Traditional Chinese medicine composition for treating scleroderma

Abstract

The invention provides a traditional Chinese medicine composition for treating scleroderma. The traditional Chinese medicine composition is prepared from, by weight, 25-35 parts of radix rehmanniae preparata, 15-20 parts of honeysuckle flowers, 12-20 parts of honeysuckle stems, 9-15 parts of earthworms, 12-20 parts of radix scrophulariae, 3-6 parts of deer-horn glue, 3-6 parts of baked ginger, 3-6 parts of cassia twigs, 3-6 parts of herba ephedrae, 6-9 parts of radix angelicae sinensis and 3-6 parts of honey-fried licorice roots. The traditional Chinese medicine composition is free of toxic and side effects, quick in treatment effect, high in cure rate and capable of improving skin fibrosis, improving skin pathological change and reducing skin corium layer thickness, and has a good curative effect on systemic scleroderma and localized scleroderma.

Description

Technical field

[0001] The present invention relates to the technical field of traditional Chinese medicine compounds, and in particular to a traditional Chinese medicine composition for treating systemic and localized scleroderma. [Background Technology]

[0002] Systemic sclerosis (SSc) is a chronic, progressive autoimmune disease characterized by inflammatory fibrosis and small vessel lesions affecting the skin or multiple systems. The clinical manifestations of this disease vary in severity, with lesions limited to the skin or subcutaneous tissue in mild cases and multiple systems involved in severe cases, resulting in a poor prognosis. The main manifestations are diffuse hardening and thickening of the skin, or even atrophy, Raynaud's phenomenon, hyperpigmentation, vascular damage and activation, fibrosis of the skin and internal organs, and immune system dysfunction, ultimately leading to death from lung infection and heart and kidney failure. Its pathological process is complex, and its pathogenesis remains unclear. Western medicine currently lacks effective drugs, other than hormones, to cure or alleviate the disease's progression.

[0003] Studies have found that abnormal activation of the Wnt / β-catenin signaling pathway is closely associated with fibrotic lesions in the skin and visceral organs such as the heart, liver, and lungs. Therefore, the Wnt / β-catenin signaling pathway may play a key role in the pathogenesis of SSc. Fibrosis is caused by the excessive synthesis and deposition of extracellular matrix (ECM) proteins such as collagen. The pathogenesis of scleroderma fibrosis is multifactorial, involving multiple cellular pathways that contribute to the development and progression of fibrosis. Recent evidence suggests that Wnt proteins have a potentially important role in fibrosis, and that β-catenin, as a key hub in the pro-fibrotic Wnt signaling pathway, plays a crucial role in the pathogenesis and progression of scleroderma.

[0004] The transduction of the Wnt3a / β-Catenin-GSK-3β / c-myc signaling pathway depends on the level of intracellular β-Catenin. The key lies in whether there is a large amount of structurally stable and soluble β-Catenin in the cytoplasm. The translocation of the hub molecule β-Catenin from the cytoplasm to the nucleus is a sign that the signaling pathway is activated and functions.

[0005] In the absence of Wnt protein signaling, the so-called "β-catenin degradation complex," namely the Axin-APC-CKIα-GSK-3β complex, phosphorylates β-catenin, promoting its subsequent degradation. However, when Wnt protein signaling is enhanced, it binds to its receptor, disrupting the β-catenin degradation complex, thereby reducing β-catenin phosphorylation and ubiquitination degradation. Non-phosphorylated β-catenin accumulates in the cytoplasm and translocates into the nucleus, where it binds to TCF / LEF to form a transcriptional complex, activating the transcription of downstream target genes such as c-myc and Cyclin D1, leading to cell proliferation and differentiation. c-Myc is a nuclear transcription factor. c-Myc mRNA is an important downstream target gene of β-Catenin. The encoded product is mainly involved in the regulation of cell division, differentiation and growth cycle. Under normal conditions, cells do not produce or only express a small amount of c-Myc. Low levels of c-Myc can inhibit cell proliferation and induce cell differentiation. When the expression of the c-Myc gene is abnormally increased, it can drive fibroblasts away from normal growth and proliferate with high proliferative potential.

[0006] Traditional Chinese Medicine doesn't have a specific name for systemic sclerosis. Based on symptoms such as hardened and thickened skin, stiff and swollen fingers, and pain in the limbs and joints, it is classified as "bi syndrome." The cause of this disease is closely related to constitution, lifestyle, climate, and dietary habits. The root cause of the disease lies in the obstruction of the meridians by pathogenic qi.

[0007] Based on years of clinical practice, we believe that insufficient Qi and blood, spleen and kidney Yang deficiency, inability to warm the body, and a weakened defense system are the inherent foundations of systemic scleroderma. This is conditioned by external pathogenic factors such as wind, cold, and dampness, which generate internal heat, phlegm, and blood stasis. This is also related to emotional stimulation and dietary irregularities. Innate deficiency and acquired imbalance allow pathogenic Qi to invade, lingering in the tendons, joints, and muscles of the limbs, resulting in a stagnation of defense and obstruction of the meridians. While the disease is characterized by cold, dampness, and blood stasis as external conditions, its internal foundation lies in impaired organ function and insufficient vital Qi. Wind, cold, dampness, and blood stasis obstruct the body externally, causing illness. Prolonged illness leads to a decline in kidney Yang, allowing internal Yin and cold to accumulate throughout the body's skin, causing a toughened skin and even life-threatening conditions.

[0008] Therefore, based on previous studies, this experiment used subcutaneous injection of bleomycin (BLM) to prepare SSc model mice, and further observed the effects of the traditional Chinese medicine composition on the gene and protein expression of Wnt3a, GSK-3β, β-Catenin, and c-myc in the skin of scleroderma mice from the perspective of the Wnt / β-Catenin pathway, providing a basis for the clinical treatment of scleroderma skin lesions and fibrosis with the traditional Chinese medicine composition of the present invention. [Summary of the invention]

[0009] The technical problem to be solved by the present invention is to provide a traditional Chinese medicine composition for treating scleroderma, which has no toxic side effects, has a rapid therapeutic effect, a high cure rate, can improve skin fibrosis, improve skin pathological changes, reduce the thickness of the skin dermis, and has a good therapeutic effect on both systemic scleroderma and localized scleroderma.

[0010] The present invention is achieved in that:

[0011] A traditional Chinese medicine composition for treating scleroderma comprises the following raw materials in parts by weight: 25-35 parts of cooked rehmannia, 15-20 parts of honeysuckle, 12-20 parts of honeysuckle vine, 9-15 parts of earthworms, 12-20 parts of Scrophularia, 3-6 parts of antler glue, 3-6 parts of roasted ginger, 3-6 parts of cassia twig, 3-6 parts of ephedra, 6-9 parts of angelica, and 3-6 parts of roasted liquorice.

[0012] Furthermore, the traditional Chinese medicine composition includes the following raw materials in parts by weight: 30 parts of cooked rehmannia, 18 parts of honeysuckle, 16 parts of honeysuckle vine, 12 parts of earthworms, 15 parts of Scrophularia, 5 parts of antler glue, 5 parts of roasted ginger, 5 parts of cinnamon twig, 5 parts of ephedra, 7 parts of angelica and 5 parts of roasted licorice.

[0013] The present invention has the following advantages:

[0014] The prescription of the present invention uses Rehmannia root to warm the blood, replenish the essence and benefit the marrow, and is matched with antler glue which warms the kidney yang, benefits the essence and blood, and strengthens the tendons and bones, forming a double tonification of yin and yang, which are the main medicines; assisted by roasted ginger to warm the middle and dispel cold, help yang and dredge the meridians; cinnamon twig to warm and dredge the meridians, help yang and transform qi; angelica to replenish blood and invigorate blood circulation, combined with honeysuckle vine and earthworm to clear away heat and detoxify, harmonize blood and dredge the meridians, and lead Rehmannia root and antler glue directly into the ground, which are the assistant medicines; honeysuckle and Scrophularia ningpoensis to clear away heat and detoxify, nourish Yin-reducing fire can both assist Rehmannia glutinosa in nourishing yin and counterbalance the warming and drying effects of cinnamon twig and roasted ginger. Combined with angelica and roasted licorice root, it forms Simiao Yong'an Decoction, a decoction that studies have shown to significantly reduce vascular inflammation. Ephedra, with its pungent and warm properties, can reach the skin and membranes, unblocking the meridians. Compatible with warming herbs, it opens the pores, dissipates cold accumulation, and draws yang energy from the interior to the exterior, achieving a dispersing effect, serving as an adjuvant. Roasted licorice root harmonizes the various herbs, tonifies the spleen and replenishes qi, and mitigates the pungent and dispersing effects of ephedra and cinnamon twig, serving as a guiding herb. The entire formula nourishes yin and blood while warming yang and strengthening the spleen, resolving phlegm and unblocking the meridians. It tonifies without stagnating, warms and disperses without damaging vital energy, and collectively achieves the functions of tonifying the kidneys and replenishing essence, warming yang and dispersing cold, and clearing heat and unblocking the meridians. Combining the principles that "meridians govern qi, while collaterals govern blood," and that "initially qi stagnates in the meridians, but over time, blood damages the collaterals," it has demonstrated excellent efficacy in treating both systemic and localized scleroderma.

[0015] Animal test results show that the Chinese herbal composition of the present invention has a good therapeutic effect on scleroderma model mice, with rapid onset of action, showing significant effects after 4 weeks of treatment. After 4 weeks of treatment, the weight of the mice increased slightly compared to before treatment, and the negative rate of antinuclear antibodies (ANA) in the mouse serum reached 75%. The thickness of the epithelial skin of the mice was significantly more uniform, the dermis was slightly thinner, the collagen fiber bundles were loosely arranged, the subcutaneous fat layer was slightly thinner, the degree of collagen fiber proliferation in the dermis was reduced, the collagen fiber bundles were loosely arranged, inflammatory cell infiltration was slightly alleviated, and the fat layer was thicker than the model group. Therefore, the present invention can improve skin fibrosis, improve skin pathological changes, and reduce the thickness of the dermis. The hydroxyproline content of mouse skin tissue was significantly reduced, the expression level of Wnt3a in mouse skin was unchanged, and the mRNA and protein expression of β-Catenin and c-myc were upregulated. Clinical efficacy results show that the Chinese herbal composition of the present invention can significantly improve the therapeutic effect of systemic scleroderma.

Brief Description of the Drawings

[0016] The present invention will be further described below with reference to the accompanying drawings and embodiments.

[0017] Figure 1 These are HE staining (×40) images of skin tissues of various groups in the examples of the present invention, wherein a: normal group; b: control group; c: model group; d: Chinese medicine group; e: dexamethasone group.

[0018] Figure 2 These are Masson staining (×4) images of skin tissues of various groups in the examples of the present invention, wherein a: normal group; b: control group; c: model group; d: Chinese medicine group; e: dexamethasone group.

[0019] Figure 3 Schematic diagram of the mRNA and protein expression levels of Wnt3a, GSK-3β, β-Catenin, and c-myc in skin tissues of each group in the examples of the present invention.

[0020] Figure 4 The electrophoresis diagrams are of the protein expression levels of Wnt3a, GSK-3β, β-Catenin, and c-myc in the skin tissues of each group in the examples of the present invention, wherein the lanes from left to right are the normal group, control group, model group, traditional Chinese medicine group, and dexamethasone group, respectively. [Specific implementation method]

[0021] The present invention relates to a traditional Chinese medicine composition for treating scleroderma. The traditional Chinese medicine composition comprises the following raw materials in parts by weight: 25-35 parts of cooked rehmannia, 15-20 parts of honeysuckle, 12-20 parts of honeysuckle vine, 9-15 parts of earthworms, 12-20 parts of Scrophularia, 3-6 parts of antler glue, 3-6 parts of roasted ginger, 3-6 parts of cassia twig, 3-6 parts of ephedra, 6-9 parts of angelica and 3-6 parts of roasted liquorice.

[0022] Preferably, the traditional Chinese medicine composition comprises the following raw materials in parts by weight: 30 parts of cooked rehmannia, 18 parts of honeysuckle, 16 parts of honeysuckle vine, 12 parts of earthworms, 15 parts of Scrophularia, 5 parts of antler glue, 5 parts of roasted ginger, 5 parts of cinnamon twig, 5 parts of ephedra, 7 parts of angelica and 5 parts of roasted liquorice.

[0023] The following will be combined with the Figure 1-4 The technical solutions of the present invention are clearly and completely described in the following and in detail. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative work are within the scope of protection of the present invention. If specific conditions are not specified in the examples, the experiments were carried out under conventional conditions or the conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments is not specified, they are all conventional products that can be purchased commercially.

[0024] Example 1

[0025] A solution of a traditional Chinese medicine composition for treating scleroderma, the specific preparation process is as follows:

[0026] Prepare 30g of Rehmannia glutinosa, 5g of antler glue, 3g of roasted ginger, 5g of cinnamon twig, 5g of ephedra, 15g of honeysuckle, 15g of Scrophularia ningpoensis, 6g of angelica, 3g of roasted liquorice, 15g of honeysuckle vine, and 9g of earthworm. Decoction in water, filter, concentrate to a fluid extract, extract with ethanol, filter, recover ethanol, and adjust the final filtrate to a crude drug content of 1.12g / ml with water. Dispense and sterilize.

[0027] Example 2 Animal Experiment

[0028] 1 Animals: 40 healthy female BALB / C mice, SPF grade, female, 6 weeks old, weighing 20±2 g.

[0029] 2. Drugs: Traditional Chinese Medicine (the traditional Chinese medicine solution prepared in Example 1). Bleomycin hydrochloride for injection was dissolved in autoclaved physiological saline to a maximum concentration of 500 μg·mL. -1 (Imported drug registration number: H20090885, manufactured by Nippon Kayaku Co., Ltd.) Dexamethasone acetate tablets were dissolved in normal saline to a concentration of 0.2 mg mL -1 Dexamethasone acetate suspension (National Medicine Approval Number: H35020317).

[0030] 3 Methods

[0031] 40 mice with 1*1cm hair in the central area of the back were randomly divided into a normal group, a blank group of 8 mice each, and a modeling group of 24 mice. The normal group did not receive any injection, the blank group received a subcutaneous injection of 0.1mL of normal saline on the back, and the modeling group received a subcutaneous injection of 500μg / mL bleomycin 0.1mL / d for 4 consecutive weeks to establish a scleroderma model. After successful modeling, the mice were randomly divided into a model group, a traditional Chinese medicine group, and a dexamethasone group of 8 mice each; the control group and the model group were given normal saline, and the traditional Chinese medicine group and the dexamethasone group were gavaged with traditional Chinese medicine (0.112g·10g -1 ·d -1 ), dexamethasone hydrochloride (0.02mg·10g -1 ·d -1 ), and 4 weeks later, skin tissues were collected and stained with HE. qPCR and Western blot were used to detect the mRNA and protein expressions of β-Catenin in mouse skin, respectively.

[0032] 4 Results

[0033] 4.1 Observation of the general condition of mice before and after treatment

[0034] Before subcutaneous injection of bleomycin for modeling, the general condition of mice in all groups was good, with no significant differences. They ate and drank normally, had normal bowel movements, were responsive, had shiny fur, and had good skin elasticity. After subcutaneous injection of saline in the blank control group, the skin in the injected area showed no significant adhesion to the subcutaneous tissue, the fur had good gloss and elasticity, and the fur had fully grown out. The general condition did not differ significantly from that of the normal control group. However, after subcutaneous injection of bleomycin hydrochloride in the modeling group, the mice gradually became less energetic, drank less readily, had poorer fur gloss and skin elasticity, and gradually lost hair. Fur in the marked area did not grow back, adhered to the subcutaneous tissue, and increased resistance to needle insertion in the skin. All mice in all groups survived four weeks after subcutaneous injection. After modeling, the body weight of the mice in each group remained unchanged, with no statistically significant differences (P>0.05).

[0035] After the start of oral gavage treatment, the skin thickness and hardness at the injection site of the mice in the TCM group decreased to varying degrees compared to the model group. A small amount of fur grew, and the fur glossiness was significantly improved compared to the model group. There were no significant differences in water and food intake compared to the normal group, blank control group, and dexamethasone group. Comparison of weight changes in mice before and after treatment is shown in Table 1.

[0036] After treatment, the body weight of mice in the model and dexamethasone groups decreased compared with the normal group, and the difference was statistically significant (P<0.05). There was no statistically significant difference in the body weight of mice in the other groups (P>0.05). The body weight of the model control group decreased significantly, with a statistically significant difference before and after treatment (P<0.05). The body weight of the TCM group increased slightly compared with before treatment, but the difference was not statistically significant (P>0.05). The body weight of the dexamethasone group decreased compared with before treatment, with a statistically significant difference (P<0.05).

[0037] Table 1 Changes in mouse body weight before and after treatment

[0038]

[0039] Note: a: Compared with the model group: *P<0.05; b: Compared with the normal group: #P<0.05; c: Comparison before and after treatment: △P<0.05

[0040] As can be seen from Table 1, the Chinese medicinal composition for treating scleroderma of the present invention has no toxic or side effects.

[0041] 4.2 Serum antinuclear antibody (ANA) in each group of mice

[0042] Serum antinuclear antibodies (ANA) were positive in 10 of the 40 samples, including 7 in the model control group, 2 in the traditional Chinese medicine group, and 1 in the dexamethasone group. The positive expression rate of ANA in each group was statistically analyzed using the R×C contingency table chi-square test: 2 =22.667, P=0.0001<0.05, indicating that the positive expression rate of ANA in each group was statistically significant (see the table below).

[0043] Table 2 Comparison of the positive rates of antinuclear antibodies (ANA) in the serum of mice in each group before and after treatment ( n=8)

[0044]

[0045] Table 3 Chi-square comparison of the positive rates of antinuclear antibodies (ANA) in the serum of mice in each group before and after treatment

[0046]

[0047] Note: Chi-square split test *P<0.013 is statistically significant

[0048] As can be seen from the table, the negative rate of antinuclear antibodies ANA in the serum of mice treated with the Chinese medicinal composition of the present invention is as high as 75%.

[0049] 4.3 HE staining results of skin tissue

[0050] The results of HE staining of the skin tissues of mice in each group are shown in Figure 1 As shown, from Figure 1 It can be seen that after treatment, the Figure 1 -a) Mouse skin compared to blank control group (see Figure 1 -b) had no significant difference, the model group (see Figure 1 -c) It can be seen that the thickness of the epithelial layer is significantly different, the dermis is thickened and disordered, and the subcutaneous fat layer is thinned. Figure 1 -d) The thickness of the epithelial layer was relatively uniform, the dermis was slightly thinner, the collagen fiber bundles were loosely arranged, and the subcutaneous fat layer was slightly thinner; in the dexamethasone group (see Figure 1 -e) The proliferation of collagen fibers in the dermis was improved, and the atrophy of hair follicles was alleviated.

[0051] 4.4 Masson staining results of mouse skin tissue in each group

[0052] The results of Masson staining of the skin tissues of mice in each group are shown in Figure 2. Figure 2 As shown, from Figure 2 It can be seen that after treatment, the Figure 2 -a) Mouse skin compared to blank control group (see Figure 2 -b) had no significant difference, the model group (see Figure 2 -c) showed proliferation of collagen fibers in the dermis, disordered arrangement, infiltration of inflammatory cells, atrophy of hair follicles, and thinning of the subcutaneous fat layer; the Chinese medicine group (see Figure 2 -d) It can be seen that the degree of collagen fiber proliferation in the dermis was alleviated to a certain extent, the arrangement of collagen fiber bundles was relatively loose, the infiltration of inflammatory cells was alleviated to a certain extent, the atrophy of hair follicles was improved, and the fat layer was thicker than that of the model group; the dexamethasone group (see Figure 2 -e) The proliferation of collagen fibers in the dermis is improved, the arrangement is loose and disordered, and the atrophy of hair follicles is improved.

[0053] 4.5 Hydroxyproline content in skin tissue of mice in each group

[0054] Table 4 shows the content of hydroxyproline in the skin tissue of mice in each group. It can be seen from Table 4 that compared with the normal group, the hydroxyproline content in the model group and the dexamethasone group increased, and the difference was statistically significant (P < 0.05), and there was no difference between the blank control group and the Chinese medicine group; compared with the model group, the hydroxyproline content in each group decreased, and the difference was statistically significant (P < 0.05); there was no statistically significant difference between the Chinese medicine group and the dexamethasone group (P> 0.05).

[0055] Table 4 Hydroxyproline content in skin tissue of mice in each group

[0056]

[0057]

[0058] Therefore, the Chinese medicine composition of the present invention can improve the degree of skin fibrosis and the content of hydroxyproline in scleroderma model mice.

[0059] 4.6 Wnt3a, GSK-3β, β-Catenin, and c-myc mRNA and protein expression in skin tissue

[0060] The bar graphs of the mRNA expression levels of Wnt3a, GSK-3β, β-Catenin, and c-myc in the skin tissues of mice in each group are shown in the figure. Figure 3 The protein expression electrophoresis diagram is shown in Figure 4, and the mRNA expression level is shown in Table 5.

[0061] Compared with the normal group, there were no differences in the mRNA and protein expression levels of Wnt3a, GSK-3β, β-Catenin, and c-myc in the blank control group (P>0.05); the mRNA and protein expression levels of Wnt3a, β-Catenin, and c-myc in the model group were increased, and the expression level of GSK-3β was downregulated, and the differences were statistically significant (P<0.05); the expression level of Wnt3a in the TCM group was not different (P>0.05), while the mRNA and protein expression levels of β-Catenin and c-myc were upregulated, and the differences were statistically significant (P<0.05); the mRNA and protein expression levels of Wnt3a and c-myc were upregulated, and the expression of GSK-3β was downregulated in the dexamethasone group, and the differences were statistically significant (P<0.05), while the expression level of β-Catenin was not different (P>0.05). Compared with the model group, the mRNA and protein expression levels of Wnt3a, β-Catenin, and c-myc in the TCM group were downregulated, and the differences were statistically significant (P<0.05). Although the expression level of GSK-3β increased, the difference was not statistically significant (P>0.05). (See Table 5 below, a: Compared with the model control group: *P<0.05; b: Compared with the normal group: #P<0.05)

[0062] Table 5 The mRNA expression levels of Wnt3a, GSK-3β, β-Catenin and c-myc in the skin tissues of mice in each group

[0063]

[0064]

[0065] The Wnt / β-Catenin signaling pathway is closely related to TGF-β signaling and is a key pathway in fibrosis. TGF-β can activate the Wnt / β-Catenin signaling pathway in vitro and in vivo. Reports have shown that nuclear accumulation of β-Catenin can be detected in fibroblasts transfected with TGF-β1 in vitro, and downstream TCF / LEF response elements are activated. In vivo, overexpression of the tissue-activated TGF-β receptor type I (TBRI) induces TGF-β signaling, which leads to nuclear accumulation of β-Catenin and increased transcription of target genes in the skin. Treatment with SD-208, a selective small molecule inhibitor of TGF-β receptor type I, significantly improves Wnt signaling activation in experimental fibrosis models. TGF-β regulates Wnt / β-Catenin signaling by modulating its endogenous inhibitors.

[0066] The results of this experiment suggest that the mRNA and protein expression levels of Wnt3a, GSK3β, β-Catenin, and c-myc in the skin of mice subcutaneously injected with normal saline were no different from those in the normal non-injected group. It can be seen that subcutaneous injection of non-drugs has no effect on the wnt / β-Catenin pathway indicators.

[0067] Experimental results showed that the mRNA expression levels of Wnt3a, GSK-3β, β-Catenin, and c-myc in the skin of the model group were increased, indicating that bleomycin can induce fibrosis in the skin of scleroderma mice. Compared with the model group, the mRNA expression of Wnt3a, β-Catenin, and c-myc was reduced in the traditional Chinese medicine group. It is considered that the traditional Chinese medicine composition of the present invention may inhibit the degree of skin fibrosis by inhibiting the wnt / β-Catenin pathway at the genetic level.

[0068] Experimental results showed that in the skin of model mice subcutaneously injected with bleomycin, Wnt3a, β-Catenin, and c-myc protein expression levels were upregulated, while P-β-Catenin expression was downregulated, further confirming that bleomycin can induce fibrosis. Total GSK-3β protein levels in the model group were no different from those in the normal control group. GSK-3β has two forms, active and inactive, and its activity is related to the site and ratio of phosphorylation. Phosphorylation of GSK-3β at Ser9 inhibits its activity, while phosphorylation at 216 enhances its activity. This suggests that bleomycin-induced inhibition of GSK-3β protein activity in the skin of scleroderma mice inhibits β-Catenin phosphorylation and ubiquitination, leading to an increase in free β-Catenin and downstream c-myc expression, exacerbating fibrosis. This is consistent with other studies showing that inhibition of GSK-3β activity can exacerbate skin fibrosis. This suggests that the traditional Chinese medicine composition of the present invention may inhibit the Wnt / β-Catenin pathway at the protein level, thereby reducing the severity of skin fibrosis.

[0069] According to experimental research results, the efficacy of the present Chinese herbal composition in treating scleroderma may be related to inhibiting the expression of the Wnt / β-Catenin pathway. Specifically, the present Chinese herbal composition can inhibit Wnt3a expression, thereby inhibiting β-Catenin expression, and further inhibiting the expression of the downstream c-myc, thereby delaying the development and progression of SSc fibrosis. These experimental results demonstrate that the present Chinese herbal composition has a relatively good therapeutic effect on SSc model mice, which may be one of the mechanisms of action of the present Chinese herbal composition in treating scleroderma.

[0070] Therefore, the therapeutic effect of the Chinese medicine composition of the present invention on scleroderma mice is related to its ability to inhibit the wnt / β-Catenin pathway.

[0071] Example 3

[0072] Typical clinical cases:

[0073] An 18-year-old female patient presented to the hospital on August 26, 2012. Her chief complaint was difficulty moving her limbs and joints for over three years. Over a year ago, she developed swelling and tightness on her upper limbs, making them difficult to pinch. This swelling gradually spread to her limbs and trunk, resulting in limited joint movement. Her hands turned purple when exposed to cold, which resolved spontaneously after a few minutes. Her face also became red. She was diagnosed with systemic sclerosis at the Union Hospital of Fujian Medical University and treated with methylprednisolone 50 mg / day for anti-inflammatory and immunosuppressive therapy and penicillamine 1 g / day for Raynaud's phenomenon. However, her symptoms recurred with limited efficacy. Symptoms included hardening, atrophy, and cooling of the skin on her limbs and trunk, limited limb movement, and an inability to squat. She also experienced itchy and flushed face. Her lips became thinner, with a preference for warmth and aversion to cold. Her appetite was normal, her stools were loose, her tongue was pale and tender, her tongue coating was white, and her pulse was deep and thready. Physical examination revealed facial flushing, but no obvious rash, swelling, or induration. The skin on both forearms and dorsum of the hands is grayish yellow and waxy, swollen and tight, not easily wrinkled, slightly indented when pressed, with decreased skin elasticity and low skin temperature. Raynaud's sign is positive, and flexion and extension of the knuckles are difficult. Laboratory examination: Blood routine: White blood cell count 20.50*10 9 Neutrophil percentage: 68.36%. Myocardial enzymes: aspartate aminotransferase 93 IU / L, hydroxybutyrate dehydrogenase 473 IU / L, creatine kinase 2769 IU / L, CKMB 116.0 IU / L, lactate dehydrogenase 609 IU / L. Liver function: alanine aminotransferase 163 IU / L, glutamyl transpeptidase 135 IU / L. Myoglobin: 550.93 ng / mL. Complete immune panel: immunoglobulin G: 16.3 g / L, complement C3: 30.75 g / L. ANA+ ANA antibody profile: antinuclear antibody ANA2+ nucleolar, anti-PM-Scl antibody ++. Electromyography: Myogenic damage to the peripheral nerves (mild sensory and motor involvement), high likelihood of scleroderma. Quadriceps muscle biopsy pathological diagnosis: Microscopic examination of the muscle tissue revealed varying degrees of atrophy and deformation of the muscle fibers, with positive Congo red staining. However, striations were still visible in the sarcoplasm, fibrous tissue proliferation was observed between the muscle fibers, and lymphangiocellular infiltration was present. Cardiac ultrasound revealed grade I tricuspid regurgitation with mild pulmonary hypertension. Lung CT scan: 1. Mild bilateral axillary and mediastinal lymphadenopathy; 2. Hepatosplenomegaly. Abdominal ultrasound: 1. Polypoid lesions of the gallbladder; 2. Hepatosplenomegaly; 3. Small pelvic effusion. Traditional Chinese Medicine diagnosis: Bi syndrome, characterized by yang deficiency and cold stagnation. The patient was advised to withdraw penicillamine and reduce methylprednisolone dosage to 30 mg / day, with additional Chinese herbal formulas. Prescription: Rehmannia root 24g, Astragalus root 15g, Deer antler glue 6g, Roasted ginger 10g, Ephedra 4g, Cinnamon twig 4g, White mustard seed 6g, Atractylodes macrocephala 9g, Sophora flavescens thorn 10g, Honeysuckle 15g, Honeysuckle vine 15g, Scrophularia ningpoensis 10g, Earthworm 9g, Roasted Licorice root 3g. One dose per day, divided into morning and evening. After 10 doses, the itching on the face will be relieved, the skin on the limbs and trunk will become slightly softer, and the range of motion will be slightly better, allowing one to squat with some effort.

[0074] Second diagnosis: After three months of treatment with the above prescription and supplemented with spleen-strengthening, blood-nourishing and blood-activating methods, the patient only retained methylprednisolone 5mg / d. The movement of the limbs was basically restored, the skin became soft and could be pinched for the most part, the skin temperature was slightly lower, and the condition improved significantly.

[0075] Two years of follow-up revealed no recurrence. The patient's skin on the limbs and trunk became soft and warm, and he was able to move his limbs freely, including squatting. His complexion was radiant and non-itchy. He was given nausea and vomiting medication, and his bowel movements were regular. His tongue was tender and pale red, with a thin white coating and a slightly thready pulse. Laboratory tests (July 21, 2014) revealed normal blood count, liver function, and myoglobin levels. Complete immunoglobulin panel: Immunoglobulin A: 0.32 g / L, Complement C3: 0.62 g / L. Complete biochemistry panel: Creatine kinase 471 IU / L, CKMB 23.0 IU / L, Cholinesterase 4851 IU / L, ANA+. ANA antibody profile: Antinuclear antibody ANA1+ nucleolar, anti-PM-Scl+, and other parameters were normal. Cardiac ultrasound revealed no significant abnormalities in cardiac structure or function. Lung CT scan: Ground-glass opacities in the lower lobes of both lungs near the posterior chest wall, suggesting interstitial changes. Abdominal ultrasound revealed no significant abnormalities. Quadriceps femoris biopsy pathology: no obvious abnormalities were found.

[0076] The patient was followed up for 12 years with no recurrence. She visited the clinic every 1-2 months. Antinuclear antibodies (ANA1) were positive and anti-PM-Scl antibodies were positive, with only a few skin spots. Her blood and urine tests, complete routine biochemistry panel, and cardiopulmonary structure and function were normal.

[0077] In summary, the Chinese medicine composition of the present invention can improve the degree of skin fibrosis in patients with scleroderma to a certain extent. Considering the hot climate in the southern region and the characteristics of excessive heat toxicity or yin deficiency and latent heat, the classic bleomycin (BLM) was used for subcutaneous injection to prepare scleroderma model mice. The experiment also found that this prescription can improve the degree of skin fibrosis in the model mice: the epidermis and dermis are thinner than those in the model group, the vacuoles between the connective tissue are reduced, the fat cells are increased, and the collagen fibers are thinner and arranged more neatly.

[0078] As can be seen, the results of animal experiments show that the Chinese medicine composition of the present invention has a good therapeutic effect on scleroderma model mice. After 4 weeks of treatment, the weight of the mice increased slightly compared with before treatment, and the negative rate of mouse serum antinuclear antibodies (ANA) reached 75%. The thickness of the epithelial skin of the mice was significantly more uniform, the dermis was slightly thinner, the collagen fiber bundles were loosely arranged, the subcutaneous fat layer was slightly thinner, the degree of collagen fiber proliferation in the dermis was reduced, the collagen fiber bundles were loosely arranged, the inflammatory cell infiltration was slightly alleviated, and the fat layer was thicker than the model group. Therefore, the present invention can improve skin fibrosis, improve skin pathological changes, and reduce the thickness of the dermis layer. The hydroxyproline content of mouse skin tissue was significantly reduced, the expression level of Wnt3a in mouse skin was unchanged, and the mRNA and protein expression of β-Catenin and c-myc were upregulated. Clinical efficacy results show that the Chinese medicine composition of the present invention can significantly improve the therapeutic effect of systemic scleroderma.

[0079] Although the specific embodiments of the present invention are described above, those skilled in the art should understand that the specific embodiments described are merely illustrative and are not intended to limit the scope of the present invention. Equivalent modifications and changes made by those skilled in the art in accordance with the spirit of the present invention should be included within the scope of protection of the claims of the present invention.

Claims

1. A Chinese medicine composition for treating scleroderma, characterized in that: The traditional Chinese medicine composition comprises the following raw materials in parts by weight: 25-35 parts of cooked rehmannia, 15-20 parts of honeysuckle, 12-20 parts of honeysuckle vine, 9-15 parts of earthworms, 12-20 parts of Scrophularia, 3-6 parts of antler glue, 3-6 parts of roasted ginger, 3-6 parts of cassia twig, 3-6 parts of ephedra, 6-9 parts of angelica and 3-6 parts of roasted liquorice.

2. A Chinese medicine composition for treating scleroderma according to claim 1, characterized in that: The traditional Chinese medicine composition comprises the following raw materials in parts by weight: 30 parts of cooked rehmannia, 18 parts of honeysuckle, 16 parts of honeysuckle vine, 12 parts of earthworms, 15 parts of Scrophularia, 5 parts of antler glue, 5 parts of roasted ginger, 5 parts of cassia twig, 5 parts of ephedra, 7 parts of angelica and 5 parts of roasted liquorice.

Images