Effects of bisdemethoxycurcumin in reducing lipids and fighting lipid peroxidation
`
`Xingqiu Hong *, Yanfen Huang, Minmin Fu, Hailong Liu
`
` Zhejiang Chinese Medical University, Life Science Department, Hangzhou 310053
`
`[Abstract] Purpose: To investigate the effects of bisdemethoxy curcumin (Bdmc) in reducing lipids
`and fighting lipid peroxidation, and its influence on the activity of enzymes associated with lipid
`metabolism. Method: Wistar rats were fed a high-lipid diet, and after creating diet-induced
`hyperlipidemia, bisdemethoxycurcumin and the positive control drug simvastatin were used to conduct
`experimental treatment. After 3 weeks of drug administration, the animals were killed, serum and liver
`total cholesterol (TC) and triglycerides (TG) and serum high-density lipoprotein (HDL-C) content were
`measured, low-density lipoprotein (LDL-C) content was calculated, and post-heparin lipolytic activity
`(PHLA) and the activity of lipoprotein lipase (LPL) and hepatic lipase (HL) were measured, as well as serum
`malondialdehyde (MDA) content. Results: Both bisdemethoxycurcumin and simvastatin can lower serum
`and
`liver TC and TG content, elevate serum HDL-C content, and
`lower LDL-C content.
`Bisdemethoxycurcumin can significantly elevate PHLA activity, and its effect of elevating LPL and HL
`activity is also notable; bisdemethoxycurcumin can effectively reduce MDA content. Conclusion:
`Bisdemethoxycurcumin has the effects of reducing liver and serum lipids, and in particular, it has the
`effects of notably lowering TG and elevating the activity of enzymes associated with TG metabolism.
`
` [Key words] bisdemethoxycurcumin; hyperlipidemia; lipoprotein lipase; hepatic lipase
`
` [Chinese library classification number] R965.1 [Document code] A [Article number] 1672-3651
`(2006) 02-0121-04
`
`Hyperlipidemia, particularly oxidized lipoprotein, is one of the most dangerous factors leading to
`atherosclerosis (As) and cardiovascular and cerebrovascular diseases. Therefore, effectively lowering high
`plasma lipoprotein levels and controlling the formation of oxidized lipoproteins can reduce the occurrence
`and development of cardiovascular and cerebrovascular diseases. Turmeric is a common traditional
`Chinese medicine in China; it has the effect of promoting blood circulation and removing stasis. Having
`undergone modern pharmacological studies, turmerol extract and its main ingredient curcumin have
`functions such as reducing lipids and fighting lipid peroxidation [1, 2], and they have been clinically applied
`as five types of drugs. The bisdemethoxycurcumin ingredient of turmerol extract can inhibit the
`proliferation of smooth muscle cells and endothelial cells proliferation [3, 4], but there have been no reports
`on its effect on animal lipids, lipid peroxidation, and the activity of enzymes associated with lipid
`metabolism, or whether it is the main active ingredient of turmerol extract.
`
`1 Materials and methods
`
`1.1 Drugs
`
`turmerol extract via silica gel column
`from
`Bisdemethoxycurcumin was separated
`recrystallization, was analyzed and
`chromatography, underwent CHCl3
`compared
`to a
`
`SAB1008
`U.S. Pat. No. 10,945,970
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`

`

`bisdemethoxycurcumin reference substance (Chengdu Tianyin Industrial Co., Ltd.), and was found to be
`the same substance. Using the spectrophotometric method [5] and measuring at 505 nm, its purity was
`95.3%. Simvastatin was produced by Harbin Pharmaceutical Group Sanjing Pharmaceutical Co., Ltd.
`
`1.2 Establishment and grouping of diet-induced hyperlipidemia rat models
`
`49 male Wistar rats weighing 150 g - 170 g were provided by the Shanghai Laboratory Animal
`Center of the Chinese Academy of Sciences. The conditions under which they were raised were 5 animals
`per cage, at a temperature of 23±1°C and a relative humidity of 40% - 70%. After being allowed to
`acclimate for one week, 8 animals were randomly selected to be the normal control group and were given
`basal feed (Qi leather powder 25%, bean flour 15%, fish meal 10%, corn flour 20%, no. 4 powder 25%, salt
`1%, cod liver oil 1%, and other 3%). The rest were fed a high-lipid diet (cholesterol 2%, lard 10%, cholate
`0.25%, and basal feed 87.75%). They were given free access to their food, and their blood lipids were
`measured after 2 weeks. The animals were ranked according to blood lipid levels and divided into 5 groups
`(with 9 animals in the model group). While continuing to administer the high-lipid diet, the model group
`was given 5 mL  kg-1  d-1 1% carboxymethyl cellulose by gavage, the positive control group was given
`simvastatin 0.012 mmol  kg-1  d-1, the high-dosage bisdemethoxycurcumin group was given 0.3 mmol 
`kg-1  d-1, the medium-dosage group was given 0.1 mmol  kg-1  d-1, and the low-dosage group was given
`0.03 mmol  kg-1  d-1. Gavage was performed after homogenization with 5 mL 1% carboxymethyl cellulose
`sodium solution for every kg used. 5 mL  kg-1 was administered to the normal group by gavage every day,
`the animals were weighed every other week, and the dosages were promptly adjusted according to weight.
`The animals were killed after three weeks of drug administration. Food was withheld starting at 8PM two
`days before they were killed, orbital blood collection was conducted in the morning one day before they
`were killed, and the serum was used to measure TC, TG, and MDA. Feeding was resumed following blood
`collection, and feed was again withheld at 8PM that night. In the morning on the day the animals were
`
`[Draft received] 2005-08-11
`
`[Funding] Funding project of the Zhejiang Province Natural Science Foundation (No. 202015)
`
`[*Corresponding author] Hong Xingqiu: Associate research fellow, master’s student advisor, Tel.: 0571-
`86613724, E-mail: xqhong20204@163.com
`
`killed, heparin 312.5 μg  kg-1 was injected into the caudal vein, and after 15 min, the eyes were removed
`and heparinized plasma was collected, for measuring PHLA, LPL, and HL activity. After the blood was
`collected, the rats were killed by breaking their necks, and their livers were removed to measure liver TC
`and TG.
`
`1.3 Measurement methods
`
`Serum TC and TG were measured using the enzyme method, and HDL-C was measured by
`removing LDL and VLDL using the phosphotungstic acid – magnesium precipitation method and then
`measuring the upper layer using the enzyme method. The reagent kits were produced by Ningbo Cicheng
`Biochemical Reagent Factory. LDL-C content was calculated using the formula LDL-C = TC – HDL-C – 1/5TG.
`For liver TC and TG measurements, normal saline was added to the samples, centrifugation was performed
`after homogenization [6], and reagent kits were used as described above to take the measurements. PHLA,
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`HL, and LPL activity were measured using LPL and HL reagent kits produced by Nanjing Jiancheng
`Biochemical Reagent Factory. 2x20 μL of heparinized plasma were taken, and using a lipid emulsion as the
`substrate, different enzyme-specific inhibitors were added separately. The emulsions were mixed and left
`for a certain amount of time at 37°C. The LPL and HL in the plasma were able to hydrolyze the TG in the
`substrate as glycerol and free fatty acids (FFA), respectively. Copper and chromogenic reagents were
`added, a spectrophotometer was used to measure the generated FFA content at 550 nm, and LPL and HL
`activity were separated calculated. Adding the two gives PHLA activity.
`
`1.4 MDA measurement
`
`MDA was measured using MDA reagent kits; the reagent kits were produced by Nanjing Jiancheng
`Biochemical Reagent Factory. Lipid peroxide degradation can produce MDA, which condenses with
`thiobarbituric acid, forming a red product. Absorbance was measured at 532 nm, and MDA content was
`calculated.
`
`1.5 Statistical processing
`
`The data obtained was expressed as
`groups.
`
`2 Results
`
`, and a t test was employed to compare the two
`
`2.1 Influence of bisdemethoxycurcumin on rat serum TC, TG, HDL-C, and LDL-C content
`
`Both simvastatin and high-dose bisdemethoxycurcumin can lower serum TC and elevate HDL-C
`content (but at the doses used in the experiment, the simvastatin had the smallest TC average); in addition,
`simvastatin and high- and medium-dose bisdemethoxycurcumin all can lower serum TG and LDL-C content,
`wherein
`the high-dosage bisdemethoxycurcumin group had
`the best
`results;
`low-dose
`bisdemethoxycurcumin has no notable effect on TC and TG (see Table 1 for results).
`
`2.2 Influence of bisdemethoxycurcumin on rat liver TC and TG content
`
`Both bisdemethoxycurcumin and simvastatin can notably lower liver TC and TG content, similarly
`to the effect of lowering TC and TG in serum. That is, the effect of bisdemethoxycurcumin on lowering TG
`is quite strong, and there are significant differences between the high-, medium-, and low-dose groups
`(see Table 2 for results).
`
`2.3 Influence of bisdemethoxycurcumin on enzyme activity
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`Comparing the heparinized plasma lipolytic enzyme, lipoprotein lipase, and hepatic lipase activity
`
`of the various groups to the high-lipid model group, Table 3 indicates that there is a significant reduction
`in PHLA, HL, and LPL activity in the model group, the simvastatin has a significant activating effect on PHLA,
`and the bisdemethoxycurcumin high-dose group has a significant elevation of PHLA, HL, and LPL activity.
`
`2.4 Influence of bisdemethoxycurcumin on serum MDA content
`
`The high-lipid model group had notably elevated MDA content, bisdemethoxycurcumin can
`
`significantly lower MDA content, and the simvastatin group had no notable influence on MDA content
`(see Table 4).
`
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`3 Discussion
`
`
`
`Reducing lipids is a complicated process involving multiple aspects, such as diet, absorption,
`
`endogeny, and metabolism. Metabolism alone has various factors involved in regulation, such as changes
`in related enzyme activity, lipoproteins receptor expression, and gene transcription levels.
`
`Currently, it is known that high TG is an independent risk factor leading to As [7]. LPL and HP are
`
`two key enzymes related to endogenous TG metabolism in blood circulation, and they play an important
`role in catalyzing chylomicrons (CM) in plasma, very low-density lipoprotein (VLDL), and HDL metabolism.
`Defects or reduced activity can lead to blocked degradation of CM and VLDL, causing hypertriglyceridemia.
`LDL and HL activity is influenced by a variety of factors. This experiment found that PHLA, LPL, and HL
`activity was significantly lowered in the high-lipid model group. It could be that a high-lipid diet inhibits
`lipase synthesis by the liver and other organs [8]. Bisdemethoxycurcumin can cause a rise in PHLA, LPL, and
`HL activity, thereby speeding up the metabolism of TG and lipoproteins rich in TG and causing a notable
`drop in serum and liver TG content.
`
`The oxidative modification of lipoproteins has long been shown to be a key step leading to As [9],
`
`lipid peroxidation primarily is when oxygen free radicals produced by the body through enzyme systems
`or non-enzyme systems attack polyunsaturated fatty acids (PUFA) in the body, forming lipid peroxide.
`Therefore, measuring MDA can often reflect the degree of lipid peroxidation in the body, indirectly reflect
`the degree of cell damage, and predict the rate of As formation. Bisdemethoxycurcumin can significantly
`reduce MDA content, indicating that it has the effect of stopping As formation and easing As plaque that
`has already formed.
`
`With the rise in standards of living, the incidence and mortality rates of cardiovascular and
`
`cerebrovascular diseases resulting from hyperlipidemia have rapidly risen and tended toward younger
`ages, and there have been more and more cases of hypertriglyceridemia. For such diseases, current
`effective drug therapies in clinical settings primarily include statins, fibrates, niacin, and derivatives.
`However, because the question of whether these drugs have toxic side effects remains under debate, this
`has limited their application in clinical settings. The main source of bisdemethoxycurcumin is plant-based
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`turmeric. Turmeric is not only a traditional Chinese medicine, the curcumin it contains is a food coloring
`that has long been used by people, with no significant adverse reactions. The results of this experiment
`indicate that bisdemethoxycurcumin can reduce lipids, and its reduction of TG in particular is pronounced;
`also, it can fight lipid peroxidation and is expected to become an ideal lipid-lowering medicine.
`
`References
`
`[1] Wu Xingde, Gao Chengxian, Hong Xingqiu, et al. Experimental study of turmerol extract for diet-
`induced hyperlipidemia in rabbits [J] Journal of Zhejiang Chinese Medical University, 1999, 23 (1): 16 – 18.
`
`[2] Zhao Geping, Hong Xingqiu, Li Wanli, et al. Study of the effect of turmerol extract in fighting oxidative
`modification of LDL in vitro [J]. Journal of Zhejiang Chinese Medical University, 1999, 23 (2): 27 – 28.
`
`[3] Hong Xingqiu, Zhao Geping, Huang Yanfen, et al. Study of the influence of different components of
`turmerol extract on vascular smooth muscle cell proliferation [J]. Chin Arch Tradit Chin Med, 2004, 22 (10):
`1823-1824.
`
`[4] Li Jianming, Yang Heping, Liu Songqing. Experimental study of the effect of 3 types of curcumin
`monomers in inhibiting human endothelial cells [J]. Chongqing Med J, 2002, 31 (9): 804 – 805.
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`[5] Yang Mokun. Study of methods of measuring curcumin content [J]. Chongqing Med J, 1984, 13 (3): 49.
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`[6] Li Yunman, Lei Zhigang, Yang Guicheng. Comparison of the lipid-lowering effects of simvastatin and
`lovastatin [J]. China Pharm Univ, 2002, 33 (5): 448 – 450.
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`[7] Ye Ping. Blood lipids basics and clinical applications [M]. Beijing: People’s Military Medical Press, 2002.
`264.
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`[8] Wo Xingde, Cui Xiaoqiang, Tang Liuhua. Influence of curcumin on activity of enzymes associated with
`plasma lipoprotein metabolism [J]. Chin J Arterioscler, 2003, 11 (3): 223 – 226.
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