| Based on the meridian-collateral theory of traditional Chinese medicine, there are many derivative acupuncture& moxibustion (AM) methods. Each method has its own indications and advantages of diseases. When it comes to hepatic fibrosis (HF) ---a chronic pathological process, many AM methods show good effects. However, which one is the most effective? What are the differences among different AM methods? Therefore, in order to answer those questions, the ancient and modern, clinical and experimental AM literatures of HF treatments were summarized and analyzed in this paper. In addition, two frequently-used AM methods--- medicinal vesiculation (MV) and electro-acupuncture (EA) were chosen to stimulate Ganshu acupoint (BL18), a common chosen acupoints, in CCl4 model HF rats, and their effectiveness were compared.The differences between MV and EA include the simulating methods and acting levels in acupoint areas (AA). MV applies chemical irritation on shallow skin layer, while EA simulates acupoints with electricity and acts on deep connective tissues and muscle layer. Researches have indicated that the different tissue structures of acupoints decide and impact the specific effects on acupoints. Mast cells (MCs) is considered to be an important part of the meridian-acupoint dimensional architecture and closely related to meridian functional activities. In meridian and AA, MCs are distributed in specific concentration. MCs are also hierarchically arranged in AA skin layer. Besides, it has been confirmed that different AM can recruit MCs to AA and activate MCs to degranulation. Thus, the different effects of MV and EA on MCs activation and skin morphology, thickness in BL18 were observed upon the different simulations and gradations. The different acting mechanisms were also discussed.The experimental researchObjectiveTo compare the different efficacies of MV and EA on BL18 in HF model rats, and and discuss the different effects of MCs activation in BL18 AA and skin morphology, thickness.Methods40 male SD rats were randomized into normal, model, EA and MV groups (10rats/group). The HF model was made by intraperitoneal injection with CCl4 (40% CCl4 mixed with olive oil,0.3ml/100g per rat, twice per week for 6 weeks). From the third week, the normal group and model group received constraint for 20min, every other day for 4weeks. EA group received EA on BL18 (1mA,50Hz,20min, every other day for 4weeks), and MV group received BL18 point-application with cantharides (0.05g cantharides mixed water into paste, 6hours, twice per week for 4weeks). On the next day after finishing all treatments, the blood was gathered and separated of serum to test PC â…¢, â…£-C, HA and LN. The left lobe of liver was sampled, stained with hematoxylineosin (HE) for the liver pathological observation. The left-hand skin tissues of BL18 were sampled, stained with toluidine blue (TB) for the observation of MCs activation in different layers and AA skin morphology. The skin thicknesses were also measured.Results1 PC â…¢, â…£-C, HA and LN in Serum:PC-â…¢, â…£-C, HA and LN in the model group, were all significantly higher than those of the normal group (P<0.01). There was no obvious difference between the model group and EA group, but PC-â…¢, â…£-C in the later one showed declining trends. In MV group, IV-C and HA were markedly declined (P<0.05). Compared with the EA group, LN in MV group was significantly lower (P<0.05).2 Hepatic Pathology:â‘ Visual observation:Livers in the normal group were fresh red, soft, smooth and having sharp edges. While in the model group, the livers seemed dark red, hard and having granular surface with heavy fat tissues circumvolutio, and the edges were blunt. In the EA group, the livers were dark or deep red, a little bit hard, and showed occasional granular changes with fat tissues adhesion, and the edges were relatively sharp. In the MV group, the livers were deep red, relatively soft, smooth surface with few fat tissues adhered, and having sharp edges.â‘¡ HE staining observation:In the normal group, the livers were with complete hepatic lobule structures, and hepatic cells were tidily and radially arranged around the central veins, with no fibrous connective tissues (FCT) proliferated. In the model group, hepatic lobule structures were damaged and hepatic cells were arranged in disorder with degeneration and necrosis. Obvious FCT hyperplasia was in the portal areas and hepatic lobules and pseudolobule proliferated. Different improvements were observed in the EA and MV groups. In the EA group, hepatic cells were arranged in disorder with some degenerated. FCT hyperplasia was in and around the portal areas and short intervals formed. In the MV group, hepatic cells were regularly arranged, the lobule structures were mostly complete, and little FCT hyperplasia was shown in the portal areas.3 MCs Activities in BL18:â‘ Cutaneous MCs of BL18 total number and degranulated number(rate):a. MCs total number:No obvious difference was shown between model group and normal group; EA group and MV group was significantly increased than model group (P<0.01); no significant difference was shown between EA group and MV group. b. MCs degranulated number:No significant differences were shown between model group and normal group and between EA group and model group; MV group was much higher than model group and EA group (P<0.01,P<0.05). c. MCs degranulated rate:To compared with normal group, model group, EA group and MV group were all significantly raised (P<0.05,P<0.01), and no obvious differences were shown among the three groups.â‘¡ MCs distribution in each cutaneous layer of BL18:a. Comparison within each group:MCs total numbers of subcutaneous layer and dermal reticular layer in each group, MCs degranulated numbers of subcutaneous layer and dermal reticular layer in normal group and model group, MCs degranulated numbers of subcutaneous layer in EA group and dermal reticular layer in MV group were all significantly increased than those in dermal papilla layer(P<0.05 or P<0.01); MCs total numbers of subcutaneous layer in normal group and model group, MCs total and degranulated numbers of subcutaneous layer in EA group were much higher than those in dermal reticular layer(P<0.01), MCs degranulated number of dermal layer in MV group was obvious higher than that in subcutaneous layer(P<0.01). b. Comparison among groups:Between the normal group and model group, the MCs total and degranulated numbers were no obvious differences in each layer; The dermal papilla layer and reticular layer:The MCs total and degranulated numbers of MV group in the two layers were substantially higher than those of model group and EA group(P<0.05 or P<0.01); The subcutaneous layer:MCs total number:EA group and MV group were markedly increased than model group(P<0.01,P<0.05), and no obvious difference between EA group and MV group, MCs degranulated number:Compared with model group, no significant differences were shown in EA group and MV group, while EA group was much higher than normal group and MV group(P<0.01,P<0.05).4 The Skin Morphology and Thickness in BL18:â‘ Skin Morphology:The rats in normal group were with normal skin morphology in AA. The bounds were clear among epidermis multiple layers, and different cells were typical stratified layers arrangement. Dermis collagen fibers arranged in dense and orderliness. Subcutaneous tissues were with moderate fat deposits. In the model group, the epidermis layers were lack of stratified layers arrangement, cells decreased with no uniform sizes, even with vacuoles degeneration. Dermis collagen fibers and subcutaneous fat decreased. In the EA group, the epidermis cell layers diminished, dermis collagen fibers and subcutaneous fat decreased. In the MV group, the cuticle was thin and the epidermis cells were with typical stratified layers arrangement, new vessels and glands appeared in the dermis layer and moderate fat deposits in the subcutaneous layer.â‘¡ Skin Thickness:a. Aggregate Thickness:Compared with normal group, the aggregate thicknesses of the other three groups were all significantly thinner (P<0.01, P<0.01, P<0.05). Compared with model group and EA group, the thickness in MV group was markedly increased (P<0.01). b. Layer Thickness:Compared with normal group, the dermal and subcutaneous layer thicknesses of model group were significantly thinner (P<0.01, P<0.05) and the epidermal layer thickness shown a thinning trend (P=0.07). There was no significant difference in each cutaneous layer thickness between model group and EA group. The each cutaneous layer thickness in MV group was much increased than in model group and EA group (P<0.05 or P<0.01).5 The Correlation Analysis between Skin Thickness in BL18 AA and PC â…¢,â…£-C, HA and LN Indexes:There was a significant negative correlation between the skin total thickness in BL18 AA and â…£-C, HA and LN indexes, and no marked correlation with PCâ…¢.Conclusionsâ‘ Successfully modeled the rats with HF by intraperitoneal injection with CCl4 in the dose of 0.3ml/100g, twice per week for 6 weeks.â‘¡Both MV and EA could improve the serum HF indexes in CCl4 HF model rats in different degrees and reduce FCT hyperplasia and sedimentation in livers, which prompted that both MV and EA have anti-fibrotic effects, and MV showed a better efficacy.â‘¢Both MV and EA could recruit and activate MCs in BL18 AA and MV showed a stronger effect. MV and EA could adjust the distribution of MCs in AA, and MV mad MCs gathered in the dermal papilla layer and dermal reticular layer and EA in the subcutaneous layer.â‘£In CCl4 HF model rats, the skin of BL18 presented pathological changes and significantly thinned, MV could repair the skin hierarchical pathological structures and maintain its integrity, while EA had no effect. The skin thickness was significantly negatively correlated with HF pathological degrees, which meant thinner BL18 skin thickness, heavier HF pathological injury.
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