| Objective: Herpetospermum seed is a Traditional Tibetan medicine for the prevention and treatment of liver disease,can clear away heat and toxic material,also protect liver and gallbladder.Herpetospermum seed mainly used for the protection of hepatitis clinically,and liver disease adjuvant therapy.The experimental results showed that the total lignans of Herpetospermum seed were the effective parts of the treatment of liver disease and the liver injury caused by CCl4(carbon tetrachloride),alcohol and so on.And it has a definite protective effect with the immune liver injury caused by Con A.Due to physical,chemical,drug or other factors in the cause of liver injury,the body itself will secrete a variety of factors self-regulation to repair liver function and damage,while stimulate the regeneration of liver cells to a certain extent.Whether the total lignans of Herpetospermum seed a repair effect on liver injury caused by 70 %(2/3)of liver removal and whether it has been shown to promote liver regeneration is not yet clear.Therefore,this study will establish a 70 %(2/3)hepatectomy model of mice.The effect and mechanism of the total lignans from the total lignans on the liver function,liver regeneration.The effect of the total lignans on the liver resection and the effect of liver regeneration on liver regeneration were studied in this paper.Methods: 1.Mouse 70 %(2/3)hepatectomy model was established: A model of liver resection was established by ligation resection,and mice were anesthetized by ether under sterile conditions,and the liver was exposed to the abdomen of mice(About 68 % of the weight of the original liver of the mice)Sutured the abdomen and waited for the mice to wake up.Recording their weight and liver weight.Successful wake of mice and freedom of movement as a sign of successful surgery.2.The effects of total lignans of Herpetospermum seed the liver function and liver regeneration after 70 %(2/3)hepatectomy in mice:(1)The effect of the total lignans of Herpetospermum seed the liver repair of 70 %(2/3)hepatectomy in mice: Mice were divided into blank group,model(surgery)group,salvia miltiorrhiza injection group,total lignans(15 g/kg)group and total lignans(25 g/kg)group.(1)The serum levels of AST,ALT and ALB in hepatectomized mice were detected by biochemical method and the levels of CRP were detected.The expression of PCNA and Ki67 protein in liver tissue of hepatectomy mice were detected by histopathological examination.The liver regeneration rate was calculated and the liver regeneration rate was calculated.(2)To detect the histopathology.The expression of PCNA and Ki67 protein in hepatocytes were detected by immunohistochemistry.Calculate HRR.To observe the Effect of total lignans of Tibetan medicinal herpetospermum seed on liver regeneration after hepatectomy in mice.(2)The effect of the total lignans of Herpetospermum seed with the liver repair of 70 %(2/3)hepatectomy in mice of different age: Mice were divided into 4 groups according to age,low age group,low age total lignans 25 g/kg group,the elderly model group,and the elderly total lignans 25 g/kg group.To establish the model of 70 %(2/3)hepatectomy in mice.(1)Biochemical detection of liver resection of mice serum AST,ALT and ALB were observed to observe the effect of the drug on the liver function of mice in different age.(2)The expression of PCNA protein in hepatocytes of hepatectomy mice was detected by immunohistochemical method.To observe the effect of drugs on postoperative liver regeneration in mice of different age.3.The effects of total lignans on the liver repair of 70 %(2/3)hepatectomy in mice: A 70 %(2/3)hepatectomy model of mice was established.The levels of IL-6,NF-κB and TNF-a in liver tissue of hepatectomized mice were detected,and the mechanism of repairing liver in mice was investigated.Results: 1.Mouse 70%(2/3)hepatectomy model was established: From anesthetized mice to suturing and disinfecting the incision to complete a mouse operation for 15-20 min.The success rate of surgery in mice was 93.4 % under the premise of ensuring the survival of each group.Successful mice can be free after surgery,normal diet,and abdominal wounds healed well.2.The effects of total lignans on the liver function and liver regeneration after 70%(2/3)hepatectomy in mice:(1)The effect of the total lignans of Herpetospermum seed the liver repair of 70 %(2/3)hepatectomy in mice: Compared with the blank group,the AST activity of the model group mice increased,the activity of ALT and the content of CRP reached the peak at about 7 days.The activity of the enzyme was lower than that of the control group.Compared with the model group,the levels of ALT and AST in total lignans group and salvia miltiorrhiza injection groups were decreased and the levels of CRP were decreased.The pathological changes of liver tissues were observed.The necrosis,edema and edema of mice liver cells were observed in the short time inflammation.Compared with the model group,the corresponding drug-induced mouse cells to improve the situation,and the number of liver cell proliferation gradually increased.The cell morphology also changes with time tend to normal.Observed in mice after PCNA and Ki67 expression,and blank Compared with the model group,the expression of PCNA and Ki67 protein in the mice were increased in different stages.(2)The effect of total lignans on the liver repair of 70 %(2/3)hepatectomy in different age mice: The AST and ALT activities of mice in different months after hepatectomy and the contents of ALB in the mice were significantly lower than those in the model group(P <0.05).Compared with the model group,the contents of ALB were increased.The histopathological changes of the mice in different age groups were observed.The mice in the model group showed a large amount of edema,vacuolar degeneration and necrosis at 1 d.Compared with the model group,and the number of hepatocytes in the total lignans group was different.The vacuolar vacuoles were changed or the degeneration of the granules,and the cell division of the hepatocytes increased,then the cell volume became smaller.The expression of PCNA in mouse mice was significantly higher than that in the control group.Compared with the model group,the expression of PCNA protein in mice of different age of lignans was increased in different degrees,but there was no statistically significant difference between mice in different months.3.The effects of total lignans on the liver repair after 70 %(2/3)hepatectomy in mice: Compared with the blank group,the levels of IL-6,NF-κB and TNF-a in the model group were higher than those in the control group.Compared with the model group, the total lignans could decrease the levels of IL-6,NF-κB and TNF-a in the early stage,suggesting that the mechanism IL-6/STAT-3 and other signal pathways related to Tibetan medicine Herpetospermum seed total lignans can reduce the injury caused by surgery and inflammation,and protect the liver.Conclusion: The above experimental results show that the total lignans of Herpetospermum seed have a certain protective effect on 70 % hepatectomy mice,which can promote the liver regeneration and prolong the regeneration and repair time.IL-6/STAT-3 and other signaling pathways have different anti-inflammatory and anti-inflammatory effects on liver regeneration in different age,and it is speculated that the mechanism may be related to the release of inflammatory factors,and the reduction of inflammation and the recovery of cell morphology. |
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