BJ-JN抗肝纤维化作用及其机理的研究
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摘要
利用D-半乳糖胺(D-GalN)、四氯化碳(CCl_4)、卡介苗加脂多糖(BCG+LPS)分别诱导小鼠急性肝损伤、大鼠肝纤维化与小鼠免疫性肝损伤模型，研究了BJ-JN(3，6，12g生药·kg~(-1)·d~(-1))对上述不同原因所诱导肝损伤的防治作用及其作用机理。并且用血清药理学方法研究了BJ-JN含药血清体外对肝星状细胞(HSC)功能的影响。主要结果概括如下：
1．BJ-JN对D-GalN诱导小鼠急性肝损伤的保护作用
BJ-JN高剂量组可明显降低D-GalN升高的小鼠肝脏指数和血清丙氨酸氨基转移酶(ALT)水平；病理组织学结果表明，BJ-JN三个剂量组均使D-GalN诱导的肝细胞损伤减轻，但无显著性差异。
2．BJ-JN对CCl_4致大鼠肝纤维化的保护作用
BJ-JN三个剂量均能使CCl_4升高的血清ALT、天门冬氨酸氨基转移酶(AST)、一氧化氮(NO)和肝脏丙二醛(MDA)降低，使CCl_4降低的血清白蛋白／球蛋白(A／G)、肝脏超氧化物歧化酶(SOD)含量、胸腺指数增加；BJ-JN三个剂量均明显减轻CCl_4所致的肝纤维化程度，不仅降低血清透明质酸(HA)含量，而且减轻肝脏纤维化的病理组织学改变，第19周尤为显著；BJ-JN高剂量组可明显抑制肝纤维化大鼠HSC的增殖和胶原分泌。表明本品对大鼠CCl_4性肝纤维化有明显的防治作用。并提示该药抗肝纤维化作用可能与其抗氧化作用、免疫调节作用和抑制HSC的功能有关。
3．对BCG+LPS诱导小鼠免疫性肝损伤的保护作用
BJ-JN三个剂量均可明显降低升高的血清ALT水平，同时改善异常的免疫学指标，使低下的刀豆蛋白A(Concanavalin A，ConA)诱导脾淋巴细胞的增殖反应上升，使过高的腹腔巨噬细胞(MΦ))产生白细胞介素-1(Interleukin-1，IL-1)
安徽医科大学硕士学位论文
和肿瘤坏死因子(Tumor neerosis factor， TNF)水平下降。BJ一州高剂量可明显减
轻肝细胞损伤和间质炎细胞浸润，低剂量与高剂量组均明显促进肝细胞再生。
提示本品对小鼠免疫性肝损伤，有保护作用，该作用可能与其抗炎和免疫调节
作用有关。
4.BJ一JN血清药理学研究
BJ一JN的含药血清对大鼠肝星状细胞株(HSC一T6)的体外研究结果表明，
BJ一JN(1 2 9 .kg一，)的含药血清可以显著抑制Hsc一T6细胞的增殖和胶原分泌，提
示该品的含药血清对HSC一T6细胞有直接抑制作用。
体内外实验结果综合表明，对D~Ga1N、CC14与BCG十LPS分别诱导小鼠急
性肝损伤、大鼠肝纤维化与小鼠免疫性肝损伤，BJ一JN不仅具有明显的防治作用，
而且有抗氧化和纠正异常的免疫功能的作用，还可明显抑制肝纤维化大鼠HSC的
增殖和胶原分泌。BJ一州含药血清体外可直接抑制HSC一T6细胞的增殖和胶原分
泌。提示本方的抗肝损伤作用与其抗氧化、抗炎、和免疫调节等作用有关。HSC
可能是BJ一州产生抗肝纤维化的的作用靶点之一。
To explore the effect of BJ-JN on liver injury and liver fibrosis, several experimental liver injury models were adopted. The protective mechanism of BJ-JN (3, 6, 12 g. kg-1 d-1) on hepatic damage was analyzed preliminarily. The method of serum pharmacology was used to observe the effect of drug-contained serum of BJ-JN on HSC-T6 cell line in vitro. The main results are as follows.
1. Effect of BJ-JN on D-GalN induced acute liver injury in mice
The liver index and the serum content of ALT induced by D-GalN were significantly reduced by high dose of BJ-JN. Three dosage of BJ-JN alleviated the degree of hepatocytes injury induced by D-GalN in pathological examination.
2. Effect of BJ-JN in CCl4 induced chronic liver injury in rats
The elevation of serum ALT, AST, MDA and NO level were significantly reduced and the reduction of liver T-SOD content, serum A/G and thymus index was increased by BJ-JN. Three dosage of BJ-JN obviously alleviated the degree of liver fibrosis induced by CCl4. Not only the elevation of HA level in serum was reduced, but also the severity of liver fibrosis was significantly relieved in pathological examination, especially in rats injured by CCl4 for 19 weeks. The proliferation and collagen secreting of HSC isolated from liver fibrotic rats were remarkably inhibited by high-dose BJ-JN treatment. The results showed that BJ-JN has obviously anti-fibrosis effect and the mechanism of action might be related to its anti-oxidative activity, immune regulation and the inhibitory effect on HSC function.
3. Effect of BJ-JN on BCG+LPS induced immunologyical liver injury in mice
The elevation of serum ALT content was obviously reduced by three dosage of BJ-JN. It was also found that BJ-JN could restore the diminished splenocyte proliferation induced by ConA and reduced IL-1 and TNF-a production by peritoneal macrophages(M ) in BCG + LPS injured mice. The high-dose BJ-JN obviously alleviated the degree of hepatocytes injury and the infiltration of inflammatory cell induced by BCG+LPS. BJ-JN remarkably promoted hepatocytes regeneration in pathological examination. These results showed that BJ-JN not only has the protective effective, but also has anti-inflammatory and immunoregulatory effects on immunologyical liver injury in mice.
4. The serum pharmacology study of BJ-JN
The results showed that the drug-contained serum of BJ-JN (12 g. kg-1) significantly
restrained the proliferation and collagen secreting of HSC-T6 cell in vitro.
To sum up, BJ-JN has the protective effects on three experimental models (acute, chronic and immunological liver injury). The results in vivo and in vitro suggested that its mechanism of action might be associated with the anti-oxidative activity, immunoregulation and anti-inflammatory effect as well as the inhibitory effect on the function of HSC.
1．BJ-JN对D-GalN诱导小鼠急性肝损伤的保护作用
BJ-JN高剂量组可明显降低D-GalN升高的小鼠肝脏指数和血清丙氨酸氨基转移酶(ALT)水平；病理组织学结果表明，BJ-JN三个剂量组均使D-GalN诱导的肝细胞损伤减轻，但无显著性差异。
2．BJ-JN对CCl_4致大鼠肝纤维化的保护作用
BJ-JN三个剂量均能使CCl_4升高的血清ALT、天门冬氨酸氨基转移酶(AST)、一氧化氮(NO)和肝脏丙二醛(MDA)降低，使CCl_4降低的血清白蛋白／球蛋白(A／G)、肝脏超氧化物歧化酶(SOD)含量、胸腺指数增加；BJ-JN三个剂量均明显减轻CCl_4所致的肝纤维化程度，不仅降低血清透明质酸(HA)含量，而且减轻肝脏纤维化的病理组织学改变，第19周尤为显著；BJ-JN高剂量组可明显抑制肝纤维化大鼠HSC的增殖和胶原分泌。表明本品对大鼠CCl_4性肝纤维化有明显的防治作用。并提示该药抗肝纤维化作用可能与其抗氧化作用、免疫调节作用和抑制HSC的功能有关。
3．对BCG+LPS诱导小鼠免疫性肝损伤的保护作用
BJ-JN三个剂量均可明显降低升高的血清ALT水平，同时改善异常的免疫学指标，使低下的刀豆蛋白A(Concanavalin A，ConA)诱导脾淋巴细胞的增殖反应上升，使过高的腹腔巨噬细胞(MΦ))产生白细胞介素-1(Interleukin-1，IL-1)
安徽医科大学硕士学位论文
和肿瘤坏死因子(Tumor neerosis factor， TNF)水平下降。BJ一州高剂量可明显减
轻肝细胞损伤和间质炎细胞浸润，低剂量与高剂量组均明显促进肝细胞再生。
提示本品对小鼠免疫性肝损伤，有保护作用，该作用可能与其抗炎和免疫调节
作用有关。
4.BJ一JN血清药理学研究
BJ一JN的含药血清对大鼠肝星状细胞株(HSC一T6)的体外研究结果表明，
BJ一JN(1 2 9 .kg一，)的含药血清可以显著抑制Hsc一T6细胞的增殖和胶原分泌，提
示该品的含药血清对HSC一T6细胞有直接抑制作用。
体内外实验结果综合表明，对D~Ga1N、CC14与BCG十LPS分别诱导小鼠急
性肝损伤、大鼠肝纤维化与小鼠免疫性肝损伤，BJ一JN不仅具有明显的防治作用，
而且有抗氧化和纠正异常的免疫功能的作用，还可明显抑制肝纤维化大鼠HSC的
增殖和胶原分泌。BJ一州含药血清体外可直接抑制HSC一T6细胞的增殖和胶原分
泌。提示本方的抗肝损伤作用与其抗氧化、抗炎、和免疫调节等作用有关。HSC
可能是BJ一州产生抗肝纤维化的的作用靶点之一。
To explore the effect of BJ-JN on liver injury and liver fibrosis, several experimental liver injury models were adopted. The protective mechanism of BJ-JN (3, 6, 12 g. kg-1 d-1) on hepatic damage was analyzed preliminarily. The method of serum pharmacology was used to observe the effect of drug-contained serum of BJ-JN on HSC-T6 cell line in vitro. The main results are as follows.
1. Effect of BJ-JN on D-GalN induced acute liver injury in mice
The liver index and the serum content of ALT induced by D-GalN were significantly reduced by high dose of BJ-JN. Three dosage of BJ-JN alleviated the degree of hepatocytes injury induced by D-GalN in pathological examination.
2. Effect of BJ-JN in CCl4 induced chronic liver injury in rats
The elevation of serum ALT, AST, MDA and NO level were significantly reduced and the reduction of liver T-SOD content, serum A/G and thymus index was increased by BJ-JN. Three dosage of BJ-JN obviously alleviated the degree of liver fibrosis induced by CCl4. Not only the elevation of HA level in serum was reduced, but also the severity of liver fibrosis was significantly relieved in pathological examination, especially in rats injured by CCl4 for 19 weeks. The proliferation and collagen secreting of HSC isolated from liver fibrotic rats were remarkably inhibited by high-dose BJ-JN treatment. The results showed that BJ-JN has obviously anti-fibrosis effect and the mechanism of action might be related to its anti-oxidative activity, immune regulation and the inhibitory effect on HSC function.
3. Effect of BJ-JN on BCG+LPS induced immunologyical liver injury in mice
The elevation of serum ALT content was obviously reduced by three dosage of BJ-JN. It was also found that BJ-JN could restore the diminished splenocyte proliferation induced by ConA and reduced IL-1 and TNF-a production by peritoneal macrophages(M ) in BCG + LPS injured mice. The high-dose BJ-JN obviously alleviated the degree of hepatocytes injury and the infiltration of inflammatory cell induced by BCG+LPS. BJ-JN remarkably promoted hepatocytes regeneration in pathological examination. These results showed that BJ-JN not only has the protective effective, but also has anti-inflammatory and immunoregulatory effects on immunologyical liver injury in mice.
4. The serum pharmacology study of BJ-JN
The results showed that the drug-contained serum of BJ-JN (12 g. kg-1) significantly
restrained the proliferation and collagen secreting of HSC-T6 cell in vitro.
To sum up, BJ-JN has the protective effects on three experimental models (acute, chronic and immunological liver injury). The results in vivo and in vitro suggested that its mechanism of action might be associated with the anti-oxidative activity, immunoregulation and anti-inflammatory effect as well as the inhibitory effect on the function of HSC.
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