| Objective:Rouganxiaozheng drink which is composed of nutgrass galingale rhizome, green tangerine peel,fresh membranous milkvetch root, parched largehead atractylodes rhizome, white peony root, common burreed rhizome, zedoary, eupolyphaga seu steleophaga,aromatic turmeric root-tuber, turmeric root tuber,dan-shen root, et al. possesses the function of eliminating blood stasis and lumps, relieving the depressed liver, soothing the liver, regulating the circulation of Qi, and repleniching Qi to invigovate the spleen,which was the effective prescription screened by professor through many years clinical practice and has the notable curative effect on treating hepatic cirrhosis.Further for probing the functional mechanism of Rouganxiaozheng drink, reproduced the rats models of cirrhosis adopting complex factors,then supplied with drug treatment,to study the functional mechanism of protecting hepatic function through observing the effect of Rouganxiaozheng drink on aspartate aminotransferase(AST),alanine aminotransferase (ALT), Total protein(TP),and albumin(Alb); to study the functional mechanism of resisting hepatic fibrogenesis through observing the effect of Rouganxiaozheng drink on Procollagen typeⅢ(PCⅢ), TypeⅣcollagen(Ⅳ-C), Hyaluronic (HA), Laminin (LN)and basic fibroblast growth facter(bFGF); to observe the effect of Rouganxiaozheng drink on pathomor -phology of live tissue through HE and Masson staining.Methods:60 SD rats,half male and half female,among which 48 rats were randomly selected for making model group and the rest were as normal group after normal feeding for 1 week。At first time, the making model group were subcutaneous injected by solution of 40% CCl4-arachisoil, 0.5 ml per 100 gram, afterward, 0.3 ml per 100 gram, two times each week, total 9 weeks. The rats were fed by mix animal feeds of 20% lard, 0.5% cholesterol, and 79.5% maize flour for first 2 weeks, then were fed by mix animal feeds, 0.5% cholesterol, and 99.5% maize flour for the later 7 weeks. Ethyl alcohol was as drinks for rats drinking during whole period of making model. The mormal group was fed by common animal feeds and clean water. On the end of 9th week, success of making model was affirmed.40 Wistar cirrhotic rats were randomly divided into 4 groups, 10 rats per group, female and male were half.①Normal group: watering- stomach with saline.②Model group: watering-stomach with saline.③High-dose Rouganxiaozheng drink group: 2.9 gram per 100 gram, watering-stomach with Rouganxiaozheng drink suspension.④Low-dose Rouganxiaozheng drink group:1.45 gram per 100 gram, watering-stomach with Rouganxiaozheng drink suspension.⑤control group: 0.1 gram per 100 gram, watering-stomach with suspension of complex Biejiaruangan tablet. From the second day after the end of making models, all rats were continuous watering with saline or medicine for 4 weeks one time each day, and were anesthetized with 10% chloral hydrate(0.2ml/kg)after the last time medication and fasted for 12 hr.Afterward,incised in abdominal middle, obtained blood from abdominal aorta,blood centrifuged with hypothermia,and serum separated,then to detemined the contents of ALT, AST, TP,Alb in serum;detemined the contents of PCⅢ,Ⅳ-C,LN,HA in serum by method of radiation-immunization; and detemined the changing of bFGF in hepatic tissue by method of euzymelinked immunosorbent assay. Meanwhile,took out the animal liver,taken hepatic tissue about 1cm×1cm×1cm size from the largest foliage of liver,which were fixed with 4% citromint,embedded with paraffin commonly,and maken slice,to observe the changes of pathomorphology of live tissue through HE and Masson staining.Results1 The effect of Rouganxiaozheng drink on contents of ALT, AST, TP and Alb in serum of cirrhosis rats The ALT and AST contents of serum in model group increased obviously than that of normal group(P<0.01),but the contents of TP and Alb in model group lowered notablely than that of normal group(P < 0.01).After medication,in each treatment groups the ALT and AST contents of serum were lower and the TP and Alb contents of serum higher,high-dose group and low-dose group than that of model group had statistics meaning(P<0.01).The contents of AST and Alb in low-dose group than that of model group had statistics meaning(P<0.01).The contents of Alb in low-dose group than that of control group had statistics meaning (P<0.01). (Table 2)2 The effect of Rouganxiaozheng drink on pathomor -phology changes of liver tissue and proliferation degree of collagenous fibers of liver in cirrhosis rats①The effect of Rouganxiaozheng granule on pathomor -phology changes of liver tissue in cirrhosis rats:In normal group, the liver of rats was soft and rosy, the capsule was smooth and glossy. In model group, hepatic volume had augmented, liver quality had changed hard, the color was gloomy red, lesser tubercle were evident in section. In treatment groups, the volume, quality color and luster had obviously improved, which approached to normal group, and had obvious difference compared to model group.②In normal group, the structure of liver tissue was integrated and darity,the structure of hepatic lobufe was common,and hepatocyte ranked regulation as strip shape and its distribution took on radiation surrounding the central vein.The structure of hepatic sinusoid was darity and has no abnormal changes. Hepatocyte showed polygon, he starch of cells was well distributed, and the shape of nucleus was normal. In model group, the normal structure of hepatic lobufe disappeared, the extensive fibrous tissue of liver interstitial was hyperplasia, which divided the normal hepatic lobufe into inequality group of hepatocyte (fake lobulation), and the diffusive lymphocyte infiltrated in interstitial. The volume of hepatocyte enlarged obviously, partly hepatocyte appeared fatty degeneration critically, and it is clearly to see the regenerated hepatocyte (a couple nuclear). In each treatment group, the degree of hepatic injury was lighter than that of model group. In high-dose group, the normal structure of hepatic lobufe existed, structure of minority hepatic lobufe damaged yet, minority hepatocyte degenerated, cellular necrosis viewed by chance, less lymphocyte infiltrated, a small quantity of fibrous tissue proliferated, no fibrous intermission formed. In low-dose group, structure of hepatic lobufe damaged, part hepatocyte degenerated, accompanying cellular necrosis, more lymphocyte and monocyte infiltrated, fibrous tissue proliferated, but less fibrous intermission formed. The pathomorphology changes of control group were similar to low-dose group.③The effect of Rouganxiaozheng granule on proliferation degree of collagenous fibers in cirrhosis rats:Proliferation degree of collagenous fibers obviously heightened in model group,which had notable difference(P<0.01)compared to that of normal group.After medication,proliferation degree of collagenous fibers in each treatment group were lower than that of model group(P<0.05),above all,the affect of high-dose group was most excellent,whose proliferation degree of collagenous fibers were lower than that of low-dose group and control group(P<0.05). Rouganxiaozheng drink could decrease the proliferation degree of collagenous fibers,whose effect was super to that of complex Biejiaruangan tablet. (Table 1) 3 The effect of Rouganxiaozheng drink on PCⅢ,Ⅳ-C,HA and LN contents of serum in cirrhosis ratsThe contents of serum PCⅢ,Ⅳ-C,HA and LN in model group were higher than that of normal group(P<0.05).After medication,the contents of serum PCⅢ,Ⅳ-C,HA and LN in high-dose group and control group were lower than that of model group(P < 0.05),and the contents of serum PCⅢ,Ⅳ-C in low-dose group were lower than that of model group(P<0.05), but there was no obvious difference on HA and LN(P>0.05). The contents of serumⅣ-C in high-dose group decreased than that of control group.(Table 3)4 The effect of Rouganxiaozheng drink on bFGF of liver tissue in cirrhosis ratsThe contents of bFGF in model group were higher than that of normal group(P<0.01). After medication,the contents of bFGF in each treatment group were lower than that of model group(P < 0.01 or P < 0.05). There was obvious statistical significance of bFGF content between high-dose group and control group(P<0.01). From this we can see that the effert on bFGF of Rouganxiaozheng drink was better than complex Biejiaruangan tablet.(Table 4)ConclusionRouganxiaozheng drink which is composed of nutgrass galingale rhizome, green tangerine peel, fresh membranous milkvetch root, parched largehead atractylodes rhizome, white peony root, common burreed rhizome, zedoary, eupolyphaga seu steleophaga, aromatic turmeric root-tuber, turmeric root tuber, dan-shen root, et al. possesses the function of eliminating blood stasis and lumps, relieving the depressed liver, soothing the liver, regulating the circulation of Qi,and repleniching Qi to invigovate the spleen.1 Rouganxiaozheng drink can obviously lower ALT and AST contents, raise TP and Alb contents of serum, thereby improve hepatic function, and protect liver.2 Rouganxiaozheng drink possesses the function of decreasing necrosis degree of hepatocyte, protecting hepatocyte, promoting the regenerate of hepatocyte, and restraining the synthesis of collagen.3 Rouganxiaozheng drink can obviously decrease the level of PCⅢ,Ⅳ-C,HA,LN in serum .Thereby, Rouganxiaozheng drink poessess the function of hindering and inversing hepatic fibrosis and pathological processes of cirrhosis.4 Rouganxiaozheng drink can lower the content of bFGF in liver tissue of cirrhosis rats, inhibit the hyperplasy of fiber, and thus achieve the function of treating cirrhosis. |
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