOD and GSH substantially decreased in mice after CCl4 or D-GalN treatment. These biochemical and oxidize stress markers were profoundly improved after treatment with EESN at different doses, which was similar to the results of silymarin or bifendate treatment. The histophathological examination revealed the significant improvement in the pathological changes of the liver in EESN-treated mice as compared to those in CCl4 or D-GalN group. It was concluded that EESN possesses potential antioxidant and hepatoprotective properties and has therapeutic potential for liver diseases.     

Effect of Kaiyu Qingwei Granule (开郁清胃颗粒) on Insulin Receptor in Liver and Skeletal Muscular Cell Membrane in Diabetes Mellitus Rats

Objective: To investigate the effect of Kaiyu Qingwei granule (KYQWG, on the insulin binding capacity of liver and skeletal muscular cell membrane and serum insulin-like growth factor-1 (IGF-1) in streptozotocin-induced diabetic rats. Methods: Rats in four experimental groups were investigated: the control group, the model group, the KYQWG group and the Metformin group. The insulin binding rate (IBR) of liver and skeletal muscular cell membrane was detected by receptor-ligand ra-diometric method and changes of serum levels of glucose, insulin and IGF-1 were observed before and after 4 weeks of medication. Results: The KYQWG group had a lower blood glucose level and ffiR of liver and muscular cell membrane, as compared with those in the model group (P<0. 01 or P<0.05), and a higher level of IGF-1 than that in the model group(P<0.01), but had no obvious changes in the serum level of insulin. Conclusion: KYQWG may increase the serum level of IGF-1 in diabetic rats, thus to decrease the insulin resistance a     

Intervention Effects of Jianpi Liqi Huoxue Decoction (健脾理气活血汤) on Lipid Peroxidative Liver Injury by Alcohol

Objective: To study the intervention effects of Jianpi Liqi Huoxue Decoction ( 健脾理气活血汤, JLHD) on lipid peroxidative liver injury induced by alcohol. Methods: The rat alcoholic model of liver disease (ALD) induced by Lieber-DeCarli liquid diet was established. Thirty-two male SD rats were randomly divided into 4 groups: the normal group ( n = 5), the control group ( n = 9), the model group ( n = 9) and the JLHD group (n = 9). From the 4th week after modeling, the rats were given JLHD or distilled water by gastrogavage respectively, and the samples of blood and liver tissues were taken out from the rats for determination by the end of the 8th week. The hepatic pathological changes were observed with HE staining; the liver injury related indices, including activity of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum, γ-glutamyl transpeptidase (γ-GT) activity and triglyceride (TG) content in liver tissues, as well as the lipid peroxidation related indices, including malonaldehyde (MDA) content and nitric oxide synthase (NOS) activity in liver tissue, serum Fe2 level, and the anti-peroxidation capacity related indices, including superoxide dismutase (SOD) activity, glutathion ( GSH) content and reactive oxygen species (antiROS) activity in liver tissues were determined. Results: ( 1 ) There were obvious figures of fatty degeneration and inflammatory infiltration in liver tissues of the model group. As compared with the control group, in the model group, the activity of ALT and AST, and Fe2 content in serum, γ-GT and NOS activity, TG and MDA content in liver tissues were significantly higher ( P＜0.01 ), while the activity of SOD, GSH and anti-ROS in liver tissues were significantly lower ( P＜0.01 ). (2) The fatty degeneration and inflammatory infiltration of liver tissues in the JLHD group were significantly lessen as compared with those in the model group; and the abnormalities of all the indexes revealed in the model rats were restored to certain extent in the JLHD group,and especially significant were the levels of ALT activity, MDA content and Fe2 , which were nearly normal.Conclusion: JLHD has significant effects against alcoholic liver injury, the acting mechanism of which is likely to be related with its anti-lipid peroxidative effect.     

Clinical significance of serum 7S collagen and type VI collagen levels for the diagnosis of hepatic fibrosis.

OBJECTIVE To measure serum 7S collagen (7S-C) and type VI collagen (VI-C) levels by radioimmunoassay (RIA) in Chinese patients with various liver disorders and in CCl4-treated SD rats, and to investigate the significance of the elevated levels of serum 7S-C and VI-C.

METHODS Serum 7S-C and VI-C levels were measured in 40 healthy control subjects, 168 patients with various liver disorders and non-hepatic diseases, and 52 CCl4-treated SD rats by using RIA which was developed in our hospital.

RESULTS Serum 7S-C and VI-C were significantly elevated in patients with chronic active hepatitis (CAH), liver cirrhosis (LC), hepatic cellular carcinoma (HCC) (P < 0.01 respectively), chronic persistent hepatitis (CPH), and some with non-hepatic diseases (P < 0.05). Serum 7S-C, serum laminin and hyaluronic acid were well correlated. Serum 7S-C and VI-C were not closely correlated. Both collagens were correlated with serum albumin/globulin ratio, aminotransferase and total bilirubin, not with alkaline phosphatase. In CCl4-treated SD rats, serum 7S collagen and type VI collagen levels were correlated with the degree of hepatic fibrosis.

CONCLUSIONS Serum 7S collagen and type VI collagen are useful markers for diagnosing liver fibrosis. And the combined measurement of IV-C, VI-C and other markers of connective tissue metabolism or biochemical data seems to provide additional information to predict progressive hepatic fibrosis.

     

Intervention effects of Jianpi Liqi Huoxue Decoction (健脾理气活血汤) on lipid peroxidative liver injury induced by alcohol

Objective： To study the intervention effects of Jianpi Liqi Huoxue Decoction （健脾理气活血汤, JLHD） on lipid peroxidative liver injury induced by alcohol. Methods： The rat alcoholic model of liver disease （ALD） induced by Lieber-DeCarli liquid diet was established. Thirty-two male SD rats were randomly divided into 4 groups ： the normal group （ n = 5）, the control group （ n = 9）, the model group （ n = 9） and the JLHD group （ n =9）. From the 4th week after modeling, the rats were given JLHD or distilled water by gastrogavage respectively, and the samples of blood and liver tissues were taken out from the rats for determination by the end of the 8th week. The hepatic pathological changes were observed with HE staining; the liver injury related indices, including activity of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in serum, γ-glutamyl transpeptidase （γ-GT） activity and triglyceride （TG） content in liver tissues, as well as the lipid peroxidation related indices, including malonaldehyde （MDA） content and nitric oxide synthase （NOS） activity in liver tissue, serum Fe^2＋ level, and the anti-peroxidation capacity related indices, including superoxide dismutase （SOD） activity, glutathion （GSH） content and reactive oxygen species （anti- ROS） activity in liver tissues were determined. Results： （ 1 ） There were obvious figures of fatty degeneration and inflammatory infiltration in liver tissues of the model group. As compared with the control group, in the model group, the activity of ALT and AST, and Fe^2＋ content in serum, γ-GT and NOS activity, TG and MDA content in liver tissues were significantly higher （ P〈0. 01 ）, while the activity of SOD, GSH and anti-ROS in liver tissues were significantly lower （P〈0.01）. （2） The fatty degeneration and inflammatory infiltration of liver tissues in the JLHD group were significantly lessen as compared with those in the model group; and the abnormalities of all the indexes revealed in the model rats were restored to certain extent in the JLHD group, and especially significant were the levels of ALT activity, MDA content and Fe^2＋ , which were nearly normal. Conclusion： JLHD has significant effects against alcoholic liver injury, the acting mechanism of which is likely to be related with its anti-lipid peroxidative effect.     

Telmisartan protects against insulin resistance by attenuating inflammatory response in rats

This study investigated the effects of telmisartan on insulin resistance in high-fat diet-treated rats and the possible mechanism.A total of 40 male Sprague-Dawley rats enrolled in the study were divided into 4 groups at random:ND group(n=10) and HD group(n=10),in which the rats were given a normal chow diet or a high-fat diet for 20 weeks following a one-week adaptation;ND+telmisartan(n=10) group and HD+telmisartan group(n=10),in which the rats were initially administered in the same way as the ND or HD group,and then they were orally gavaged with telmisartan(5 mg/kg daily) additionally for 5 weeks.Related inflammatory factors were measured by ELISA.Monocyte chemotactic protein 1(MCP-1),phosphorylated JNK and IκB-α expressions in both adipose and liver were detected by Western blotting.CRP and angiotensin Ⅱ receptor 1(AT1) mRNA expressions in both adipose and liver were determined by RT-PCR.The results showed that telmisartan administration in vivo reversed insulin resistance as evidenced by a decrease in plasma fasting glucose levels,plasma fasting insulin levels and homeostasis model of assessment-insulin resistance(HOMA-IR).Furthermore,telmisartan administration significantly reduced serum CRP,TNF-α and IL-1β levels,and elevated serum IL-10 levels.It was also found to hamper the high-fat diet-induced increase in CRP mRNA,AT1 mRNA and MCP-1,and decrease in IκB-α in both adipose and liver.It was concluded that telmisartan administration in vivo may improve insulin resistance through attenuated inflammatory response pathways.     

Effects of isoflurane on ICAM-1 expression and neutrophils infiltration in rats with liver ischemia and reperfusion injury

Objective： To establish a rat model of warm partial hepatic ischemia-reperfusion （IR）, and investigate the protective and anti-inflammatory effects of isoflurane on warm hepatic ischemia-reperfusion injury （IRI） in rats. Methods： Thirty-two female Sprague-Dawley rats were divided equally into 4 groups （n-8）： PB-Sham group in which the rats were anesthetized by intraperitoneal injection of pentobarbital sodium （1.0%, 40 mg/kg, PB） and received a sham operation without occlusion of liver blood flow; PB-IR group whose rats underwent partial hepatic IR after anesthesia; Iso-Sham group in which inhalation of 1.0 MAC isoflurane and sham operation was performed; Iso-IR group in which 1.0 MAC isoflurane was inhaled for 4 h and IR was performed. Rat model of warm partial hepatic IR was established by clamping the hepatic arteries and hilar vessels distributing to the left and median lobes to induce partial hepatic ischemia （70%） for 60 rain followed by reperfusion for 3 h. The rats were killed 3 h after declamping, and specimens of liver tissue and blood were obtained. The serum ALT and AST were detected as liver damage markers. Viability of myeloperoxidase （MPO） in liver was measured. The protein level of ICAM-1 in the liver was detected by immunohistochemistry and Western blotting. Results： Rats treated with 1.0 MAC isoflurane during warm partial （70%） hepatic ischemia 60 rain and 3 h reperfusion had significantly lower serum ALT and AST compared with rats anesthetized with pentobarbital sodium subjected to hepatic IRI. The expression of ICAM-1 in hepatic tissue was significantly increased by hepatic IRI after pentobarbital sodium anesthesia. Isoflurane significantly inhibited protein expression of ICAM-1 in hepatic IR injury compared with pentobarbital sodium anesthesia. Viability of liver MPO was significantly increased by hepatic IRI after pentobarbital sodium anesthesia; Isoflurane can significantly inhibit MPO alteration in rat liver ischemia-reperfusion injury compared with rats anesthetized with pentobarbital sodium. Conclusion： Isoflurane anesthesia can attenuate liver IR injury in rats that maybe by inhibiting ICAM-I expression and reducing the infiltration of neutrophils.     

Effects of Acupuncture on Progesterone and Prolactin in Rats of Embryo Implantation Dysfunction

Objective:To investigate the effect of acupuncture on progesterone(P4) and prolactin(PRL) in rats of embryo implantation dysfunction(EID).Methods:On the first day of pregnancy,72 female Wistar rats were randomly allocated into the normal group,the EID model group,the acupuncture group and the P4 group(18 in each group).The normal group was injected sesame oil,while the other three groups were given mifepristone to establish the EID model.The acupuncture group and the P4 group were given treatment of acupuncture and P4 injection,respectively.The serum of P4 and PRL were detected by radioimmunoassay,and the mRNA and protein expressions of P4 receptor(PR) and PRL receptor(PRLR) were detected using real-time polymerase chain reaction and immunohistochemical method,respectively.Results:Compared with the normal group,the serum levels of P4 and PRL as well as the mRNA and protein expression levels of PR and PRLR in the EID model group were significantly lowered(P<0.01 or P<0.05).The above indices in the acupuncture group and the P4 group were significantly elevated compared with the EID model group(P<0.01 or P<0.05).Conclusion:Acupuncture can promote embryo implantation effectively,which might be related to the effects of acupuncture on upregulating the P4 and PRL levels in serum and the PR and PRLR expression levels in rats.     

Study on effects of Zhi Zi(Fructus gardeniae) on non-alcoholic fatty liver disease in the rat

OBJECTIVE:To investigate the effects of Zhi Zi(Fructus Gardeniae) on non-alcoholic fatty liver disease(NAFLD) induced by a high-fat diet in the rat.METHODS:A rat model of NAFLD was established using a high-fat diet.Twenty one rats were randomly divided into a normal group,a model group and a Zhi Zi treatment group,7 rats per group.Drinking water and the drug were intragastrically administrated for 5 weeks.Samples were then taken to observe pathological changes of the liver tissue(HE staining);changes in the fat metabolism pathway e.g.triglyceride(TG) and free fatty acid(FFA) content;alterations in liver function,i.e.serum alanine aminotransferase(ALT) and aspartate aminotransferase(AST) activity;and differences in tumor necrosis factor α(TNF-α) and P-IkB protein expression in the liver tissue.RESULTS:Fatty degeneration and vacuole-like changes of different degrees occurred in hepatic cells of the model group.Markers for fat metabolism,serum ALT and AST activities,and expressionof TNF-α and P-IkB proteins in liver tissue significantly increased.Fat metabolism in the Zhi Zi group significantly reduced,as shown by a drop in marker levels.Serum ALT and AST activities,and expression of TNF-α,P-IkB proteins in liver tissue were also significantly decreased in this group.CONCLUSION:Zhi Zi has a very strong inhibitory action on lipidosis and inflammatory injury in the rat model of NAFLD.This mechanism may possibly be related to the inhibition of the free fatty acid metabolism pathway.     

Multiple Pharmacological Actions of Yiqi Huatan Decoction (益气化痰方) in A Model of Depression in Rats

Objective: To investigate the influence of Yiqi Huatan Decoction (益气化痰方, YHD) on a model of depression in rats under different pathological conditions. Methods: Thirty-two male SD rats were randomly divided into 4 groups of 8: normal, model, YHD, and maprotiline. The model group, YHD group and maprotiline group used separate feeding and rats were exposed to chronic and unpredictable stress to build the depression model. From day 2, the YHD group and maprotiline group were respectively given YHD (7 g/kg) and maprotiline (10 mg/kg) by gastrogavage once daily. The normal and model groups were given the same volume of drinking water. The medication duration were 21 days. At the end of the experiment, the serum levels of copper and zinc were determined by atomic absorption spectroscopy, plasma concentrations of adrenocorticotropic hormone (ACTH) and cortisol (COR) were detected by radioimmunoassay, and levels of norepinephrine (NE), dopamine (DA), and 5-hydroxytryptamine (5-HT) in the hypothalamus were analysed by high performance liquid chromatography-eletricochemistry. Results: Compared with the content of copper and zinc in the serum of rats in the normal group, serum copper levels in model rats were significantly increased and zinc content was significantly reduced (both P<0.05). Plasma concentrations of ACTH and COR in the model group were significantly increased compared with those in the normal group (P<0.05, P<0.01). The contents of NE, DA, and 5-HT in the hypothalamus of rats in the model group were significantly reduced compared with those of the normal group (P<0.05 or P<0.01). Compared with those in the model group, the serum copper content and plasma concentrations of ACTH and COR were significantly decreased (all P<0.05); meanwhile, serum zinc content and hypothalamic contents of NE, DA, and 5-HT were significantly increased in rats of the YHD group (all P<0.05). The same effects were also shown in the maprotiline group except for 5-HT (all P<0.05). Conclusion: The pharmacological actions of YHD for depression might be related to improving trace-element anomalies, reversing endocrine dysfunction, and modulating the disorders of monoaminergic neurotransmitters.     

Effects on the Tumor Specific Growth Factor and Tumor Necrosis Factor α in Rats' Precancerous Lesion of Primary Hepatocellular Carcinoma by Direct Moxibustion at Ganshu(BL 18) Acupoint

Objective: To investigate the effect of direct moxibustion at Ganshu(BL18) on the serum concentrations of tumor specific growth factor(TSGF) and tumor necrosis factor α(TNF-α) in a rat model with precancerous lesion of primary hepatocellular carcinoma(HCC), so as to explore the mechanism of moxibustion underlying improvement of HCC. Methods: Sixty male Wistar rats were randomly divided into control group(n=10), model group(n=20), prevention group 1(n=15) and prevention group 2(n=15). The normal rats were injected with physiological saline as blank control. At the same time, the rats of other three groups were injected with diethylnitrosamine to establish the HCC model. Direct moxibustion with grain-sized moxa was applied to bilateral Ganshu acupoint of the rats in the prevention group 1(1 treatment course, 20 days) and prevention group 2(2 treatment courses, 40 days), 5 doses for each acupoint, 0.5 mg/dose, once every other day. At each time point(before model establishment, the end of 1st course prevention, the end of 2nd course prevention and the end of model establishment), serum levels of TSGF and TNF-α were detected using enzyme-linked immunosorbent assay. Results: Compared with the control group, there was a remarkably increase of serum TSGF and TNF-α contents in the model group at the end of the experiment(P0.05). At the end of the 1st course of direct moxibustion, the contents of serum TSGF and TNF-α of rats in the prevention group 1 were significantly increased compared with that of the model group(P0.05). At the end of the 2nd course of direct moxibustion, serum TSGF and TNF-α levels of rats in the model group were higher than the normal group with significantly difference(P0.05), and the levels of TSGF and TNF-α in the prevention group 2 were significantly reduced in comparison with the model group(P0.05). Conclusion: It was possible that direct moxibustion could inhibit precancerous lesion and postpone hepatocarcinogenesis, and the therapeutic effect of two courses were better than one course.     

Antifibrotic Effect of Total Flavonoids of Astmgali Radix on Dimethylnitrosamine-Induced Liver Cirrhosis in Rats

Objective: To study the effect of total flavonoids of Astmgali Radix (TFA) on liver cirrhosis induced with dimethylnitrosamine (DMN) in rats, and the effect on peroxisome proliferator-activated receptor γ (PPARγ), uncoupling protein 2 (UCP2) and farnesoid X receptor (FXR). Methods: Fifty-three Sprague-Dawley rats were randomly divided into a control group (10 rats) and a DMN group (43 rats). Rats in the DMN group were given DMN for 4 weeks and divided randomly into a model group (14 rats), a low-dosage TFA group (14 rats) and a high-dosage TFA group (15 rats) in the 3rd week. Rats were given TFA for 4 weeks at the dosage of 15 and 30 mg/kg in the low- and high-TFA groups, respectively. At the end of the experiment blood and liver samples were collected. Serum liver function and liver tissue hydroxyproline content were determined. hematoxylin-eosin (HE), Sirus red and immunohistochemical stainings of collagen Ⅰ, smooth muscle actin (α-SMA) was conducted in paraffinembedded liver tissue slices. Real time polymerase chain reaction (PCR) was adopted to determine PPARγ, UCP2 and FXR mRNA levels. Western blot was adopted to determine protein levels of collagen Ⅰ, α-SMA, PPARγ, UCP2 and FXR. Results: Compared with the model group, TFA increased the ratio of liver/body weight (low-TFA group P<0.05, high-TFA group P<0.01), improved liver biochemical indices (P<0.01 for ALT, AST, GGT in both groups, P<0.05 for albumin and TBil in the high-TFA group) and reduced liver tissue hydroxproline content (P<0.01 in both groups) in treatment groups significantly. HE staining showed that TFA alleviated liver pathological changes markedly and Sirus red staining showed that TFA reduced collagen deposition, alleviated formation and extent of liver pseudolobule. Collagen Ⅰ and α-SMA immunohistochemical staining showed that staining area and extent markedly decreased in TFA groups compared with the model group. TFA could increase PPARγ, it regulated target UCP2, and FXR levels significantly compared with the model group (in the low-TFA group all P<0.05, in the high group all P<0.01). Conclusions: TFA could improve liver function, alleviate liver pathological changes, and reduce collagen deposition and formation of liver pseudolobule in rats with liver cirrhosis. The antifibrotic effect of TFA was through regulating PPARγ signal pathway and the interaction with FXR.     

Antifibrotic Effect of Total Flavonoids of Astmgali Radix on Dimethylnitrosamine-lnduced Liver Cirrhosis in Rats

Objective:To study the effect of total flavonoids of Astmgali Radix(TFA) on liver cirrhosis induced with dimethylnitrosamine(DMN) in rats,and the effect on peroxisome proliferator-activated receptor 7(PPAR 7),uncoupling protein 2(UCP2) and farnesoid X receptor(FXR).Methods:Fifty-three Sprague-Dawley rats were randomly divided into a control group(10 rats) and a DMN group(43 rats).Rats in the DMN group were given DMN for 4 weeks and divided randomly into a model group(14 rats),a low-dosage TFA group(14 rats) and a high-dosage TFA group(15 rats) in the 3rd week.Rats were given TFA for 4 weeks at the dosage of 15 and 30 mg/kg in the low- and high-TFA groups,respectively.At the end of the experiment blood and liver samples were collected.Serum liver function and liver tissue hydroxyproline content were determined,hematoxylin-eosin(HE),Sirus red and immunohistochemical stainings of collagen Ⅰ,smooth muscle actin(α-SMA) was conducted in paraffinembedded liver tissue slices.Real time polymerase chain reaction(PCR) was adopted to determine PPAR 7,UCP2 and FXR mRNA levels.Western blot was adopted to determine protein levels of collagen Ⅰ,α-SMA,PPAR γ,UCP2 and FXR.Results:Compared with the model group,TFA increased the ratio of liver/body weight(low-TFA group P0.05,high-TFA group P0.01),improved liver biochemical indices(P0.01 for ALT,AST,GGT in both groups,P0.05 for albumin and TBil in the high-TFA group) and reduced liver tissue hydroxproline content(P0.01 in both groups) in treatment groups significantly.HE staining showed that TFA alleviated liver pathological changes markedly and Sirus red staining showed that TFA reduced collagen deposition,alleviated formation and extent of liver pseudolobule.Collagen Ⅰ and α-SMA immunohistochemical staining showed that staining area and extent markedly decreased in TFA groups compared with the model group.TFA could increase PPAR γ,it regulated target UCP2,and FXR levels significantly compared with the model group(in the low-TFA group all P0.05,in the high group all P0.01).Conclusions:TFA could improve liver function,alleviate liver pathological changes,and reduce collagen deposition and formation of liver pseudolobule in rats with liver cirrhosis.The antifibrotic effect of TFA was through regulating PPAR γ signal pathway and the interaction with FXR.