WO2018209679A1 - Aloe-pig kidney compound composition and use thereof for manufacturing pharmaceuticals for treating kidney failure - Google Patents

Abstract

An aloe-pig kidney compound composition, comprising aloe extract dried powder and pig kidney powder. The aloe extract dried powder is obtained by concentrating decolorized aloe gel by 200:1, drying and grinding same into powder; and the pig kidney powder is obtained by drying and grinding the pig kidney into powder. The aloe pig kidney compound composition can effectively treat kidney failure and can be used for manufacturing pharmaceuticals for treating kidney failure.

Description

Aloe porcine kidney compound composition and use thereof for manufacturing medicine for treating kidney failure Technical field

The present invention relates to a composition (composition), particularly an aloe porcine kidney compound composition. The invention further relates to a use, in particular to the use of aloe vera porcine kidney composition for the manufacture of a medicament for the treatment of kidney failure.

Background technique

Renal failure, also referred to as renal failure, is mainly due to the incomplete renal function caused by kidney disease, which causes the kidney to fail to effectively remove metabolic waste when filtering blood. Renal failure can be divided into acute renal failure and chronic renal failure. The common symptoms of acute renal failure are oliguria (less than 400 ml of urine per day in adulthood), especially in patients with acute and severe disease. Complications, therefore, cause higher death; chronic renal failure refers to a decline in renal function to less than 60% for three months, and if it is chronically ill, it may cause irreversible kidney damage.

Aloe vera is a perennial herbaceous and succulent plant. In addition to its beauty effects such as moisturizing and post-sun repair, aloe vera extract is known to have antioxidant, anti-inflammatory, antibacterial and wound healing effects. In the prior art, for example, Chinese Patent Application Publication No. 104873677 discloses a traditional Chinese medicine vinegar for treating chronic renal failure, taking 8 parts of astragalus, 8 parts of cooked rhubarb, 8 parts of safflower, 8 parts of aloe, honey 8 Serve 60 parts of vinegar. Among them, Astragalus, cooked rhubarb and safflower are extracted by refluxing with ethanol, and then mixed with aloe vera, honey and vinegar to obtain a uniform. However, the traditional Chinese medicine vinegar combination lacks a scientific basis to confirm the efficacy, and there is still a lack of aloe-based compound for the treatment of renal failure.

In view of this, how to develop a composition for improving kidney failure, the prior art needs to be improved.

Summary of the invention

In order to overcome the disadvantages of the prior art, it is an object of the present invention to provide an aloe porcine kidney compound composition for achieving the efficacy of treating kidney failure.

In order to achieve the above object, the present invention provides an aloe porcine kidney compound composition comprising aloe vera extract dry powder and pig kidney powder; wherein the aloe extract dried powder is a decolorized aloe gel, concentrated at 200:1, and then dried and ground. In the form of powder, the pig kidney powder is dried and ground into powder.

Preferably, the weight ratio of the dried aloe extract powder to the porcine kidney powder is between 1:1 and 2:1, inclusive.

The present invention further provides a pharmaceutical for treating renal failure comprising an effective amount of the aloe porcine kidney compound composition according to claim 1 or 2 and a pharmaceutically acceptable carrier thereof.

Preferably, the pharmaceutical product is an enteral or parenteral dosage form.

More preferably, the enteral dosage form is an oral dosage form, and the oral dosage form is a solution, an emulsion, a suspension, a powder, a lozenge, a pill, an ingot, a tablet, a chewing gum or a capsule.

The present invention further provides a use of the aloe porcine kidney compound composition as described above for the manufacture of a medicament for treating renal failure, wherein the medicament comprises an effective amount of an aloe porcine kidney compound composition and a pharmaceutically acceptable carrier thereof.

The pharmaceutical of the present invention can be prepared into a dosage form suitable for the present invention by using the above-described aloe porcine kidney compound composition and a pharmaceutically acceptable carrier by a technique known to those skilled in the art. The "pharmaceutically acceptable carrier" as used in the present invention includes, but is not limited to, water, alcohols, glycols, hydrocarbons [such as petroleum jelly and white petrolatum). (white petrolatum)], wax (such as paraffin and yellow wax), preserving agents, antioxidants, solvents, emulsifiers, suspending agents (suspending agent), decomposer, binding agent, excipient, stabilizing agent, chelating agent, diluent, gelling agent Agent), preservative, lubricant, absorption enhancers, active agents, humectants, odor absorbers, fragrances, pH pH adjusting agents, occlusive agents, emollients, thickeners, solubilizing agents, penetration enhancers, anti-stimulation Anti-irritants, colorants, propellants, surfactants, and other carriers similar or suitable for use in the present invention.

Preferably, the effective dose of the aloe porcine kidney compound composition: aloe extract dry powder is between 0.13 g/g/kg/kg/day/day per kg, and the pig kidney powder is 0.13 g/kg/day to 0.4 g/kg/day, and the weight ratio of the aloe extract dry powder to the porcine kidney powder is between 1:1 and 2:1. The above dosages are calculated according to the Estimating the maximum safe starting dose in initial clinical trials for therapeutics in adult healthy volunteers.

More preferably, the effective dose of the aloe vera porcine kidney compound composition is: aloe vera extract dry powder is between 0.2 g/kg/day and 0.41 g/kg/day, and pig kidney powder is 0.2 g/kg/day. And the weight ratio of the aloe extract dry powder to the pig kidney powder is between 1:1 and 2:1.

Preferably, said renal failure, the pathway of its formation comprises regulation of fibrotic Smad (mothers against decapentaplegic) pathway protein expression (eg, phosphorylated-mothers against decapentaplegic homolog 2)/Smad 3 Protein expression and Smad 4 protein expression, inhibition of fibrosis pathway factor p-Smad 2/Smad 2 protein expression), promotion of inflammatory factors, reduction of total antioxidant capacity (TEAC), reduction of antioxidant enzyme activity or combination.

More preferably, the fibrotic pathway factor comprises, but is not limited to, α-smooth muscle actin (α-SMA), fibronectin (FN), and plasminogen activator inhibition. 1 (plasminogen activactor inhibitor-1, PAI-1), type 1 collagen (C-1), connective tissue growth factor (CTGF), TGF-β1.

More preferably, the inflammatory factor comprises, but is not limited to, transforming growth factor-β1 (TGF-β1), nuclear factor-κB (NF-κB), inducible oxidative Inducible nitric oxide synthase (iNOS), cyclooxygenase 2 (COX-2), interleukin-6 (IL-6), and tumor necrosis factor-α (tumor necrosis factor-α, TNF-α).

More preferably, the antioxidant enzymes include, but are not limited to, catalase, glutathione (GSH), glutathione peroxidase (GPx), superoxide. Superoxide dismutase (SOD) and glutathione reductase (GRd).

Preferably, the symptoms of renal failure comprise renal fibrosis.

The invention has the advantages that the aloe porcine kidney compound composition of the invention can improve renal failure, especially chronic renal fibrosis; the aloe porcine kidney compound composition initiates profibrosis by inhibiting the receptor on the TGF-β1 protein binding membrane. Smad pathway protein expression, inhibition of fibrosis pathway factors, inhibition of inflammatory factors to achieve anti-renal fibrosis; in addition, aloe vera porcine kidney compound composition also has the ability to enhance total antioxidant capacity and enhance the activity of antioxidant enzymes.

DRAWINGS

Figure 1 is a line drawing of the body weight of each group of experimental animals of the present invention; adenine is adenine; AV is an abbreviation for dry powder of aloe extract, and PK is an abbreviation for pig kidney powder. For details, please refer to Preparation Example 1, Figure 2 to Figure 17 have the same abbreviation.

Figure 2 is a photograph of the kidney appearance and kidney section of each group of experimental animals of the present invention; the top row is the kidney image taken by the mouse at the time of sacrifice; the second row is the Masson's trichrome stain, and the magnification is 100. The third row is hematoxylin and eosin stain (H&E stain), and the magnification is 100 times.

Figure 3 is a bar graph of the kidney weight of each group of experimental animals of the present invention; the statistics are mean ± standard deviation; ^## p < 0.01 is compared with the control group; ^* P < 0.05, ^** P < 0.01 is The induction group was compared.

4 is a protein electrophoresis pattern of TGF-β1 of kidney tissue of each group of experimental animals of the present invention; GAPDH (glyceraldehyde-3-phosphate dehydrogenase) is glyceraldehyde-3-phosphate dehydrogenase.

Figure 5 is a bar graph showing the expression level of TGF-β1 protein in kidney tissues of experimental animals of the present invention; the statistics are mean ± standard deviation; ^## p < 0.01 is compared with the control group; ^* P < 0.05, ^{* *} P < 0.01 compared to the induction group.

Figure 6 is a diagram showing the electrophoresis of p-Smad 2 and Smad 2 proteins in kidney tissues of experimental animals of the respective groups of the present invention.

/ Expression of the present invention. FIG. 7 animals in each experimental renal tissue p-Smad 2 Smad 2 protein in a bar chart; counted as mean ± standard deviation; ^## p <0.01 compared with the control group; ^** P < 0.01 was compared with the induction group.

Figure 8 is a diagram showing the electrophoresis of p-Smad 3 and Smad 3 proteins in kidney tissues of experimental animals of the respective groups of the present invention.

Figure 9 is a bar graph of the expression level of p-Smad 3/Smad 3 protein in kidney tissues of experimental animals of the present invention; the mean is ± standard deviation; ^## p < 0.01 is compared with the control group; ^* P <0.05, ^** P<0.01 was compared with the induction group.

Figure 10 is a diagram showing the Smad 4 protein electrophoresis pattern of kidney tissue of each group of experimental animals of the present invention.

Figure 11 is a bar graph showing the amount of Smad 4 protein expression in kidney tissues of experimental animals of the present invention; the statistics are mean ± standard deviation; ^## p < 0.01 is compared with the control group.

Figure 12 is a diagram showing the electrophoresis of α-SMA and FN proteins in kidney tissues of experimental animals of the respective groups of the present invention.

Figure 13 is a bar graph showing the expression levels of α-SMA and FN protein in kidney tissues of experimental animals of the present invention; the statistics are mean ± standard deviation; ^## p < 0.01 is compared with the control group; ^* P < 0.05 ^** P < 0.01 was compared with the induction group.

Figure 14 is a diagram showing the electrophoresis patterns of PAI-1, C-1 and CTGF proteins in kidney tissues of experimental animals of the respective groups of the present invention.

Figure 15 is a bar graph showing the expression levels of PAI-1, C-1 and CTGF proteins in kidney tissues of experimental animals of the present invention; the statistics are mean ± standard deviation; ^## p < 0.01 is compared with the control group; ^* P < 0.05, ^** P < 0.01 compared to the induction group.

Figure 16 is a diagram showing the electrophoresis of COX-2, NF-κB and iNOS proteins in kidney tissues of experimental animals of the respective groups of the present invention.

Figure 17 is a bar graph showing the expression levels of COX-2, NF-κB and iNOS protein in kidney tissues of experimental animals of the present invention; the statistics are mean ± standard deviation; ^# p < 0.05, ^## p < 0.01 is The control group was compared; ^* P<0.05, ^** P<0.01 was compared with the induction group.

detailed description

The technical means adopted by the present invention for achieving the intended purpose of the invention are further described below in conjunction with the drawings and preferred embodiments of the invention.

Preparation Example 1 Preparation of Aloe Vera Extract (Aloe vera)

The aloe extract used in the present invention is a dry extract powder (abbreviated as AV) of aloe extract extracted by a 200 kg of decolorized aloe vera gel provided by China Jiufulai Technology Group Co., Ltd. of China, and its product is referred to as AV. The name is "Aloe Vera Powder Capsule", the model is the fourth generation of 10% aloe polysaccharide-containing separation and preservation products; the extraction method of aloe extract dry powder is disclosed in Chinese Patent Application Publication No. 103554294, and used Peeled aloe vera for extraction.

Preparation 2 Preparation of pig kidney powder

The pig kidneys purchased from the market are washed and cut into thick slices and boiled at 100 ° C for 5 minutes, then rinsed in a freezer at -80 ° C for 1 day, then taken out for freeze drying, until the pig kidney is thoroughly dehydrated and dried. Grinding into a powder by a homogenizer, that is, obtaining pig kidney powder (PK for short), and storing it in a dry box for storage.

Example 1 Effect of aloe porcine kidney compound composition on body weight changes of adenine induced mice

After 6 weeks old male C57BL/6 mice were placed in the room, the feeding conditions were automatic air conditioning (air exchange rate 12 times per hour), automatic light control (12 hours daylight, 12 hours dark night), average room temperature 22±2 ° C, relative Humidity 50% to 55%, free to eat general feed (Taiwan Lesco Biotechnology Co., Ltd. purchase number MFG22, as shown in Table 1 below) and drinking water.

Table 1, MFG22 feed composition

ingredient	content		ingredient	content
Vitamin A	2900IU/100g		Moisture	7.9%
Vitamin D3	650 IU/100g		Crude Protein	22.9%
Vitamin E	21.2mg/100g		Crude Fat	5.4%
Vitamin B1	4.4mg/100g		Crude Ash	6.2%
Vitamin B2	3.01mg/100g		Crude Fiber	3.4%
Vitamin B6	1.05mg/100g		Nitrogen free extract	54.2%
			Calories	357 kcal / 100 g

After the animals were acclimated to the environment for 7 days, they were randomly assigned to six groups (8/group):

(1) Control group (Control): standard feed, normal diet.

(2) Induction group (adenine 0.2%): The standard feed was added in an amount of 0.2% (w/w) adenine.

(3) Low-dose aloe vera combined with pig kidney test group (adenine 0.2%+AV 1%+PK 1%): 0.2% (w/w) adenine was added to standard feed, and 1% (w/w) of preparation example 1 was taken. Aloe extract dry powder (AV 1%), and 1% (w/w) pig kidney powder of Preparation Example 2 (PK 1%).

(4) High-dose aloe vera combined with pig kidney test group (adenine 0.2%+AV 2%+PK 1%): 0.2% (w/w) adenine was added to standard feed, and 2% (w/w) of preparation example 1 was taken. Aloe extract dry powder (AV 2%) and 1% (w/w) pig kidney powder of Preparation Example 2 (PK 1%).

(5) Aloe vera test group alone (adenine 0.2%+AV 2%): 0.2% (w/w) adenine was added to standard feed and 2% (w/w) aloe extract dry powder of preparation example 1 (AV 2) %).

(6) Individual pig kidney test group (adenine 0.2% + PK 1%): Standard feed #MFG22 was supplemented with 0.2% (w/w) adenine and 1% (w/w) pig kidney powder (PK 1%).

During the experiment, each mouse consumed an average of about 5 grams per day, and the diet contained all but the control group. 0.2% (w/w) adenine (ie 0.01 g adenine/day/day) and AV 1% (0.05 g AV/day/day), AV 2% (0.1 g AV/day/day) or PK 1% (0.05 g PK/day/day) was mixed into the powdered feed at the doses of the above different groups to prepare a fed mice. Except for the control group, after feeding for 0.2% (w/w) adenine every day for 4 weeks, AV and PK were mixed in standard diet for 6 weeks. Weekly monitoring of body weight during the experiment continued until the end of the experiment at week 10, and the body weight of the rats was measured and recorded on Tuesday.

Table 2. Body weight values for the 2nd to 10th week of each group of mice (g)

Note: The statistics are mean±standard deviation (n=8); ^## p<0.01 is compared with the control group; ^* P<0.05, ^** P<0.01 is compared with the induction group.

The body weight values are shown in Figure 1 and Table 2 above. The body weight of the induction group was significantly decreased after feeding adenine. The induction group decreased from 0.81, 0.72, 0.67, and 0.67 in the control group to the control group from the fifth week to the tenth week. 0.7, 0.68 times. After the end of the fourth week, after feeding different doses of the therapeutic drug, the body weight value showed that the adenine 0.2%+AV 2%+PK 1% test group increased significantly at the 6th to 10th week, and increased by 1.18 compared with the induction group. 1.16, 1.13, 1.1 and 1.15 times. There was no difference between the adenine 0.2%+AV 1%+PK 1% test group and the induction group. The adenine 0.2%+AV 2% test group increased by 0.92 and 1.1 fold, respectively, at weeks 5 and 10 compared with the induction group. The adenine 0.2%+PK 1% test group increased by 0.89, 0.92, and 0.93 times at 5 weeks and 7 to 8 weeks, respectively, compared with the induction group. Therefore, the aloe porcine kidney compound composition has the effect of restoring body weight and has a synergistic effect at weeks 6 to 9.

Example 2 Effect of aloe porcine kidney compound composition on serum biochemical changes in mice induced by adenine

The group was the same as in Example 1, except that the control group was fed with 0.2% (w/w) adenine every day for 4 weeks, and AV and/or PK were separately mixed in standard feed for 6 weeks. The experiment was carried out for a total of 10 weeks. At the end of the experiment, the animals were sacrificed and the blood was allowed to stand for 30 minutes. The cells were centrifuged at 3000 rpm, 4 ° C, and 15 minutes using an ultra-high speed centrifuge, and the supernatant and serum were extracted. experiment analysis.

Table 3, serum biochemical values

Note: The statistics are mean ± standard deviation (n=8); ^## p<0.01 is compared with the control group; ^* P<0.05, ^** P<0.01 compared with the induction group; BUM (blood urea nitrogen ) is blood urea nitrogen; CRE (creatinine) is creatinine; BS (blood sugar) is blood sugar; GOT (glutamic oxaloacetic transaminase) is glutamic acid oxaloacetate transaminase; GPT (glutamic pyruvic transaminase) is glutamic acid pyruvate Transaminase; TCHO (total cholesterol) is total cholesterol; TG (triglyceride) is triglyceride.

The results are shown in Table 3 above. BUN and CRE results in renal function evaluation showed that BUN and CRE increased in the induction group after feeding adenine, which was 3.47 and 2.7 times higher than that in the control group, respectively, and both showed significant differences in adenine. 0.2%+AV 1%+PK 1% or adenine 0.2%+AV 2%+PK 1% The test group had a decreasing trend in BUN and CRE compared with the induction group, while adenine 0.2%+AV 2% test group and There was no significant difference in BUN and CRE between the adenine 0.2%+PK 1% test group and the induction group. In terms of blood glucose evaluation, the induction group did not reach statistical difference after comparison with the control group, but the adenine 0.2%+AV 2% test group or the adenine 0.2%+PK 1% test group increased significantly by 1.41 compared with the induction group. , 1.36 times. In the liver functional assessment GOT and GPT results showed that the induction group compared with the control group, or different doses of AV and / or PK test group compared with the induction group, did not reach statistical differences. TCHO and TG were evaluated in lipid metabolism. Compared with the control group, TCHO increased by 1.28 times and TG decreased. In the adenine 0.2%+AV 1%+PK 1% test group, TCHO was significant compared with the induction group. 1.1 times higher, TG did not reach significant difference, in adenine 0.2% + AV 2% + PK 1% test group, adenine 0.2% + AV 2% test group or adenine 0.2% + PK 1% test group, in TCHO and TG There are no statistically significant differences. Therefore, the aloe porcine kidney compound composition can reduce BUN and have a multiplication effect.

Example 3 Effect of Aloe Pig and Kidney Compound Composition on the Level of Antioxidant Enzyme and Lipid Peroxidation in Kidney of Mice Induced by Adenine

The group was the same as in Example 1, except that the control group was fed with 0.2% (w/w) adenine every day for 4 weeks, and AV and/or PK were separately mixed in standard feed for 6 weeks. The experiment was carried out for a total of 10 weeks, and the mouse kidney tissue was taken for analysis after the end of the experiment.

Table 4. Renal antioxidant enzyme activity and lipid peroxidation

Note: The statistics are mean ± standard deviation (n=8); ^# p<0.05, ^## p<0.01 compared with the control group; ^* P<0.05, ^** P<0.01 compared with the induction group; TEAC Activity (trolox equivalent antioxidant capacity) is the total antioxidant capacity (nmol / protein mg); catalase activity is catalase activity (hydrogen peroxide consumption μmol / protein mg); MDA (malondialdehyde) concentration is malondialdehyde concentration ( Nmol/protein mg); GSH (glutathione) concentration is glutathione concentration (μM/protein mg); GPx (glutathione peroxidase) activity is glutathione peroxidase activity [oxidized nicotinic adenine dinuclear nucleus Glucuronide phosphate (NADPH) nmol/min/protein mg]; SOD (superoxide dismutase) activity is superoxide dismutase activity (U/ml/protein mg); GRd (glutathione reductase) activity is glutathione reductase activity (NADPH nmol/min/ml/protein mg); IL-1β (interleukin-1β) is interleukin-1β (pg/ml); IL-6 (interleukin-6) is interleukin-6 (pg/ml); TNF- α (tumor necrosis factor-α) is tumor necrosis factor-α (pg/ml).

The results are shown in Table 4 above. Compared with the control group, the antioxidant group can significantly reduce the antioxidant enzymes such as: catalase, GSH, SOD and GRd to 0.77, 0.47, 0.48 and 0.74 times of the control group, respectively. There is no significant difference between TEAC and GPx. Compared with the induction group, the adenine 0.2%+AV 1%+PK 1% test group significantly increased the antioxidant enzymes such as TEAC, catalase, GSH, SOD and GRd by 1.19, 1.71, 3.04, 1.3 and 1.21 times, respectively. However, there is no statistically significant difference in GPx. Compared with the induction group, the AV 2%+PK 1% test group significantly increased TEAC, catalase, GSH, GPx, SOD and GRd, which increased by 1.09, 1.57, 1.65, 1.63, 1.46 and 1.25 times, respectively. Compared with the induction group, the adenine 0.2%+AV 2% test group significantly increased TEAC, catalase, GSH, GPx, SOD and GRd by 1.12, 1.58, 2.4, 1.84, 1.49 and 1.4 times, respectively. Compared with the induction group, GSH and GRd increased significantly by 2.04 and 1.34 times in the adenine 0.2%+PK 1% test group, but increased in GPx and SOD but did not reach significant difference, while TEAC and catalase had no statistics. Significant differences.

The thiobarbituric acid reactive substances (TBARs) were mainly analyzed by the method established by the scholar Ohkawa in 1979. Please refer to Table 4 above. The MDA production of lipid peroxidation index is compared between the induction group and the control group. Significantly increased by 1.16 times, while adenine 0.2%+AV 1%+PK 1% test group or adenine 0.2%+AV 2%+PK 1% test group was reduced to 0.58 and 0.67 times MDA in the induction group, respectively, compared with the induction group. There was no statistically significant difference in the adenine 0.2%+AV 2% test group or the adenine 0.2%+PK 1% test group compared with the induction group. Therefore, the aloe porcine kidney compound composition can significantly reduce the degree of lipid peroxidation.

Example 4 Effect of aloe porcine kidney compound composition on adenine-induced renal inflammatory factors in mice

The group was the same as in Example 1, except that the control group was fed with 0.2% (w/w) adenine every day for 4 weeks, and AV and/or PK were separately mixed in standard feed for 6 weeks. The experiment was carried out for a total of 10 weeks, and the mouse kidney tissue was taken for analysis after the end of the experiment.

Table 5. Determination of renal cytokine levels

Note: The statistics are mean ± standard deviation (n=8); ^# p<0.05, ^## p<0.01 compared with the control group; ^* P<0.05, ^** P<0.01 compared with the induction group; IL -1β(interleukin-1β) is interleukin-1β (pg/ml); IL-6 (interleukin-6) is interleukin-6 (pg/ml); TNF-α (tumor necrosis factor-α) is tumor necrosis factor- α (pg/ml).

The results are shown in Table 5 above. Compared with the control group, the induction group can increase the expression of IL-6 and TNF-α, which are increased by 3.04 and 1.98 times, respectively. Statistically, the increase of IL-6 is most significant in IL-1β. Although there is an increasing trend, it has not reached significant. The combined treatment group, adenine 0.2%+AV 1%+PK 1% test group or adenine 0.2%+AV 2%+PK 1% test group, IL-6 can be reduced by 0.34, 0.41 compared with the induction group.倍, TNF-α can be reduced to 0.56, 0.5 times of the induction group, respectively, in IL-1β only adenine 0.2% + AV 2% + PK 1% test group has a decreasing trend and has a multiplication effect. Compared with the induction group, the adenine 0.2%+AV 2% test group reduced the IL-6 and TNF-α by 0.28 and 0.45 times, respectively; in the adenine 0.2%+PK 1% test group compared with the induction group. , can reduce the expression of IL-6 and TNF-α in the kidneys of 0.24 and 0.55 times of the induction group, respectively. From the above results, it can be known that AV and PK have the ability to reduce the inflammatory response whether it is a combination or a separate treatment.

Example 5 Effect of aloe porcine kidney compound composition on adenine-induced kidney weight and pathological section in mice

The group was the same as in Example 1, except that the control group was fed with 0.2% (w/w) adenine every day for 4 weeks, and AV and/or PK were separately mixed in standard feed for 6 weeks. The experiment was carried out for a total of 10 weeks, and the mouse kidney tissue was taken for analysis after the end of the experiment. After the sacrifice of the mice, the kidneys were subjected to hematoxylin-eosin staining and fibrosis staining. The kidney stained with the kidney spheroids and photographed at a magnification of 100 times. The pathological features of the kidneys were observed under the microscope.

Please refer to Figure 2, the top row is the kidney image taken by the mouse at the moment of sacrifice. It can be observed from the photo that the color of the kidney in the control group is more rosy and shiny, which is a characteristic of healthy kidney. Compared with the control group, the induction group showed no ruddy color and obvious protruding particles on the surface of the whole kidney surface, and the touch was rough. In the adenine 0.2%+AV 1%+PK 1% test group or adenine 0.2%+AV 2%+PK 1% test group, compared with the induction group, the surface of the kidney has less protruding particles, and the body touch is more delicate and shiny. The color part was more rosy and shiny with adenine 0.2%+AV 2%+PK 1%. In the adenine 0.2%+AV 2% test group or the adenine 0.2%+PK 1% test group, the body touch was also smoother than the induction group. According to the results of kidney quality analysis in Figure 3, the induction group was 0.37 times smaller than the control group, and there was significant atrophy, while adenine 0.2%+AV 1%+PK 1% test group or adenine 0.2%+AV 2%+ Compared with the induction group, the PK 1% test group increased the kidney mass by 1.65 and 1.5 times, respectively, while the adenine 0.2%+AV 2% test group or the adenine 0.2%+PK 1% test group increased compared with the induction group. the trend of.

Referring to Figure 2, the second row is fibrillated. If the tissue is fibrotic, the staining results will appear blue. The results showed that there was no fibrosis in the control group and significant fibrosis in the induction group. In adenine 0.2%+AV 1%+PK 1% test group, adenine 0.2%+AV 2%+PK 1% test group, adenine 0.2%+AV 2% test group, or adenine 0.2%+PK 1% test group Compared with the induction group, the fibrosis phenomenon was reduced, and the effect of aloe vera pig kidney compound composition on improving kidney damage was obvious.

Referring to Figure 2, the results of the third row of hematoxylin-eosin staining showed that the saccular space in the renal spheroid of the control group was small, and the epithelial cells in the lumen of the surrounding renal tubules were regular and dense. The cell type is tight and full, and the space in the lumen is clear without any metabolite deposition. The number of renal tubules in the 100-fold field of view is dense and complete as a whole, which is a pathological feature of healthy renal tubular epithelial cells. Compared with the control group, the induction group can be found that feeding adenine will cause the saccular space in the renal spheroid to become larger, and the number of renal tubular epithelial cells distributed around the renal spheroid is reduced and the shape is flat, making the renal tubule The diameter of the tube is enlarged or even disintegrated, and the adenine metabolite 2,8-dihydroxyadenine yellow crystal deposit can be observed in the lumen, and the distribution of renal tubules in the 100-fold field of view is also significantly sparse. The adenine 0.2%+AV 1%+PK 1% test group or adenine 0.2%+AV 2%+PK 1% test group compared with the induction group, the renal tubular distribution in the 100-fold field of view was significantly increased, and the cell type was full. The arrangement of luminal epithelial cells was also dense, and the deposition of 2,8-dihydroxyadenine yellow crystals was relatively reduced, but there was no significant difference in the interstitial saccular space in the glomeruli. Compared with the induction group, the adenine 0.2%+PK 1% test group can significantly improve the pathological features of renal injury, while the adenine 0.2%+AV 2% test group has no significant difference compared with the induction group. Therefore, the aloe porcine kidney compound composition can significantly improve the pathological features of kidney injury.

Example 6 Effect of aloe porcine kidney compound composition on the expression of fibrin TGF-β1 in the kidney of mice induced by adenine

The group was the same as in Example 1, except that the control group was fed with 0.2% (w/w) adenine every day for 4 weeks, and AV and/or PK were separately mixed in standard feed for 6 weeks. The experiment was carried out for a total of 10 weeks. At the end of the experiment, the kidney tissue of the mice was taken and analyzed by western blotting analysis.

From the results of Fig. 4 and Fig. 5, the induction group can enhance the expression of TGF-β1 protein, which is 8.48 times higher than that of the control group. Adenine 0.2%+AV 1%+PK 1% test group or adenine 0.2%+AV 2%+PK 1% test group can significantly reduce TGF-β1 protein expression 0.43, 0.32 times of induction group, respectively, compared with the induction group. Among them, adenine 0.2%+AV 2%+PK 1% test group reduced TGF-β1 protein expression most significantly, while adenine 0.2%+AV 2% test group or adenine 0.2%+PK 1% test group had decreased TGF-β1 The trend of protein expression. Therefore, the aloe porcine kidney compound composition can significantly reduce the expression of fibrosis-related proteins.

Example 7 Effect of aloe porcine kidney compound composition on the expression of Smad transmission pathway-related proteins in adenine-induced mouse kidney

The group was the same as in Example 1, except that the control group was fed with 0.2% (w/w) adenine every day for 4 weeks, and AV and/or PK were separately mixed in standard feed for 6 weeks. The experiment was carried out for a total of 10 weeks. At the end of the experiment, the kidney tissue of the mice was taken and analyzed by Western blotting method. The expression of profibrotic Smad pathway protein was initiated by the receptor on the TGF-β1 protein binding membrane.

The results of Fig. 6 and Fig. 7 show that the expression ratio of p-Smad2/Smad 2 protein is significantly higher in the induction group than in the control group. The decrease was 0.63 times that of the control group, and was significantly increased by 1.83 times in the adenine 0.2%+AV 2%+PK 1% test group, and had a multiplication effect. There was no statistically significant difference in the adenine 0.2%+AV 1%+PK 1% test group, the adenine 0.2%+AV 2% test group, or the adenine 0.2%+PK 1% test group compared with the induction group. The results in Figure 8 and Figure 9 show that the expression ratio of p-Smad 3/Smad 3 protein increased significantly by 1.77 times in the induction group compared with the control group, in the adenine 0.2%+AV 1%+PK 1% test group, adenine 0.2%+ The AV 2%+PK 1% test group, the adenine 0.2%+AV 2% test group, or the adenine 0.2%+PK 1% test group significantly reduced the p-Smad 3/Smad 3 protein expression ratio compared with the induction group. The reduction was 0.29, 0.42, 0.52, and 0.35 times of the induction group, respectively, and the test group was most significant with adenine 0.2%+AV 1%+PK 1%. Figures 10 and 11 show that the induction group showed a significant 1.55 fold increase in Smad 4 protein expression compared to the control group. In adenine 0.2%+AV 1%+PK 1% test group, adenine 0.2%+AV 2%+PK 1% test group, adenine 0.2%+AV 2% test group, or adenine 0.2%+PK 1% test group There were no statistical differences in the induction group. Therefore, the aloe porcine kidney compound composition significantly increased the p-Smad2/Smad 2 protein expression ratio and significantly reduced the p-Smad 3/Smad 3 protein expression ratio.

Example 8 Effect of Aloe Pig and Kidney Compound Composition on Expression of Fibrosis Pathway Related Protein in Kidney of Mice Induced by Adenine

The group was the same as in Example 1, except that the control group was fed with 0.2% (w/w) adenine every day for 4 weeks, and AV and/or PK were separately mixed in standard feed for 6 weeks. The experiment was carried out for 10 weeks. At the end of the experiment, the kidney tissue of the mice was taken and analyzed by Western blotting method to analyze the expression of the pathway protein of the epithelial mesenchymal transition (EMT) downstream of the Smad signaling pathway.

The results of Fig. 12 and Fig. 13 show that the expression of α-SMA and FN protein is significantly increased in the induction group compared with the control group, which is increased by 29.62 and 27 times, respectively. Adenine 0.2%+AV 1%+PK 1% test group or adenine 0.2%+AV 2%+PK 1% test group can significantly reduce α-SMA protein expression compared with the induction group, respectively, reduced to 0.5 in the induction group. 0.27 times, in the adenine 0.2% + AV 2% test group or adenine 0.2% + PK 1% test group, there is a tendency to reduce the expression of α-SMA protein. In the FN protein expression, the induction group was significantly increased by 27.07 times compared with the control group, while the adenine 0.2%+AV 1%+PK 1% test group, adenine 0.2%+AV 2%+PK 1% test group, and adenine 0.2%+ In the PK 1% test group, the expression of FN protein was significantly decreased to 0.6, 0.56, and 0.42 times in the induction group, respectively. In the adenine 0.2%+AV 2% test group, the expression of FN protein decreased. Therefore, the aloe porcine kidney compound composition can significantly reduce the expression of α-SMA and FN protein and have a multiplication effect.

The EMT process is then analyzed to convert increased myofibroblasts, which are associated protein expression for the extracellular matrix (ECM) accumulation in the renal interstitial region. The results of Fig. 14 and Fig. 15 showed that the induction group increased the expression of extracellular matrix proteins PAI-1, C-1 and CTGF, which increased by 24.83, 40.25 and 16.38 times, respectively. Compared with the induction group, the adenine 0.2%+AV 1%+PK 1% test group Significantly reduced PAI-1 and C-1 protein expression were reduced to 0.62 and 0.56 times in the induction group, respectively, while CTGF showed a downward trend, but did not reach statistical difference. Compared with the induction group, the high-dose AV combined with PK test group and the induced group can significantly reduce the expression of PAI-1, C-1 and CTGF protein in the adenine 0.2%+AV 2%+PK 1% test group, respectively, and reduced to the induction group. 0.49, 0.15, 0.24 times. In the adenine 0.2%+AV 2% test group, the C-1 protein expression was significantly reduced to 0.43 times in the induction group, and the adenine 0.2%+PK 1% test group reduced the C-1 protein expression to 0.7 times in the induction group. trend. Therefore, the aloe porcine kidney compound composition can significantly reduce the expression levels of PAI-1, C-1 and CTGF proteins and have a multiplication effect.

Example 9 Effect of aloe porcine kidney compound composition on the expression of inflammation-related proteins in the kidney of mice induced by adenine

The group was the same as in Example 1, except that the control group was fed with 0.2% (w/w) adenine every day for 4 weeks, and AV and/or PK were separately mixed in standard feed for 6 weeks. The experiment was carried out for a total of 10 weeks. At the end of the experiment, the kidney tissue of the mice was taken and the results of the inflammation-related proteins were analyzed by Western blotting.

The results of Fig. 16 and Fig. 17 showed that the protein expressions of inflammatory related proteins COX-2, NF-κB and iNOS were increased by 7.56, 4.34 and 4.49 times, respectively, compared with the control group, and all reached statistically significant differences. In the experimental group, the protein expression of NF-κB and iNOS increased by 1.41 and 1.28 times, respectively, in the adenine 0.2%+AV 1%+PK 1% test group compared with the induction group, and the increase of NF-κB was significantly different. Compared with the induction group, the adenine 0.2%+AV 2%+PK1% test group significantly reduced the expression of proinflammatory cytokines COX-2, NF-κB and iNOS, which were reduced to 0.5, 0.74, and 0.2 in the induction group, respectively. Times, the statistical difference is significant. Compared with the induction group, the expression of iNOS protein in adenine 0.2%+AV 2% test group was significantly reduced to 0.38 times that of the induction group. There was no statistical difference in COX-2 and NF-κB protein expression. Compared with the induction group, the adenine 0.2%+PK 1% test group showed a significant increase in COX-2 protein expression, an increase of 1.05 times. Although there was an increasing trend in NF-κB and iNOS, it did not reach statistically significant difference. Therefore, the aloe porcine kidney compound composition can significantly reduce the expression of COX-2, NF-κB and iNOS protein and have a multiplication effect.

The above is only a preferred embodiment of the present invention, and is not intended to limit the scope of the present invention. Although the present invention has been described above by way of preferred embodiments, it is not intended to limit the invention. Those skilled in the art can make some modifications or modifications to equivalent embodiments by using the above-disclosed technical contents without departing from the technical scope of the present invention. It is still within the scope of the technical solution of the present invention to make any simple modifications, equivalent changes and modifications to the above embodiments.

Claims (9)

1. An aloe porcine kidney compound composition, characterized in that the aloe porcine kidney compound composition comprises aloe vera extract dry powder and pig kidney powder; wherein the aloe extract dry powder is a decolorized aloe vera gel and concentrated at 200:1 and dried. Grinded into powder, the pig kidney powder is dried and ground into powder.
2. The aloe porcine kidney compound composition according to claim 1, wherein the weight ratio of the aloe extract dry powder to the porcine kidney powder is between 1:1 and 2:1.
3. A pharmaceutical product for treating renal failure, characterized in that the pharmaceutical product comprises an effective amount of the aloe porcine kidney compound composition according to claim 1 or 2 and a pharmaceutically acceptable carrier thereof.
4. The pharmaceutical according to claim 3, wherein the pharmaceutical product is an enteral or parenteral dosage form.
5. The pharmaceutical according to claim 4, wherein the enteral dosage form is an oral dosage form, and the oral dosage form is a solution, an emulsion, a suspension, a powder, a tablet, a pill, an ingot, a tablet, Chewing gum or capsules.
6. The use of the aloe porcine kidney compound composition according to claim 1 or 2 for the manufacture of a medicament for treating renal failure, wherein the medicament comprises an effective dose of aloe vera porcine kidney compound composition and a pharmaceutically acceptable load thereof Agent.
7. The use according to claim 6, wherein the effective dose of the aloe porcine kidney compound composition: aloe vera extract dry powder is between 0.13 g (g/kg/day) to 0.81 g/kg/kg/day Kg/day, pig kidney powder is from 0.13 g/kg/day to 0.4 g/kg/day, and the weight ratio of aloe extract dry powder to pig kidney powder is between 1:1 and 2:1.
8. The use according to claim 6, characterized in that the renal failure pathway comprises a pathway for promoting fibrotic-mothers against decapentaplegic homolog 2/Smad 3 protein and inhibiting fibrosis pathway Factor p-Smad 2/Smad 2 protein expression, promotion of inflammatory factors, reduction of total antioxidant capacity (TEAC), reduction of antioxidant enzyme activity or a combination thereof.
9. Use according to any one of claims 6 to 8 wherein the symptoms of renal failure comprise renal fibrosis.

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