丹参酚性酸B对大鼠传代肝贮脂细胞增殖、形态和合成细胞外基质的影响

EFFECT OF LITHOSPERMATE B ON PROLIFERATION AND SHAPE OF RAT HEPATIC FATSTORING CELLS AND PRODUCTION OF EXTRACELLULAR MATRIX IN VITRO

目的丹参酚性酸Ｂ（简称丹酚酸Ｂ）是丹参抗肝纤维化的有效水溶性成分。用丹酚酸Ｂ镁盐作用于大鼠传代肝贮脂细胞，观察药物对贮脂细胞增殖、形态和合成细胞外基质功能的影响。方法用链霉蛋白酶和胶原酶原位灌注大鼠肝脏，１１％Ｍｅｔｒｉｚａｍｉｄｅ离心分离贮脂细胞，传一代后加入不同浓度的丹酚酸Ｂ培养７２ｈ，在最后２４ｈ加入［３Ｈ］胸腺嘧啶核苷共同温育，检测其细胞内的掺入量；传一代细胞爬片后，加入不同浓度的丹酚酸Ｂ培养７２ｈ，培养片上细胞用ＰＡＰ法作Ⅰ、Ⅲ和Ⅳ型胶原及板层素免疫组化显色。结果除１０－４ｍｏｌ／Ｌ的丹酚酸Ｂ对贮脂细胞有细胞毒性作用外，１０－５～１０－７ｍｏｌ／Ｌ对［３Ｈ］胸腺嘧啶核苷贮脂细胞内的掺入量都有明显的抑制作用，而对细胞的形态无明显影响（Ｐ＜０．０１；Ｐ＜０．０５）；１０－６ｍｏｌ／Ｌ药物能明显减少贮脂细胞Ⅰ、Ⅲ型胶原和板层素阳性细胞数量（Ｐ＜０．０５；Ｐ＜０．０５；Ｐ＜０．００１），揭示贮脂细胞合成细胞外基质的功能受到抑制。结论丹酚酸Ｂ抗肝纤维化的部分机理在于抑制了活化贮脂细胞增殖，并通过抑制贮脂细胞生成细胞外基质而减少了胶原纤维在肝内的沉积及干扰了狄氏间隙基底膜的形成而减轻了肝窦毛细胞血管化。

Lithospennate B is one kind of water soluble component of Radix Salviae Miltiorrhizae. This article is deal with observation on proliferation and shape of hepatic fat-storing cells(FSC)of rat and its function producing extracellular matrix in subcuItured FSC incubated with Magnesium Lithospermate B(MLB). Methods Perfusing livers of normal adult rats with proteinase E and collagenase in situ, FSC were isolated by centrifugation with 11% metrizamide,(1) The subcultured FSC were incubated with 10￣(-4)～10￣(-7)mol/L of MLB for 72 hours,L-[￣3H]-thymidine was added into the solution of incubation in last 24 hours, then to assay the amount of L-[￣3H]-thymidine incorporated into FSC.(2)FSC were subcuItured on cover glasses and were incubated with 10￣(-4)～10￣(-7) mol/L of MLB for 72 hours. Then the immunohistochemical staining was made for showing type Ⅰ,Ⅲ and IV collagen and laminine(LM). Results Except that 10￣(-4) mol/L of MLB had toxicity to FSC, 10￣(-5)～10￣(-7)mol/L of MLB inhibited the incorporation of L-[￣3H]-thyidine into FSC markedly but had no effect on the shape of FSC (p<0.01;P<0.05). 10￣(-6) mol/L of MLB obviously reduced the number of positive FSC for type Ⅰ, Ⅲ collagen and LM(P<0.05; P<0.05;P<0.001). It was indicated that the function of FSC was inhibited in production of extracellular matrix, Conclusion A part of the effective mechanism of MLB on hepatic fibrosis is what can inhibit the proliferation of active F SC, and FS C to produce extracellular matrix, the deposited collagen fiber in liver was decreased, the degree of capillarization ofsinusoids was reduced.