OBJECTIVE:To observe the effects of different therapeutic methods and the recipes of Chinese medicine (CM) on the activation of c-Jun N-terminal kinase (JNK) in Kupffer cells of rats with fatty liver disease and to explore the mechanisms of these therapeutic methods.METHODS:By using a random number table, 98 rats were randomly divided into 7 groups: control group, model group, and 5 treatment groups, including soothing Liver (Gan) recipe group, invigorating Spleen (Pi) recipe group, dispelling dampness recipe group, promoting blood recipe group, and complex recipe group. Rats in the control group were fed with normal food and distilled water by gastric perfusion, while rats in the model group were fed with high-fat food and distilled spirits by gastric perfusion. Rats in the 5 treatment groups were fed with high-fat food and corresponding recipes by gastric perfusion. Twelve weeks later, all rats were sacrificed and liver tissues were stained for pathohistological observation. Kupffer cells were isolated from livers of rats to evaluate JNK and phospho-JNK expressions by Western blotting.RESULTS:The grade of hepatic steatosis was higher in the model group than the control group (P<0.05). Compared with the model group, the grade of fatty degeneration in soothing Liver recipe group and invigorating Spleen recipe group were significantly ameliorated (P<0.05). Expressions of JNK and phospho-JNK in Kupffer cells were significantly higher in the model group than those in the control group (P<0.05, P<0.01). Compared with the model group, expressions of JNK in all treatment groups decreased, especially in invigorating Spleen recipe group and promoting blood recipe group (P<0.05). Compared with the model group, expressions of phospho-JNK in all treatment groups declined significantly (P<0.01), especially in soothing Live recipe group and invigorating Spleen recipe group.CONCLUSIONS:The high expressions of JNK and phospho-JNK in Kupffer cells might play an important role in the pathogenesis of fatty liver disease in rats. The recipes of CM, especially invigorating Spleen recipe and soothing Liver recipe, might protect liver against injury by reducing the total JNK protein content and inhibiting the activation of JNK protein in Kupffer cells of fatty liver model rats, which showed beneficial effects on fatty liver disease.

AIM: To study the effects of panax notoginseng saponins(PNS) on the content of triglyceride(TG) and the mRNA expression of liver X receptor α(LXRα) in steatotic hepatocyte L02.METHODS: The cells of hepatocyte L02 were cultured with 50% bovine calf serum for 48 h and a model of steatosis of hepatocytes was established.The appropriate concentrations of PNS on steatotic hepatocytes were detected by MTT assay.The cells were divided into 5 groups: normal control group,model group,spontaneous recovery group,low concentration of PNS group(10 mg/L) and high concentration of PNS group(50 mg/L).Meanwhile,hepatocytes in model group were continuously cultured by RPMI-1640 medium containing 50% bovine calf serum while others were cultured by RPMI-1640 medium containing 10% bovine calf serum.After 24 h,the lipid droplets in the cells stained with oil red O were observed under microscope,the intracellular TG levels were determined by automatic biochemical analyzer and the mRNA expression of LXRα in hepatocytes was examined by RT-PCR.RESULTS: Compared to normal control group,Oil red O staining presented numerous orange-red or red lipid droplets in the cytoplasm of human L02 hepatocytes in model group.The content of TG in model group were significantly increased(P0.01).After treated with PNS for 24 h,the content of TG in PNS treatment groups was significant decreased than that in spontaneous recovery group(P0.05),especially in low concentration of PNS group(P0.01).The accumulation of lipid droplets in low concentration of PNS group was decreased significantly.Compared to normal control group,the mRNA expression of LXRα in model group was significantly up-regulated(P0.01).Compared to spontaneous recovery group,the mRNA expression of LXRα in PNS treatment groups declined in different levels,especially in low concentration group(P0.05).CONCLUSION: The deposition of lipid in hepatocytes might be related to the high expression of LXRα mRNA.PNS decreases the content of TG in steatotic hepatocytes and promotes the recovery of hepatocyte steatosis by downregulating the mRNA level of LXRα.

目的观察三七总皂甙(PNS)对脂肪变性肝细胞的降脂作用及机制。方法采用50%小牛血清诱导L02肝细胞48 h建立L02肝细胞脂肪变性模型,分为模型组、自然恢复组、PNS低剂量组、PNS高剂量组,并设正常组。除模型组继续予含50%小牛血清的1640培养基培养外,余组均改予含10%小牛血清培养。PNS低、高剂量组分别予10、50μg/ml PNS作用。药物作用24 h后,油红O染色观察肝细胞内脂滴变化,全自动生化仪检测TG水平,RT-PCR法检测固醇元件结合蛋白-1c(SREBP-1c mRNA)表达。结果与自然恢复组比较,PNS各治疗组细胞内脂滴少于其他组,低剂量组更为明显;TG水平明显低于其他各组(P<0.05),低剂量组下降更为显著(P<0.01);SREBP-1c mRNA的表达量均有下降,低剂量组更为明显(P<0.05)。结论PNS能显著降低脂肪变性肝细胞内TG水平,减轻肝细胞脂肪变性;机制可能与下调SREBP-1c mRNA的表达有关。

Objective To observe the effect of soothing liver or invigorating spleen recipes of Traditional Chinese Medicine(TCM) on expression of PI3K p85α protein in hepatocyte of rats with non-alcoholic fatty liver disease(NAFLD),and to explore their mechanisms of it.Methods The rat models of NAFLD were established by feeding on high fat diet for 12 weeks.Then the model rats were randomly subdivided into 5 subgroups: model group,soothing liver group,invigorating spleen group,integrated group and natural recovery group.Rats in medication groups were fed with corresponding herbs by gastric perfusion while the remaining rats were fed by equal distilled water.Meantime,rats in model group were continuously given high fat diet while all other rats were given normal diet.8 weeks later,all rats were anesthetized by intraperitoneal injection of 3% pentobarbital.ALT and AST contents in the serum and together with blood lipid were detected by automatic biochemical analyzer.The degree of insulin resistant was measured by homeostasis model assessment.Hepatic tissue was stained to undergo pathohistology observation.The expression of PI3K p85α protein in hepatocyte was measured by immunohistochemistry.Results Compared with model group,the hepatic steatosis degree were lighten obviously in medication groups and natural recovery group,the levels of ALT,AST,blood lipid and insulin resistance were ameliorated significantly(respectively,P<0.01;P<0.05),expressions of PI3K p85α protein in hepatocyte were reduced significantly(P<0.05).Compared with the natural recovery group,the levels of ALT,AST,blood lipid in medication groups were shown no statistical significance,but hepatic steatosis degree were lighten,insulin resistance index and expressions of PI3K p85α protein in hepatocyte were reduced significantly(P<0.05).Conclusion Soothing liver,invigorating spleen methods and recipes showed certain therapeutic effect on rats with NAFLD,which may be due to its effects in decreasing expression of PI3K p85α protein in hepatocyte.

Objective To observe the effect of Jiangzhi Ninggan capsule on acute hepatic injury induced by D-galactosamine(D-Gal) in mice.Methods Sixty mice were divided into six groups at random: model group,control group,Bifendate group(positive control group),Jiangzhi Ninggan Capsule groups(high dose,middle dose,low dose).The mice in treatment groups were administered medicine respectively via gavage,while the others in control and model groups were treated with the same dose of nomal saline.After six days,all of the mice except those in the control group were treated with intraperitoneal injection of 10% D-Gal(1 000 mg/kg).twenty-four hour later,the mice were killed and the blood sample and liver tissue were taken.Liver pathological changes were observed and the activities of serum ALT,AST,and the content of liver MDA were detected.Results Compared with the model group,the effect on levels of ALT,AST in blood and the content of MDA in liver tissue of Jiangzhi Ninggan Capsule groups(high dose、middle dose) were reduced significantly(respectively,P0.05;P0.01;P0.001),and the liver pathological changes were lightened obviously in dose.Conclusion Jiangzhi Ninggan capsule has protective acute hepatic injury induced by D-galactosamine.

Objective To investigate the effect of different therapies on hepatic nuclear factor κBp65(NF-κBp65) and Kuffer cell p38MAPK expression in rats with fatty liver.Methods Ninety-eight SD rats were randomized into 7 groups: normal group,model group,liver-soothing group(receiving gavage of Chaihu Shugan Powder 3.2 g·kg-1·d-1),spleen-strengthening group(receiving gavage of Shen Ling Baizhu Powder 10.0 g·kg-1·d-1),dampness-respelling group(receiving gavage of Pingwei Powder 3.5g·kg-1·d-1),blood-activating group(receiving gavage of Gexia Zhuyu Decoction 7.1 g·kg-1·d-1) and the combination group(receiving gavage of selt-made prescription 11.9 g·kg-1·d-1).Except the normal group,the rats in other groups were fed with high-fat forage and wine to induce NAFLD.After treatment for 12 weeks,the rats were executed to obtain the liver.Immunohistochemical assay was used for the detection of hepatic NF-κBp65 activity and hepatic pathological changes.Kupffer cells were isolated from 3 rats of each group,and then the p38MAPK protein expression and phosphorylation p38MAPK level in Kupffer cells were detected by Western blot method.Results Hepatic NF-κBp65 protein expression was increased in the model group(P0.05 compared with the model group),the incidence of NF-κBp65 positive expression was the lowest and hepatic fatty changes were the slightest in liver-soothing group and spleen-strengthening group(P0.05 compared with the model group).The increase of hepatic NF-κBp65 expression in NAFLD rats was positively correlated with the hepatic pathological changes(P0.01).Kupffer cell p38MAPK protein expression and phosphorylation p38MAPK protein expression were increased in the model group(P0.01 compared with the normal group),while were decreased in the medication groups(P0.01 compared with the model group),in particular in spleen-strengthening group and liver-soothing group.Conclusion Inhibition of hepatic NF-κBp65 expression,and Kupffer cell p38MAPK protein expression and phosphorylation p38MAPK protein expression may be one of their therapeutic mechanisms of the above therapies for NAFLD.

Objective:To detect the function of Jiangzhi Ninggan capsule in enhancing the angiectasia of the capillaries and improving the microcirculation in mice's ala auris,and thus to explore the effects of Jiangzhi Ninggan capsule on the microcirculation of the mice's ala auris.Methods:The method we used here was administering the drug directly.Results:The method we used here was adminnistering the drug directly.Results:Jiangzhi Ninggan capslue could expand signinficantly the diameter of micro-artery,the diameter of micro-vein and improve the microcirculation in normal mice auricular(P0.01,P0.001).Conclusion:The fact that Jiangzhi Ninggan capsule could enhance the angiectasia of the capillaries and improve the microcirculation has provided the ecidences for its application in clinical medicine.

Objectives: To observe the relation of serum supersensitivity C reactive protein and different syndromes of cerebral infarction,and the degree of nervous system defect of different syndromes of cerebral infarction,and to analyse the relations.Offer the gist of clinical diagnosis and syndrome differentiation of TCM of cerebral infarction.Methods: As for 82cases of cerebral infarction,the different syndromes were differed,serum hs-CRP,serum TG,TC and HDL-C were detected.The value of TC/HDL was calculated,and the degree of nervous system defect was evaluated.The relation of different syndromes of TCM was analysed.Results: The hs-CRP is manifest abnormality and the degree of nervous system defect are manifest in the patients with wind-phlegm stasis,qi deficiency with blood stasis,and have significant deviation with those in patients with yin deficiency and liver wind.The values of TC,TG and TC/HDL have no significant deviation in the three syndromes.The value of hs-CRP has positive correlation with that of TG,TC and TC/HDL.The value of hs-CRP has correlation with the degree of patients with nervous system defect.Conclusions: The rise of the value of hs-CRP is the important index of phlegm syndrome,blood stasis of cerebral infarction.The rise of the value of hs-CRP maybe become one of the index of appraisal of patient's condition.

Objective: To observe the therapeutical effect of Jiangzhi Ninggan capsule on rats with fatty liver disease,and to explore the mechanism of it.Methods: The rat models with fatty liver disease were established by feeding on high fat diet combined with ethanol via gavage for 6 weeks.Then the rats were random divided into seven groups:Jiangzhi Ninggan capsule groups(high dose,middle dose,low dose),Dongbao Gantai pill group,natural recovery group,model group and control group.4 weeks later,Liver pathologic changes were observed together with the activity of serum ALT,AST,the blood lipid,and the content of liver MDA,GSH,TG,CHOL.Results: Compared with the natural recovery group,the level of ALT,CHOL and TG in blood of Jiangzhi Ninggan capsule groups were reduced significantly(respectively,P0.01;P0.05),and the content of CHOL,TG,MDA,GSH in liver tissue were ameliorated(respectively,P0.01;P0.05),furthermore it show dose-effect relation,the effect of high dose group was correspond with Dongbao Gantai pill group;The hepatic steatosis degree were lighten obviously(P0.01).Conclusion: Jiangzhi Ninggan capsule had good effect on fatty liver disease.This may be due to its effects in decreasing blood lipid and prevention of lipid peroxidation.

探讨中医正气学说与现代免疫学理论的相关性,提出从中医免疫观防治慢性乙型肝炎(CHB)的观点,并对单味中药或中药单体调整机体内部的细胞免疫和体液免疫环节、有效结合机体免疫系统的微观作用与中医辨证思雏的宏观作用作了归纳,还对中药复方的免疫调节作用、对免疫耐受状态的消除作用、免疫双向调节作用作了总结和分析。从中医免疫观的角度出发,结合现代医学对中医药的研究进展,指出中医药在防治CHB方面的作用及优势,为CHB的临床治疗提供新的思路。